1 00:00:05,760 --> 00:00:07,520 WELCOME TO THE NIH DIRECTOR'S 2 00:00:07,520 --> 00:00:08,120 WEDNESDAY AFTERNOON LECTURE 3 00:00:08,120 --> 00:00:09,560 SERIES. 4 00:00:09,560 --> 00:00:15,680 I'M THE LEAD THE CELLULAR 5 00:00:15,680 --> 00:00:17,160 COMMUNICATION IN THE CHILD 6 00:00:17,160 --> 00:00:17,760 DEVELOPMENT INSTITUTE AND I 7 00:00:17,760 --> 00:00:17,960 AM 8 00:00:17,960 --> 00:00:20,360 THRILLED TO INTRODUCE DR. 9 00:00:20,360 --> 00:00:24,080 YISHI 10 00:00:24,080 --> 00:00:25,800 JIN, WHO IS CONDUCTING NEW 11 00:00:25,800 --> 00:00:30,800 RESEARCH IN NEURO- CELLULAR 12 00:00:30,800 --> 00:00:36,160 OUR SPEAKER TODAY IS YISHI 13 00:00:36,160 --> 00:00:36,400 JIN, A 14 00:00:36,400 --> 00:00:38,160 DISTINGUISHED PROF. OF 15 00:00:38,160 --> 00:00:39,120 NEUROBIOLOGY AND CELLULAR AND 16 00:00:39,120 --> 00:00:40,360 MOLECULAR MEDICINE AT THE 17 00:00:40,360 --> 00:00:41,520 UNIVERSITY OF CALIFORNIA SAN 18 00:00:41,520 --> 00:00:42,760 DIEGO. 19 00:00:42,760 --> 00:00:44,760 SHE HAS BEEN NOMINATED BY THE 20 00:00:44,760 --> 00:00:48,720 WOMEN SCIENTIST ADVISORS 21 00:00:48,720 --> 00:00:48,960 FIRST 22 00:00:48,960 --> 00:00:50,800 BECAUSE OF HER SCIENCE AND 23 00:00:50,800 --> 00:00:52,440 SECONDLY BECAUSE SHE MENTORED 24 00:00:52,440 --> 00:00:52,640 AND 25 00:00:52,640 --> 00:00:53,680 SERVED AS A ROLE MODEL FOR 26 00:00:53,680 --> 00:00:53,920 MANY 27 00:00:53,920 --> 00:00:56,160 WOMEN, INCLUDING MYSELF. 28 00:00:56,160 --> 00:00:58,960 DR. JIN RECEIVERS 29 00:00:58,960 --> 00:01:03,240 UNDERGRADUATE 30 00:01:03,240 --> 00:01:04,000 IN CELL BIOLOGY FROM PEKING 31 00:01:04,000 --> 00:01:04,640 UNIVERSITY AND COMPLETED HER 32 00:01:04,640 --> 00:01:04,840 PHD 33 00:01:04,840 --> 00:01:05,440 IN MOLECULAR BIOLOGY AT UC 34 00:01:05,440 --> 00:01:07,440 BERKELEY WORKING WITH -- 35 00:01:07,440 --> 00:01:07,720 MODULAR 36 00:01:07,720 --> 00:01:10,560 SYSTEM AT -- LABORATORY AND 37 00:01:10,560 --> 00:01:10,800 THEN 38 00:01:10,800 --> 00:01:12,240 SHE MOVED TO MIT FOR A 39 00:01:12,240 --> 00:01:12,520 POSTDOC 40 00:01:12,520 --> 00:01:14,360 WITH -- AND THERE IS WHERE 41 00:01:14,360 --> 00:01:14,560 SHE 42 00:01:14,560 --> 00:01:16,640 STARTED TO USE THE -- AS A 43 00:01:16,640 --> 00:01:16,880 MODEL 44 00:01:16,880 --> 00:01:17,960 SYSTEM. 45 00:01:17,960 --> 00:01:24,040 IN 1996, YIN STARTED HER LAB 46 00:01:24,040 --> 00:01:24,240 AT 47 00:01:24,240 --> 00:01:26,720 YOU SEE SANTA CRUZ AND JUST 48 00:01:26,720 --> 00:01:26,960 FIVE 49 00:01:26,960 --> 00:01:29,400 YEARS LATER SHE BECAME A 50 00:01:29,400 --> 00:01:29,680 HOWARD 51 00:01:29,680 --> 00:01:31,840 HUGHES INVESTIGATOR. 52 00:01:31,840 --> 00:01:32,880 TODAY SHE IS A DISTINGUISHED 53 00:01:32,880 --> 00:01:33,480 PROFESSOR AND CODIRECTOR OF 54 00:01:33,480 --> 00:01:37,720 THE 55 00:01:37,720 --> 00:01:40,760 -- INSTITUTE OF BRAIN AND 56 00:01:40,760 --> 00:01:40,960 MIND 57 00:01:40,960 --> 00:01:42,960 ACCUSES SAN DIEGO AND HER 58 00:01:42,960 --> 00:01:43,480 RESEARCH FOCUSES ON THE 59 00:01:43,480 --> 00:01:43,760 MOLECULAR 60 00:01:43,760 --> 00:01:45,960 GENETICS MECHANISM UNDERLYING 61 00:01:45,960 --> 00:01:46,560 DEVELOPMENT AND FUNCTIONING 62 00:01:46,560 --> 00:01:47,200 DEVELOPING SYSTEM AT FIRST HE 63 00:01:47,200 --> 00:01:52,160 PIONEERED THE USE OF 64 00:01:52,160 --> 00:01:52,800 FLUORESCENCE-- FOR INDIVIDUAL 65 00:01:52,800 --> 00:01:54,240 VISUALIZATION THE SYNOPTIC 66 00:01:54,240 --> 00:01:54,840 STRUCTURES AND THEN TO USE 67 00:01:54,840 --> 00:01:55,040 THE 68 00:01:55,040 --> 00:01:58,400 TOOL TO SCREEN FOR MOLECULES 69 00:01:58,400 --> 00:01:59,040 INVOLVED IN SYNAPTIC FAMILY 70 00:01:59,040 --> 00:01:59,240 AND 71 00:01:59,240 --> 00:01:59,880 DEVELOPMENT AND HER APPROACH 72 00:01:59,880 --> 00:02:03,080 PRACTICALLY ESTABLISHED THE 73 00:02:03,080 --> 00:02:04,920 FIELD 74 00:02:04,920 --> 00:02:05,560 OF MOLECULAR GENETICS AND IN 75 00:02:05,560 --> 00:02:07,760 ADDITION SHE PLAYED AND 76 00:02:07,760 --> 00:02:08,280 INSTRUMENTAL ROLE IN 77 00:02:08,280 --> 00:02:09,680 DEVELOPING 78 00:02:09,680 --> 00:02:10,880 THE LASER, TECHNOLOGY TO 79 00:02:10,880 --> 00:02:11,120 DEVELOP 80 00:02:11,120 --> 00:02:17,920 OUR MODERN RESEARCH ON OXY 81 00:02:17,920 --> 00:02:18,520 REGENERATION IN HER LAP NOT 82 00:02:18,520 --> 00:02:18,800 ONLY 83 00:02:18,800 --> 00:02:19,400 DISCOVERED THE MOLECULAR 84 00:02:19,400 --> 00:02:21,960 REASSEMBLY OR OXY 85 00:02:21,960 --> 00:02:22,320 REGENERATION 86 00:02:22,320 --> 00:02:26,040 AND ALSO -- DURING NERVOUS 87 00:02:26,040 --> 00:02:26,280 SYSTEM 88 00:02:26,280 --> 00:02:28,320 DEVELOPMENT. FOR HER SCIENCE 89 00:02:28,320 --> 00:02:28,520 YIN 90 00:02:28,520 --> 00:02:31,000 HAS RECEIVED NUMEROUS AWARDS 91 00:02:31,000 --> 00:02:32,360 FROM 92 00:02:32,360 --> 00:02:32,920 PRESIDENTIAL EARLY CAREER 93 00:02:32,920 --> 00:02:33,400 AWARD 94 00:02:33,400 --> 00:02:36,560 TO FELLOW OF AMERICAN SOCIETY 95 00:02:36,560 --> 00:02:38,800 OF 96 00:02:38,800 --> 00:02:39,440 ARTS AND SCIENCES; FELLOW OF 97 00:02:39,440 --> 00:02:39,680 THE 98 00:02:39,680 --> 00:02:40,520 AMERICAN SOCIETY OF CELL 99 00:02:40,520 --> 00:02:42,760 BIOLOGY, 100 00:02:42,760 --> 00:02:43,400 AND LAST MONTH MEMBER OF THE 101 00:02:43,400 --> 00:02:46,400 NATIONAL ACADEMY OF SCIENCES. 102 00:02:46,400 --> 00:02:47,760 YIN IS AMONG THE MOST HIGHLY 103 00:02:47,760 --> 00:02:48,880 CITED RESEARCHERS IN THE 104 00:02:48,880 --> 00:02:49,160 UNITED 105 00:02:49,160 --> 00:02:50,560 STATES. 106 00:02:50,560 --> 00:02:51,880 SHE IS ALSO DEDICATED TO 107 00:02:51,880 --> 00:02:54,560 MENTORSHIP AND SCIENCE 108 00:02:54,560 --> 00:02:54,880 EDUCATION; 109 00:02:54,880 --> 00:02:56,120 SHE HAS TRAINED DOZENS OF 110 00:02:56,120 --> 00:02:56,360 YOUNG 111 00:02:56,360 --> 00:02:57,840 SCIENTISTS MANY OF WHOM ARE 112 00:02:57,840 --> 00:02:58,800 NOW 113 00:02:58,800 --> 00:03:01,720 IN TENURE OR TENURE-TRACK 114 00:03:01,720 --> 00:03:03,000 POSITIONS ALL OVER THE WORLD 115 00:03:03,000 --> 00:03:03,200 AND 116 00:03:03,200 --> 00:03:05,040 THE TITLE OF HER TALK TODAY 117 00:03:05,040 --> 00:03:05,240 IS 118 00:03:05,240 --> 00:03:06,680 NEURONAL RESPONSE TO INJURY 119 00:03:06,680 --> 00:03:06,880 AND 120 00:03:06,880 --> 00:03:11,880 STRESS INSIDE -- ELEGANCE. 121 00:03:11,880 --> 00:03:12,560 WELCOME TODAY. 122 00:03:12,560 --> 00:03:15,280 THANK YOU VERY MUCH FOR THAT 123 00:03:15,280 --> 00:03:15,520 VERY 124 00:03:15,520 --> 00:03:20,200 GENEROUS INTRODUCTION, AND 125 00:03:20,200 --> 00:03:20,440 ALSO 126 00:03:20,440 --> 00:03:22,840 THANK YOU FOR NOMINATING ME 127 00:03:22,840 --> 00:03:23,040 FOR 128 00:03:23,040 --> 00:03:24,560 THIS LECTURE AND IT IS REALLY 129 00:03:24,560 --> 00:03:26,520 HUMBLING TO SPEAK HERE, 130 00:03:26,520 --> 00:03:27,120 PARTICULARLY APPRECIATE ALL 131 00:03:27,120 --> 00:03:29,440 THE 132 00:03:29,440 --> 00:03:35,440 SUPPORT OF THE NIH TO MY 133 00:03:35,440 --> 00:03:36,720 PROGRAM 134 00:03:36,720 --> 00:03:38,560 AND AS THE TITLE IMPLIES. I 135 00:03:38,560 --> 00:03:39,240 WILL 136 00:03:39,240 --> 00:03:41,920 COVER FROM THE WORK WE USE, C 137 00:03:41,920 --> 00:03:44,760 ELEGANCE AS A MODEL TO 138 00:03:44,760 --> 00:03:45,080 UNDERSTAND 139 00:03:45,080 --> 00:03:46,920 THE FUNDAMENTAL COMMUNITY 140 00:03:46,920 --> 00:03:47,160 SOME 141 00:03:47,160 --> 00:03:50,880 HOW NEURONS PROTECT 142 00:03:50,880 --> 00:03:51,200 THEMSELVES 143 00:03:51,200 --> 00:03:53,720 THAT I WILL START WITH THIS 144 00:03:53,720 --> 00:03:55,320 CENTURY-OLD IMAGE, QUITE 145 00:03:55,320 --> 00:03:55,560 PLENTY 146 00:03:55,560 --> 00:04:00,760 OF THEM AT UC AT THE NH 147 00:04:00,760 --> 00:04:01,760 CAMPUS 148 00:04:01,760 --> 00:04:07,400 PRODUCED BY RAMON Y CAJAL. 149 00:04:07,400 --> 00:04:09,320 AND THROUGH VERY SYSTEMATIC 150 00:04:09,320 --> 00:04:09,520 AND 151 00:04:09,520 --> 00:04:10,000 EXTENSIVE MICROSCOPY 152 00:04:10,000 --> 00:04:12,200 OBSERVATIONS 153 00:04:12,200 --> 00:04:12,840 -- I AM TRYING TO FIGURE OUT 154 00:04:12,840 --> 00:04:13,040 IF 155 00:04:13,040 --> 00:04:23,400 THIS IS THE LASER. 156 00:04:23,680 --> 00:04:25,920 I'M SUPPOSED TO BE USING THE 157 00:04:25,920 --> 00:04:27,720 LASER ON MY SCREEN? 158 00:04:27,720 --> 00:04:30,760 I THINK YOU CAN SEE THAT. 159 00:04:30,760 --> 00:04:32,360 THAT IS THROUGH A LOT OF WORK 160 00:04:32,360 --> 00:04:33,720 HIMSELF AND HIS STUDENTS, 161 00:04:33,720 --> 00:04:37,800 AND THEY DEMONSTRATED NOT 162 00:04:37,800 --> 00:04:38,040 ONLY 163 00:04:38,040 --> 00:04:42,840 THE NEURON -- AND LATER 164 00:04:42,840 --> 00:04:43,160 EXPLORED 165 00:04:43,160 --> 00:04:46,360 THE CONCEPT OF HOW NEURONS, 166 00:04:46,360 --> 00:04:46,560 HOW 167 00:04:46,560 --> 00:04:48,240 DID THEY RESPOND TO INJURY. 168 00:04:48,240 --> 00:04:52,080 AND THIS IMAGE ACTUALLY SHOWS 169 00:04:52,080 --> 00:04:52,280 THE 170 00:04:52,280 --> 00:04:54,000 DALCAMA (PHONETIC) IN THE 171 00:04:54,000 --> 00:04:54,920 CENTRAL 172 00:04:54,920 --> 00:05:01,400 NERVOUS SYSTEM RESPONDING TO 173 00:05:01,400 --> 00:05:01,560 A 174 00:05:01,560 --> 00:05:02,960 KNIFE CUT AND THE EXONS, 175 00:05:02,960 --> 00:05:12,800 LOOKING AT A SIGN OF THE 176 00:05:12,800 --> 00:05:16,840 DISTROPHIC BULB, 177 00:05:16,840 --> 00:05:20,680 WHICH TRIGGERED A LOT OF 178 00:05:20,680 --> 00:05:21,000 STUDIES 179 00:05:21,000 --> 00:05:21,640 AND SOME CONTRIBUTED TO THE 180 00:05:21,640 --> 00:05:22,520 ENVIRONMENT, 181 00:05:22,520 --> 00:05:26,080 AND SOME CONTRIBUTED TO 182 00:05:26,080 --> 00:05:28,680 INTRACELLULAR DISORGANIZATION 183 00:05:28,680 --> 00:05:28,880 OF 184 00:05:28,880 --> 00:05:29,480 CYTOSKELETON SUCH AS THE 185 00:05:29,480 --> 00:05:29,680 STUDY 186 00:05:29,680 --> 00:05:37,760 DONE BY FRANK BRATSBUG'S LAB 187 00:05:37,760 --> 00:05:39,840 (PHONETIC). 188 00:05:39,840 --> 00:05:40,960 AND IT HAS EVOLVED OVER THE 189 00:05:40,960 --> 00:05:42,400 YEARS 190 00:05:42,400 --> 00:05:50,680 BOTH TO INHIBITING GULLIAR 191 00:05:50,680 --> 00:05:50,960 SCALLS 192 00:05:50,960 --> 00:05:51,560 INHIBITORS AND ALSO MARKER 193 00:05:51,560 --> 00:05:54,280 TUBULAR -- AND TO PROTECT THE 194 00:05:54,280 --> 00:05:56,040 -- 195 00:05:56,040 --> 00:05:59,680 TO REGENERATING. 196 00:05:59,680 --> 00:06:00,280 DESPITE OF THESE MANY YEARS 197 00:06:00,280 --> 00:06:02,120 EFFORTS OUR UNDERSTANDING OF 198 00:06:02,120 --> 00:06:06,640 HOW 199 00:06:06,640 --> 00:06:07,200 NEURONS RESPOND TO INJURY 200 00:06:07,200 --> 00:06:07,480 REMAINS 201 00:06:07,480 --> 00:06:09,600 AN INTENSE TOPIC WITH MANY 202 00:06:09,600 --> 00:06:13,520 THEORIES. 203 00:06:13,520 --> 00:06:14,120 I HAVE BEEN WORKING WITH A 204 00:06:14,120 --> 00:06:14,360 MODEL 205 00:06:14,360 --> 00:06:18,920 ORGANISM FOR 20-30 YEARS 206 00:06:18,920 --> 00:06:24,720 PARTICULAR USING NUMATOAD ODS 207 00:06:24,720 --> 00:06:26,280 25TH ANNIVERSARY SCIENTIFIC 208 00:06:26,280 --> 00:06:29,080 SYMPOSIUM HERE. 209 00:06:29,080 --> 00:06:32,000 (CORRECTION) 210 00:06:32,000 --> 00:06:40,920 ODS 25TH ANNIVERSARY -- 211 00:06:40,920 --> 00:06:42,080 (CORRECTION) 212 00:06:42,080 --> 00:06:43,920 C ELEGANS. 213 00:06:43,920 --> 00:06:49,800 IN C ELEGANS AXONS GROW 214 00:06:49,800 --> 00:06:50,120 AROUND A 215 00:06:50,120 --> 00:06:52,200 MATRIX AND ARE NOT 216 00:06:52,200 --> 00:06:54,080 MYELINATED, 217 00:06:54,080 --> 00:06:54,920 GENERALLY THEIR MORPHOLOGY IS 218 00:06:54,920 --> 00:07:01,880 RATHER SIMPLE. IN MANY 219 00:07:01,880 --> 00:07:02,240 STUDIES IN 220 00:07:02,240 --> 00:07:04,320 SERENDIPITOUS WAYS WE APPLY 221 00:07:04,320 --> 00:07:06,280 THE 222 00:07:06,280 --> 00:07:07,800 PHYSICS, AND IT WAS A WAY TO 223 00:07:07,800 --> 00:07:11,280 SEVER THE C ELEGANS AXON. 224 00:07:11,280 --> 00:07:12,960 THIS SHOWS A MOVING WHAT YOU 225 00:07:12,960 --> 00:07:13,160 SEE 226 00:07:13,160 --> 00:07:21,560 THIS IS A LASER CUT AND THIS 227 00:07:21,560 --> 00:07:21,880 IS 228 00:07:21,880 --> 00:07:24,520 (INDISCERNIBLE) -- 229 00:07:24,520 --> 00:07:25,840 AND THIS BEGINS TO INITIATE 230 00:07:25,840 --> 00:07:26,200 GROSS 231 00:07:26,200 --> 00:07:31,600 CONES AND VERY SPONTANEOUS, 232 00:07:31,600 --> 00:07:31,840 VERY 233 00:07:31,840 --> 00:07:33,240 RAPID BUT THIS REGROWTH IS 234 00:07:33,240 --> 00:07:35,960 ACTUALLY VERY SPONTANEOUS. 235 00:07:35,960 --> 00:07:37,560 DOES NOT ACTUALLY REPAIR 236 00:07:37,560 --> 00:07:37,840 ITSELF 237 00:07:37,840 --> 00:07:40,640 AND CAN PULL BACK TO ITS 238 00:07:40,640 --> 00:07:42,720 DEVELOPMENTAL STATE. 239 00:07:42,720 --> 00:07:45,920 SO WHEN WE ESTABLISHED LASER 240 00:07:45,920 --> 00:07:49,000 AXOLTOMY (PHONETIC) ASSAY, 241 00:07:49,000 --> 00:07:49,840 WE DECIDED TO TAKE ADVANTAGE 242 00:07:49,840 --> 00:07:51,560 OF C 243 00:07:51,560 --> 00:07:54,280 ELEGANS SINGLE CELL AND 244 00:07:54,280 --> 00:07:54,560 DECIDED 245 00:07:54,560 --> 00:07:58,360 TO PROBE INTO THE UNDERLYING 246 00:07:58,360 --> 00:08:00,040 MECHANISM. 247 00:08:00,040 --> 00:08:03,400 THE FIRST THING IS WE KNOW, 248 00:08:03,400 --> 00:08:04,760 GENERALLY CALCIUM IS A FIRST 249 00:08:04,760 --> 00:08:06,000 RESPONDER TO A LOT OF 250 00:08:06,000 --> 00:08:06,280 TRAUMATIC 251 00:08:06,280 --> 00:08:09,400 INJURIES. SO WE USED IN THESE 252 00:08:09,400 --> 00:08:10,040 EARLY STUDIES ABOUT A DECADE 253 00:08:10,040 --> 00:08:10,360 AGO, 254 00:08:10,360 --> 00:08:20,920 WE EXPRESSED THE G COM AND C 255 00:08:22,560 --> 00:08:23,080 ELEGANS NEURONS AS YOU CAN 256 00:08:23,080 --> 00:08:23,280 SEE 257 00:08:23,280 --> 00:08:28,280 HERE FOLLOWING SURGERY 258 00:08:28,280 --> 00:08:30,920 AND YOU CAN SEE THE TRACE OF 259 00:08:30,920 --> 00:08:31,480 CALCIUM AND ORANGE COLOR. 260 00:08:31,480 --> 00:08:32,360 THIS IS ALSO VERY SPONTANEOUS 261 00:08:32,360 --> 00:08:32,560 BUT 262 00:08:32,560 --> 00:08:34,600 WE CAN MEASURE THE TRANSIENT 263 00:08:34,600 --> 00:08:37,360 AMPLITUDE OF CUT SITE TO 264 00:08:37,360 --> 00:08:38,920 TOTAL 265 00:08:38,920 --> 00:08:41,880 GROWTH EXTEND WHERE WE CAN 266 00:08:41,880 --> 00:08:42,120 SEE A 267 00:08:42,120 --> 00:08:43,240 POSITIVE CORRELATION, 268 00:08:43,240 --> 00:08:47,480 AND SUGGESTING THE CALCIUM 269 00:08:47,480 --> 00:08:48,040 TRANSIENT COMES WITH THE 270 00:08:48,040 --> 00:08:48,280 INJURY 271 00:08:48,280 --> 00:08:52,920 LIKELY CONTRIBUTING TO THE 272 00:08:52,920 --> 00:08:56,880 INJURED AXON TO REGROW AND 273 00:08:56,880 --> 00:08:57,080 WE 274 00:08:57,080 --> 00:09:00,440 WERE ABLE TO LINK THE CALCIUM 275 00:09:00,440 --> 00:09:03,600 TRANSIENT TO BULK -- CALCIUM 276 00:09:03,600 --> 00:09:07,160 CHANNEL, AND THIS CALCIUM 277 00:09:07,160 --> 00:09:08,600 TRANSIENT WORKS TOGETHER WITH 278 00:09:08,600 --> 00:09:09,440 SECOND MESSENGER SYSTEMS TO 279 00:09:09,440 --> 00:09:13,920 PROMOTE AXON REGROWTH. 280 00:09:13,920 --> 00:09:19,160 IN MANY WAYS THAT PEACE 281 00:09:19,160 --> 00:09:19,480 PROMOTES 282 00:09:19,480 --> 00:09:21,240 THE PERIPHERAL NERVE TO 283 00:09:21,240 --> 00:09:22,400 REGENERATE AND IT HAS LONG 284 00:09:22,400 --> 00:09:22,640 BEEN 285 00:09:22,640 --> 00:09:23,440 STUDIED IN THE MAMMALIAN 286 00:09:23,440 --> 00:09:24,120 FIELD 287 00:09:24,120 --> 00:09:29,080 PARTICULARLY IN WOLVES AND IN 288 00:09:29,080 --> 00:09:29,680 (INDISCERNIBLE) MANY YEARS 289 00:09:29,680 --> 00:09:30,640 AGO. 290 00:09:30,640 --> 00:09:33,280 MORE RECENTLY WE IDENTIFIED 291 00:09:33,280 --> 00:09:36,240 THIS 292 00:09:36,240 --> 00:09:40,960 RAPID AXON-CALCIUM TRANSIENT 293 00:09:40,960 --> 00:09:42,920 FOLLOWED BY A SLOWER CALCIUM 294 00:09:42,920 --> 00:09:44,640 TRANSIENT IN MITOCHONDRIA 295 00:09:44,640 --> 00:09:44,880 WHICH 296 00:09:44,880 --> 00:09:52,120 DEPENDS ON MITO UNI POLAR, 297 00:09:52,120 --> 00:09:52,520 (INDISCERNIBLE). 298 00:09:52,520 --> 00:09:53,960 --- 299 00:09:53,960 --> 00:09:55,960 ADDITIONALLY WE HAVE BEEN 300 00:09:55,960 --> 00:10:00,240 EXPLORING BOTH MEMBRANE 301 00:10:00,240 --> 00:10:02,480 ORGANELLES AS WELL AS 302 00:10:02,480 --> 00:10:03,080 CYTOSKELETON, AND ONE OTHER 303 00:10:03,080 --> 00:10:03,320 VERY 304 00:10:03,320 --> 00:10:06,720 TRADITIONAL MARKERS LOOKING 305 00:10:06,720 --> 00:10:06,920 AT 306 00:10:06,920 --> 00:10:12,600 CYTOSKELETON AS A MICROTUBULE 307 00:10:12,600 --> 00:10:12,800 308 00:10:12,800 --> 00:10:13,800 PROCESS BITING FACTOR CALLED 309 00:10:13,800 --> 00:10:14,000 THE 310 00:10:14,000 --> 00:10:14,800 EBP. 311 00:10:14,800 --> 00:10:16,400 YOU SEE A SINGLE AXON, 312 00:10:16,400 --> 00:10:20,040 PROBABLY ONLY 10-15 313 00:10:20,040 --> 00:10:20,400 MICROTUBULE 314 00:10:20,400 --> 00:10:22,520 FIBERS. 315 00:10:22,520 --> 00:10:23,120 IN THIS MOVIE YOU WILL SEE 316 00:10:23,120 --> 00:10:23,400 INJURY 317 00:10:23,400 --> 00:10:30,920 SITE AND THE EBP:GFP FORMS 318 00:10:30,920 --> 00:10:31,160 WHITE 319 00:10:31,160 --> 00:10:32,960 SPOTS WHICH CONVERT TO THE 320 00:10:32,960 --> 00:10:41,840 CHAMOGRAPH AND THE EBP SITES 321 00:10:41,840 --> 00:10:42,320 CORRELATING TO THE 322 00:10:42,320 --> 00:10:42,920 MICROTUBULE 323 00:10:42,920 --> 00:10:45,760 DYNAMICS. 324 00:10:45,760 --> 00:10:49,000 INJURED AXONS HARDLY SEE EBP 325 00:10:49,000 --> 00:10:55,880 COME 326 00:10:55,880 --> 00:10:56,720 IN AND WITHIN THREE HOURS WE 327 00:10:56,720 --> 00:10:57,240 START TO SEE SIGNS OF 328 00:10:57,240 --> 00:10:57,560 PHILIPODIA 329 00:10:57,560 --> 00:10:58,720 (PHONETIC) 330 00:10:58,720 --> 00:10:59,240 AND WE START TO SEE AN 331 00:10:59,240 --> 00:10:59,560 INCREASE 332 00:10:59,560 --> 00:11:07,960 IN EBP CLASS NUMBERS, IT 333 00:11:07,960 --> 00:11:08,240 BEGINS 334 00:11:08,240 --> 00:11:09,160 TO (INDISCERNIBLE) AND WE 335 00:11:09,160 --> 00:11:09,920 START 336 00:11:09,920 --> 00:11:18,400 TO SEE THE EBP COME IN, FIRST 337 00:11:18,400 --> 00:11:19,560 INCREASE AND THEN SUSTAIN THE 338 00:11:19,560 --> 00:11:24,440 GROWTH IN RUNNING LANS. 339 00:11:24,440 --> 00:11:25,080 WE IDENTIFIED A MOLECULE 340 00:11:25,080 --> 00:11:25,360 CALLED 341 00:11:25,360 --> 00:11:35,120 THE -- A MICROTUBULE 342 00:11:35,120 --> 00:11:36,400 STABILIZING 343 00:11:36,400 --> 00:11:37,280 FACTOR, AND THAT TRIGGERS A 344 00:11:37,280 --> 00:11:38,800 DESTABILIZING (INDISCERNIBLE) 345 00:11:38,800 --> 00:11:39,000 TO 346 00:11:39,000 --> 00:11:41,480 WORK TOGETHER WITH TO 347 00:11:41,480 --> 00:11:41,760 CONSERVE 348 00:11:41,760 --> 00:11:45,480 THE PROTEIN CALLED THE TACC 349 00:11:45,480 --> 00:11:45,680 AND 350 00:11:45,680 --> 00:11:47,160 (INDISCERNIBLE). 351 00:11:47,160 --> 00:11:48,320 THIS FUNDING SUBSEQUENTLY 352 00:11:48,320 --> 00:11:52,240 FOLLOWED BY JAMES FOCET'S 353 00:11:52,240 --> 00:11:58,920 (PHONETIC) GROUP, SHOWS 354 00:11:58,920 --> 00:11:59,200 PERIPHERY 355 00:11:59,200 --> 00:12:03,480 NERVE MAMMALIAN --. 356 00:12:03,480 --> 00:12:08,480 --- 357 00:12:08,480 --> 00:12:11,400 I'M GOING TO SET UP THIS 358 00:12:11,400 --> 00:12:12,720 LASER 359 00:12:12,720 --> 00:12:18,960 EXOLTAMY ASSAY TO TAKE AGING 360 00:12:18,960 --> 00:12:19,560 CONSERVATION SCREEN TO LOOK 361 00:12:19,560 --> 00:12:21,040 FOR A 362 00:12:21,040 --> 00:12:21,600 MECHANISM THAT COULD BE 363 00:12:21,600 --> 00:12:26,600 SHARED 364 00:12:26,600 --> 00:12:27,200 -- LOOKING AT THE MAMMALIAN 365 00:12:27,200 --> 00:12:27,400 AXON 366 00:12:27,400 --> 00:12:29,040 REGENERATION FIELD. THE WAY 367 00:12:29,040 --> 00:12:29,240 WE 368 00:12:29,240 --> 00:12:36,280 DID IT IS THAT C ELEGANS 369 00:12:36,280 --> 00:12:41,800 GENERALLY IS IN CODING GENES. 370 00:12:41,800 --> 00:12:43,320 WE HAVE WONDERFUL RESOURCE 371 00:12:43,320 --> 00:12:45,600 COMMUNITY THAT GENERATED LOTS 372 00:12:45,600 --> 00:12:45,800 OF 373 00:12:45,800 --> 00:12:47,920 GENETIC MUTATIONS, AND SO WE 374 00:12:47,920 --> 00:12:48,120 THEN 375 00:12:48,120 --> 00:12:48,760 STARTED CHOOSING ABOUT 1500 376 00:12:48,760 --> 00:12:49,240 OF 377 00:12:49,240 --> 00:12:54,160 THEM THAT WE KNOW COULD BE 378 00:12:54,160 --> 00:12:55,920 EXPRESSED IN THE NERVOUS 379 00:12:55,920 --> 00:12:56,160 SYSTEM 380 00:12:56,160 --> 00:12:57,520 AND COULD BE INFLUENCING AXON 381 00:12:57,520 --> 00:13:00,040 REGENERATION. 382 00:13:00,040 --> 00:13:02,400 IN ALL THESE CASES WE CHOOSE 383 00:13:02,400 --> 00:13:02,760 THE 384 00:13:02,760 --> 00:13:06,640 GENES -- GENERALLY WITHIN 385 00:13:06,640 --> 00:13:09,080 NORMAL 386 00:13:09,080 --> 00:13:13,040 NEURO SYSTEM ARCHITECTURE AND 387 00:13:13,040 --> 00:13:13,800 PERFORMED IT IN LATE LARVAE 388 00:13:13,800 --> 00:13:14,040 ADULT 389 00:13:14,040 --> 00:13:15,040 STAGE LOOKING FOR AXON 390 00:13:15,040 --> 00:13:18,600 REGENERATION RESPONSE, MORE 391 00:13:18,600 --> 00:13:19,160 RESEMBLING THE MAMMALIAN 392 00:13:19,160 --> 00:13:19,640 REGENERATION FIELD. 393 00:13:19,640 --> 00:13:20,400 --- 394 00:13:20,400 --> 00:13:22,160 AS SHOWN HERE, 395 00:13:22,160 --> 00:13:24,800 ESSENTIALLY WE SCREENED ALL 396 00:13:24,800 --> 00:13:25,000 OF 397 00:13:25,000 --> 00:13:27,680 THOSE 1500 GENES; THE 398 00:13:27,680 --> 00:13:28,360 MAJORITY OF 399 00:13:28,360 --> 00:13:32,400 THEM ALMOST 80% OF THEM HAD 400 00:13:32,400 --> 00:13:32,600 NO 401 00:13:32,600 --> 00:13:33,320 DEFECT AND WHAT I MEAN BY 402 00:13:33,320 --> 00:13:33,760 THAT IS 403 00:13:33,760 --> 00:13:36,000 SHOWN IN THIS IMAGE. 404 00:13:36,000 --> 00:13:37,880 IN RESPONDING TO INJURY, AND 405 00:13:37,880 --> 00:13:38,080 THE 406 00:13:38,080 --> 00:13:43,040 RED ARROW, THOSE AXONS WILL 407 00:13:43,040 --> 00:13:44,040 INITIALLY REGROW BUT YOU CAN 408 00:13:44,040 --> 00:13:44,240 SEE 409 00:13:44,240 --> 00:13:48,920 THAT IT'S GROWTH IS RANDOM, 410 00:13:48,920 --> 00:13:49,120 ALL 411 00:13:49,120 --> 00:13:51,680 OVER THE PLACE AND THIS 412 00:13:51,680 --> 00:13:52,000 REGROWTH 413 00:13:52,000 --> 00:13:52,560 REPRESENTS A SPONTANEOUS 414 00:13:52,560 --> 00:13:52,880 RESPONSE 415 00:13:52,880 --> 00:13:53,200 TO INJURY. 416 00:13:53,200 --> 00:13:56,040 WE SEE ABOUT 15% OF THE GENES 417 00:13:56,040 --> 00:14:05,160 THAT HAVE LOSS OF FUNCTIONING 418 00:14:05,160 --> 00:14:06,400 IN 419 00:14:06,400 --> 00:14:07,000 THOSE GENES, RESULTING IN A 420 00:14:07,000 --> 00:14:08,760 DECREASE IN THE REGROWTH TO 421 00:14:08,760 --> 00:14:11,240 PROMOTE INJURY RESPONSE AND 422 00:14:11,240 --> 00:14:11,480 THERE 423 00:14:11,480 --> 00:14:12,120 IS NOT MUCH REGROWTH IN THIS 424 00:14:12,120 --> 00:14:12,800 IMAGE VERY LITTLE BRANCHING. 425 00:14:12,800 --> 00:14:14,640 --- 426 00:14:14,640 --> 00:14:20,160 ABOUT 5% OF THE GENES ACT AS 427 00:14:20,160 --> 00:14:22,280 AXON 428 00:14:22,280 --> 00:14:22,600 INHIBITORS. 429 00:14:22,600 --> 00:14:23,160 THE TOTAL GROWTH IS MORE 430 00:14:23,160 --> 00:14:23,480 COMPARED 431 00:14:23,480 --> 00:14:25,720 TO (INDISCERNIBLE). 432 00:14:25,720 --> 00:14:33,160 BECAUSE THESE GENES ARE 433 00:14:33,160 --> 00:14:33,400 CHOSEN 434 00:14:33,400 --> 00:14:35,160 BASED ON HOMOLOGY -- YOU CAN 435 00:14:35,160 --> 00:14:35,360 SEE 436 00:14:35,360 --> 00:14:39,000 THIS IN HUMANS FROM GENE 437 00:14:39,000 --> 00:14:42,400 EXPRESSION TO -- AND THIS 438 00:14:42,400 --> 00:14:42,640 SUMMARY 439 00:14:42,640 --> 00:14:48,720 SHOWS THAT USING EACH GENE 440 00:14:48,720 --> 00:14:56,800 GOAL 441 00:14:56,800 --> 00:14:57,360 CATEGORIES, THERE WILL BE 442 00:14:57,360 --> 00:14:57,600 GENES 443 00:14:57,600 --> 00:14:58,200 REQUIRED FOR PROMOTING THE 444 00:14:58,200 --> 00:14:59,800 INJURED ACCENT TO GROW, 445 00:14:59,800 --> 00:15:05,000 AND GENES ACTING TO PROMOTE 446 00:15:05,000 --> 00:15:05,200 THE 447 00:15:05,200 --> 00:15:06,560 REGROWTH, AND AS A 448 00:15:06,560 --> 00:15:08,560 GENETICIST, 449 00:15:08,560 --> 00:15:11,520 AXON REGENERATION FOLLOWING 450 00:15:11,520 --> 00:15:12,800 INJURY IS A PROCESS THAT 451 00:15:12,800 --> 00:15:13,080 SOMEHOW 452 00:15:13,080 --> 00:15:13,600 MIMICS A DEVELOPMENTAL 453 00:15:13,600 --> 00:15:19,840 FORMING OF 454 00:15:19,840 --> 00:15:20,880 THE WHOLE NERVOUS SYSTEM, 455 00:15:20,880 --> 00:15:23,080 REQUIRING (INDISCERNIBLE) OF 456 00:15:23,080 --> 00:15:23,240 A 457 00:15:23,240 --> 00:15:27,000 VARIETY OF GENES. WE OFTEN 458 00:15:27,000 --> 00:15:27,200 SAY 459 00:15:27,200 --> 00:15:27,760 THERE IS A SINGLE MASTER 460 00:15:27,760 --> 00:15:36,720 REGULATOR TO CONTROL AXON 461 00:15:36,720 --> 00:15:37,320 REGENERATION WHETHER YOU 462 00:15:37,320 --> 00:15:37,560 PREPARE 463 00:15:37,560 --> 00:15:38,000 YOURSELF FOR NOT. 464 00:15:38,000 --> 00:15:46,080 --- 465 00:15:46,080 --> 00:15:47,320 TAKING A MOLECULAR GENETICIST 466 00:15:47,320 --> 00:15:47,760 APPROACH, THIS IS 467 00:15:47,760 --> 00:15:48,120 CATEGORIZING 468 00:15:48,120 --> 00:15:52,080 THE KEY GENES, EXPRESSING 469 00:15:52,080 --> 00:15:56,360 THOSE 470 00:15:56,360 --> 00:15:57,920 GENE MANIPULATING GENE 471 00:15:57,920 --> 00:15:58,240 FUNCTION 472 00:15:58,240 --> 00:15:58,760 AND ENABLE IMAGING, AND 473 00:15:58,760 --> 00:15:59,080 GENERALLY 474 00:15:59,080 --> 00:16:02,920 WE DIVIDE THE AXON INJURY 475 00:16:02,920 --> 00:16:05,400 RESPONSE INTO THREE STAGES. 476 00:16:05,400 --> 00:16:06,920 THE VERY IMMEDIATE STAGES A 477 00:16:06,920 --> 00:16:10,680 CALCIUM TRANSIENT WHICH THEN 478 00:16:10,680 --> 00:16:10,880 IS 479 00:16:10,880 --> 00:16:11,480 FOLLOWED BY THE INITIATION 480 00:16:11,480 --> 00:16:12,680 PHASE 481 00:16:12,680 --> 00:16:22,160 OF REGROWTH, MEMBRANE REPAIR, 482 00:16:22,160 --> 00:16:23,000 REORGANIZATION FOLLOWING THE 483 00:16:23,000 --> 00:16:27,600 EXTENSION PHASE WHERE IT CAN 484 00:16:27,600 --> 00:16:27,840 GROW 485 00:16:27,840 --> 00:16:29,360 MORE OR LESS IN WHAT WE FIND 486 00:16:29,360 --> 00:16:29,560 IS 487 00:16:29,560 --> 00:16:33,680 OUR FOCUS TODAY MOSTLY IS ON 488 00:16:33,680 --> 00:16:34,920 THE 489 00:16:34,920 --> 00:16:35,760 DLK-KINASE. 490 00:16:35,760 --> 00:16:42,520 AND A BIT ABOUT THE PROTEIN- 491 00:16:42,520 --> 00:16:46,800 FINDING, PHASE SEPARATION AND 492 00:16:46,800 --> 00:16:51,080 LATER I WILL TALK ABOUT 493 00:16:51,080 --> 00:16:54,400 TRANSLATING CELF FUNDING INTO 494 00:16:54,400 --> 00:16:58,840 MAMMALS AND COME BACK TO -- 495 00:16:58,840 --> 00:17:00,480 INVOLVING REGULATION AND IS 496 00:17:00,480 --> 00:17:00,680 IN 497 00:17:00,680 --> 00:17:06,920 THE LATEST -- ELONGATION OF 498 00:17:06,920 --> 00:17:07,160 THE 499 00:17:07,160 --> 00:17:07,680 DEVELOPMENTAL GUIDANCE 500 00:17:07,680 --> 00:17:08,000 MOLECULES 501 00:17:08,000 --> 00:17:09,720 START COMING TO PLAY TO 502 00:17:09,720 --> 00:17:10,040 MODULATE 503 00:17:10,040 --> 00:17:13,440 THE GROWTH DIRECTION AND 504 00:17:13,440 --> 00:17:13,720 GROWTH 505 00:17:13,720 --> 00:17:13,960 EXTENT. 506 00:17:13,960 --> 00:17:14,920 --- 507 00:17:14,920 --> 00:17:15,480 LET ME START WITH THIS 508 00:17:15,480 --> 00:17:15,720 PROTEIN 509 00:17:15,720 --> 00:17:22,920 CALLED THE TIAR-2, AN 510 00:17:22,920 --> 00:17:24,360 RI-BINDING 511 00:17:24,360 --> 00:17:25,440 FACTOR, EXTENDING IT WITH 512 00:17:25,440 --> 00:17:26,600 THREE 513 00:17:26,600 --> 00:17:29,840 BINDING DOMAINS. 514 00:17:29,840 --> 00:17:34,840 IT'S NAMED AS A TIA BECAUSE 515 00:17:34,840 --> 00:17:35,080 IT IS 516 00:17:35,080 --> 00:17:38,920 A PROTEIN EXPRESSED, A T-CELL 517 00:17:38,920 --> 00:17:39,120 AS 518 00:17:39,120 --> 00:17:41,400 AN ANTIGEN. 519 00:17:41,400 --> 00:17:42,040 AND WHAT CHARACTERIZES THIS 520 00:17:42,040 --> 00:17:45,080 FAMILY IS BESIDES THE THREE 521 00:17:45,080 --> 00:17:49,840 DOMAINS, IT HAS A PRION-LIKE 522 00:17:49,840 --> 00:17:54,000 DOMAIN. 523 00:17:54,000 --> 00:17:54,600 IF YOU YEARS AGO THERE WERE 524 00:17:54,600 --> 00:17:55,960 REPORTS ABOUT 525 00:17:55,960 --> 00:17:59,440 MUTATIONS IN THE HUMAN TIA 526 00:17:59,440 --> 00:18:00,800 GENES 527 00:18:00,800 --> 00:18:02,800 ACTUALLY LINK TO STRESS-- 528 00:18:02,800 --> 00:18:06,920 DYNAMICS AS WELL AS ALS 529 00:18:06,920 --> 00:18:07,360 PATIENTS. 530 00:18:07,360 --> 00:18:10,920 --- 531 00:18:10,920 --> 00:18:15,720 A FORMER POST-DOC 532 00:18:15,720 --> 00:18:16,120 CHARACTERIZED A 533 00:18:16,120 --> 00:18:19,520 LOT OF BINDING FACTORS, LOTS 534 00:18:19,520 --> 00:18:22,120 OF 535 00:18:22,120 --> 00:18:22,720 (INDISCERNIBLE) WITH MORE 536 00:18:22,720 --> 00:18:24,520 EXTENDED GROWTH AFTER INJURY 537 00:18:24,520 --> 00:18:28,440 (TIAR-2), 538 00:18:28,440 --> 00:18:30,840 AND WE WILL NORMALIZE THIS 539 00:18:30,840 --> 00:18:34,360 GROWTH, ANYTHING ABOVE ONE IT 540 00:18:34,360 --> 00:18:35,280 SHOWS THAT LOSING TIAR-2 541 00:18:35,280 --> 00:18:37,560 LEADS TO 542 00:18:37,560 --> 00:18:40,640 MORE REGROWTH EXTENSION. 543 00:18:40,640 --> 00:18:47,000 AND IT SHOWS TIAR-2 ACTING IN 544 00:18:47,000 --> 00:18:47,200 THE 545 00:18:47,200 --> 00:18:48,200 EARLY PHASE. 546 00:18:48,200 --> 00:18:49,040 HE THEN DID TRANSGENIC RESCUE 547 00:18:49,040 --> 00:18:49,560 BY 548 00:18:49,560 --> 00:18:55,800 PUTTING TIA INTO THE NEURONS 549 00:18:55,800 --> 00:19:01,800 WHICH FULLY RESCUE THE 550 00:19:01,800 --> 00:19:02,120 ASCENDED 551 00:19:02,120 --> 00:19:02,560 REGROWTH PHENOTYPE, 552 00:19:02,560 --> 00:19:03,360 BUT IT DOES NOT HAPPEN IF YOU 553 00:19:03,360 --> 00:19:03,560 ARE 554 00:19:03,560 --> 00:19:06,400 PUTTING THE GENES INTO MUSCLE 555 00:19:06,400 --> 00:19:07,000 CELLS AND ADDITIONALLY HE 556 00:19:07,000 --> 00:19:07,200 PLAYS 557 00:19:07,200 --> 00:19:11,200 THE HUMAN TIA INTO 558 00:19:11,200 --> 00:19:11,800 (INDISCERNIBLE) WHICH ALI 559 00:19:11,800 --> 00:19:12,000 FULLY 560 00:19:12,000 --> 00:19:16,840 RESCUED THE TIA NON-MUTANT 561 00:19:16,840 --> 00:19:17,160 PHENOTYPE. 562 00:19:17,160 --> 00:19:18,040 --- 563 00:19:18,040 --> 00:19:19,480 AND WHEN HE INCREASED THE 564 00:19:19,480 --> 00:19:22,920 EXPRESSION LEVEL OF TIAR-2, 565 00:19:22,920 --> 00:19:23,120 HE 566 00:19:23,120 --> 00:19:25,840 STARTED SEEING INHIBITION, A 567 00:19:25,840 --> 00:19:26,080 FACT 568 00:19:26,080 --> 00:19:27,400 THAT ALSO REDUCED AXON 569 00:19:27,400 --> 00:19:27,720 REGROWTH. 570 00:19:27,720 --> 00:19:36,600 SO THE TIAR PROTEIN BEHAVES 571 00:19:36,600 --> 00:19:36,840 AS A 572 00:19:36,840 --> 00:19:37,960 DOSAGE AND IS A FACTOR ON 573 00:19:37,960 --> 00:19:39,200 REGROWTH. 574 00:19:39,200 --> 00:19:41,600 AND IT IS ALSO EXHIBITING 575 00:19:41,600 --> 00:19:42,160 PATTERNS THAT ARE ALSO 576 00:19:42,160 --> 00:19:42,440 DEPENDENT 577 00:19:42,440 --> 00:19:44,360 ON DOSAGE. 578 00:19:44,360 --> 00:19:46,120 IN SOME CASES AND THE AXONS, 579 00:19:46,120 --> 00:19:47,000 THERE IS A SINGLE NEURON 580 00:19:47,000 --> 00:19:48,800 CALLED 581 00:19:48,800 --> 00:19:54,840 THE PLM, WITH A HIGH 582 00:19:54,840 --> 00:19:55,160 EXPRESSION 583 00:19:55,160 --> 00:19:56,600 LEVEL WILL FORM GRANULES AND 584 00:19:56,600 --> 00:19:56,800 AT 585 00:19:56,800 --> 00:19:57,560 LOW EXPRESSION IT IS DIFFUSED 586 00:19:57,560 --> 00:19:57,760 AND 587 00:19:57,760 --> 00:20:00,720 THIS CAN BE QUANTITATED HERE. 588 00:20:00,720 --> 00:20:02,520 THIS KIND OF A CONCENTRATION 589 00:20:02,520 --> 00:20:03,920 DEPENDS ON THE GRANULE 590 00:20:03,920 --> 00:20:04,240 FORMATION, 591 00:20:04,240 --> 00:20:05,320 RIGHT AROUND THE TIME CAME 592 00:20:05,320 --> 00:20:05,560 THIS 593 00:20:05,560 --> 00:20:13,080 IDEA OF LIQUIED PROTEIN 594 00:20:13,080 --> 00:20:13,360 PHASED 595 00:20:13,360 --> 00:20:18,000 OPERATIONS THAT COULD 596 00:20:18,000 --> 00:20:19,360 (INDISCERNIBLE) BEHAVIOR. 597 00:20:19,360 --> 00:20:26,760 AND MATT PURIFIED THE TIAR-2 598 00:20:26,760 --> 00:20:27,440 PROTEIN TO PERFORM THE LIQUID 599 00:20:27,440 --> 00:20:30,320 DROPLET ASSAY, WHICH HAS 600 00:20:30,320 --> 00:20:30,640 CROWDING 601 00:20:30,640 --> 00:20:31,720 FACTOR WHICH INSTANTLY SAW 602 00:20:31,720 --> 00:20:31,960 THERE 603 00:20:31,960 --> 00:20:34,640 IS INFORMATION OF THE 604 00:20:34,640 --> 00:20:34,920 DROPLETS, 605 00:20:34,920 --> 00:20:36,120 AND THOSE DROPLETS ONE OF THE 606 00:20:36,120 --> 00:20:37,160 BEHAVIORS IS THAT THEY HAVE 607 00:20:37,160 --> 00:20:37,840 LIQUID BEHAVIOR. 608 00:20:37,840 --> 00:20:43,560 WHEN HE FRACKED THEM AND THEY 609 00:20:43,560 --> 00:20:44,160 RECOVER VERY RAPIDLY. 610 00:20:44,160 --> 00:20:44,880 --- 611 00:20:44,880 --> 00:20:50,480 AND THIS TIAR-2 GRANULES, IN 612 00:20:50,480 --> 00:20:55,840 SUB-CULTURES THE TIAR PROTEIN 613 00:20:55,840 --> 00:20:56,040 IS 614 00:20:56,040 --> 00:20:58,560 BEEN CONSIDERED A CORE 615 00:20:58,560 --> 00:20:58,840 COMPONENT 616 00:20:58,840 --> 00:20:59,520 OF STRESS GRANULES INDUCED BY 617 00:20:59,520 --> 00:21:06,920 STRESS BUT IN C ELEGANS AXONS 618 00:21:06,920 --> 00:21:07,120 WE 619 00:21:07,120 --> 00:21:08,520 DID AN EXTENSIVE STUDY WHERE 620 00:21:08,520 --> 00:21:13,920 OTHER GRANULE MANUFACTURERS 621 00:21:13,920 --> 00:21:14,960 AND 622 00:21:14,960 --> 00:21:16,960 SYNOPTIC- PROTEINS, EXCEPT 623 00:21:16,960 --> 00:21:18,680 THE 624 00:21:18,680 --> 00:21:22,080 TIAR-2 ITSELF, GENERALLY 625 00:21:22,080 --> 00:21:22,400 THERE IS 626 00:21:22,400 --> 00:21:23,000 LITTLE COLOCALIZATION, 627 00:21:23,000 --> 00:21:26,960 INDICATING THAT THE TIAR-2 628 00:21:26,960 --> 00:21:28,160 PROTEIN IS RATHER 629 00:21:28,160 --> 00:21:28,520 HOMOGENEOUS, 630 00:21:28,520 --> 00:21:29,160 VERY DYNAMIC. 631 00:21:29,160 --> 00:21:32,040 --- 632 00:21:32,040 --> 00:21:32,800 AND WE SEE THIS DYNAMIC 633 00:21:32,800 --> 00:21:33,080 ASPECT IN 634 00:21:33,080 --> 00:21:33,560 THE SLIDE IMAGING. 635 00:21:33,560 --> 00:21:35,880 THAT IS WHERE YOU SEE THE 636 00:21:35,880 --> 00:21:37,200 TIAR-2 637 00:21:37,200 --> 00:21:38,640 GRANULES, BASICALLY COMING IN 638 00:21:38,640 --> 00:21:46,560 FUSED AND SPLIT, BUT THEY 639 00:21:46,560 --> 00:21:47,120 ACTUALLY NO LONG-RANGE 640 00:21:47,120 --> 00:21:47,560 TRANSPORT. 641 00:21:47,560 --> 00:21:48,840 --- 642 00:21:48,840 --> 00:21:49,480 AND ALSO RESPONDING TO INJURY 643 00:21:49,480 --> 00:21:53,800 THIS IS SPLIT AND MERGED AS 644 00:21:53,800 --> 00:21:54,920 SHOWN 645 00:21:54,920 --> 00:21:56,520 HERE AND PREINJURY THERE IS 646 00:21:56,520 --> 00:21:58,920 ONE 647 00:21:58,920 --> 00:22:00,200 CLUSTER AND THEN AFTER INJURY 648 00:22:00,200 --> 00:22:00,840 THIS TRIGGER SOMETIMES FEWER, 649 00:22:00,840 --> 00:22:01,480 SOMETIMES MORE. 650 00:22:01,480 --> 00:22:03,920 BUT THIS IS A VERY TREND IN 651 00:22:03,920 --> 00:22:07,040 EVENTS. MATT DECIDED TO 652 00:22:07,040 --> 00:22:07,280 ADDRESS 653 00:22:07,280 --> 00:22:09,360 THE MECHANISM OF WHAT DRIVES 654 00:22:09,360 --> 00:22:11,080 THIS 655 00:22:11,080 --> 00:22:13,000 TIAR-2 BEHAVIOR AND FOR THIS, 656 00:22:13,000 --> 00:22:16,360 THIS BASICALLY SUMMARIZED HIS 657 00:22:16,360 --> 00:22:20,400 TWO-YEARS' WORK, EXPRESSING 658 00:22:20,400 --> 00:22:21,880 GAP 659 00:22:21,880 --> 00:22:26,360 TALK, TIAR-2 INVIVO 660 00:22:26,360 --> 00:22:26,720 CONCENTRATION 661 00:22:26,720 --> 00:22:30,200 IN A NICELY CONTROLLED WAY, 662 00:22:30,200 --> 00:22:30,400 AND 663 00:22:30,400 --> 00:22:30,920 TAKING EACH DOMAINS IT 664 00:22:30,920 --> 00:22:31,280 PURIFIED 665 00:22:31,280 --> 00:22:34,440 THE PROTEINS AND 666 00:22:34,440 --> 00:22:34,840 (INDISCERNIBLE) 667 00:22:34,840 --> 00:22:37,320 DROPLET HAVEN'S AS WELL AS IN 668 00:22:37,320 --> 00:22:44,880 VIVO WHETHER IT FORMS 669 00:22:44,880 --> 00:22:45,160 GRANULES 670 00:22:45,160 --> 00:22:46,920 ARE NOT, 671 00:22:46,920 --> 00:22:47,920 AND IT CAN RESCUE AND 672 00:22:47,920 --> 00:22:50,000 COMPLEMENT 673 00:22:50,000 --> 00:22:51,920 AXON REGENERATION PHENOTYPE. 674 00:22:51,920 --> 00:22:53,520 AND THE SNAPSHOT IS THAT THE 675 00:22:53,520 --> 00:22:56,440 FULL 676 00:22:56,440 --> 00:22:59,640 LENS TIAR-2 FORMS GRANULES 677 00:22:59,640 --> 00:22:59,840 AND 678 00:22:59,840 --> 00:23:01,960 RESCUES THE ENHANCED AXON 679 00:23:01,960 --> 00:23:07,320 REGROWTH OF TIAR-2 GROWTH. 680 00:23:07,320 --> 00:23:12,480 AND WE DELETED ALL THE RM 681 00:23:12,480 --> 00:23:16,680 DOMAINS, AXONS STILL FORMED 682 00:23:16,680 --> 00:23:16,880 IN 683 00:23:16,880 --> 00:23:18,400 THE ACTIVITY OF RESCUING THE 684 00:23:18,400 --> 00:23:19,760 MUTANTS STILL THERE. 685 00:23:19,760 --> 00:23:21,560 BUT WHEN HE PERTURBED THE -- 686 00:23:21,560 --> 00:23:28,560 LIKE 687 00:23:28,560 --> 00:23:29,160 DOMAIN, EVEN A SMALL DEMAND 688 00:23:29,160 --> 00:23:30,000 WHICH 689 00:23:30,000 --> 00:23:33,800 ABOLISHED THE AXON GRANULE 690 00:23:33,800 --> 00:23:34,480 SO 691 00:23:34,480 --> 00:23:35,080 THERE IS A SMALL DOMAIN FOR 692 00:23:35,080 --> 00:23:35,280 THE 693 00:23:35,280 --> 00:23:37,600 NUCLEUS GRANULES PROVIDE THE 694 00:23:37,600 --> 00:23:43,440 TIAR-2 FUNCTION. 695 00:23:43,440 --> 00:23:44,120 AND I'M GOING INTO ADDITIONAL 696 00:23:44,120 --> 00:23:46,880 DETAILS. 697 00:23:46,880 --> 00:23:47,480 HE WAS ABLE TO DEFY BOTH A 698 00:23:47,480 --> 00:23:54,280 PHOSPHORYLATION EVENT IN THAT 699 00:23:54,280 --> 00:23:54,840 SMALL DOMAIN, RESPONSE TO 700 00:23:54,840 --> 00:23:55,120 TRIGGER 701 00:23:55,120 --> 00:23:55,720 OPERATIONS OR PICK THEM UP 702 00:23:55,720 --> 00:23:55,920 WITH 703 00:23:55,920 --> 00:24:06,480 THIS MODEL THAT IS IN AXONS, 704 00:24:10,840 --> 00:24:11,360 TIAR-2 PROTEINS ARE ALWAYS 705 00:24:11,360 --> 00:24:11,600 THERE 706 00:24:11,600 --> 00:24:12,160 AND THEY UNDERGO A VERY 707 00:24:12,160 --> 00:24:12,440 DYNAMIC 708 00:24:12,440 --> 00:24:13,240 CHANGE. 709 00:24:13,240 --> 00:24:13,840 (INDISCERNIBLE). 710 00:24:13,840 --> 00:24:15,920 --- 711 00:24:15,920 --> 00:24:19,960 THIS TEMPORAL GRANULE 712 00:24:19,960 --> 00:24:20,280 FORMATION 713 00:24:20,280 --> 00:24:23,120 IS A STAGE SAMPLING THE 714 00:24:23,120 --> 00:24:23,400 INJURY 715 00:24:23,400 --> 00:24:24,120 SIGNAL AND SPREADING THE 716 00:24:24,120 --> 00:24:25,480 SIGNAL 717 00:24:25,480 --> 00:24:26,200 INTO THE ENTIRE AXON. 718 00:24:26,200 --> 00:24:29,120 --- 719 00:24:29,120 --> 00:24:31,320 THIS STUDY WAS ABOUT THE SAME 720 00:24:31,320 --> 00:24:31,960 TIME, NICELY SYNERGIZE WITH 721 00:24:31,960 --> 00:24:32,280 JEFF 722 00:24:32,280 --> 00:24:37,960 -- WORK, USING MOUSE SCIATIC 723 00:24:37,960 --> 00:24:38,960 NERVE FOR THE IDENTIFIED A 724 00:24:38,960 --> 00:24:42,840 BINDING FACTOR OF TIA 725 00:24:42,840 --> 00:24:53,080 PROTEIN, 726 00:24:53,400 --> 00:24:55,040 FORMING GRANULES AND IN A 727 00:24:55,040 --> 00:24:55,640 PHOSPHORYLATION- DEPENDENT 728 00:24:55,640 --> 00:24:55,960 MANNER. 729 00:24:55,960 --> 00:24:56,920 UNDER THIS MECHANISM FROM C 730 00:24:56,920 --> 00:25:00,600 ELEGANS TO MAMMALS AND HUMANS 731 00:25:00,600 --> 00:25:00,800 AS 732 00:25:00,800 --> 00:25:01,400 WELL. 733 00:25:01,400 --> 00:25:07,160 --- 734 00:25:07,160 --> 00:25:07,720 SO THIS IS HOW WE WERE 735 00:25:07,720 --> 00:25:08,040 DISSECTING 736 00:25:08,040 --> 00:25:13,720 THE PROTEIN OF TIAR-2, 737 00:25:13,720 --> 00:25:14,240 AND THIS IS GOING TO MY 738 00:25:14,240 --> 00:25:14,560 FAVORITE 739 00:25:14,560 --> 00:25:18,280 MOLECULE, THE DLK-1, NAMED 740 00:25:18,280 --> 00:25:26,240 BECAUSE IT IS A MAPS 3 741 00:25:26,240 --> 00:25:27,720 KINASE, 742 00:25:27,720 --> 00:25:35,360 NOT ENTIRELY ACCURATE IN THE 743 00:25:35,360 --> 00:25:37,520 SENSE THAT IS IN 744 00:25:37,520 --> 00:25:39,000 (INDISCERNIBLE), 745 00:25:39,000 --> 00:25:42,920 CHARACTERIZED BY TWO SUPER 746 00:25:42,920 --> 00:25:46,680 DOMAINS ONE CALLED MAP3K12, 747 00:25:46,680 --> 00:25:49,800 ONE 748 00:25:49,800 --> 00:25:55,480 MAP3K13, THE 13 IS THE 749 00:25:55,480 --> 00:25:58,600 (INDISCERNIBLE) KINASE. 750 00:25:58,600 --> 00:26:01,960 THEY HAVE -- 751 00:26:01,960 --> 00:26:03,680 C ELEGANS HAS A SIMPLE NAME 752 00:26:03,680 --> 00:26:04,440 CALL 753 00:26:04,440 --> 00:26:06,720 THE DLK-1. 754 00:26:06,720 --> 00:26:07,160 AND APART FROM THIS 755 00:26:07,160 --> 00:26:07,880 CONSERVATION, 756 00:26:07,880 --> 00:26:10,920 ALL THESE KINASES ARE 757 00:26:10,920 --> 00:26:12,120 SUBJECTED 758 00:26:12,120 --> 00:26:14,960 TO (INDISCERNIBLE) REGULATED 759 00:26:14,960 --> 00:26:15,720 LIPID MODIFICATION. 760 00:26:15,720 --> 00:26:16,360 --- 761 00:26:16,360 --> 00:26:20,680 WE STUDIED THE DLK, LONG 762 00:26:20,680 --> 00:26:22,920 BEFORE 763 00:26:22,920 --> 00:26:25,640 WE ESTABLISHED LASER EXALTOMY 764 00:26:25,640 --> 00:26:28,160 ASSAY. 765 00:26:28,160 --> 00:26:30,680 AND WE ESTABLISH THIS, AS 766 00:26:30,680 --> 00:26:30,960 PART OF 767 00:26:30,960 --> 00:26:32,400 THE EARLY INTEREST IN MY LAB, 768 00:26:32,400 --> 00:26:32,600 HOW 769 00:26:32,600 --> 00:26:34,120 DO WE FORM SYNAPSES? 770 00:26:34,120 --> 00:26:42,440 WE USED THE -- LABELED GAP TO 771 00:26:42,440 --> 00:26:43,040 MARK THE SYNAPTIC TERMINAL, 772 00:26:43,040 --> 00:26:43,520 LOOKING FOR MUTANTS 773 00:26:43,520 --> 00:26:43,840 DISRUPTING 774 00:26:43,840 --> 00:26:48,400 THIS VERY NICE SYNOPTIC 775 00:26:48,400 --> 00:26:48,720 PUNCTURE, 776 00:26:48,720 --> 00:26:51,720 AND THIS LED TO A GENE CALLED 777 00:26:51,720 --> 00:26:54,920 RPM-1 REGULATOR PRESYNAPTIC 778 00:26:54,920 --> 00:26:56,000 MORPHOLOGY ONE, 779 00:26:56,000 --> 00:26:58,600 IN THIS CLUSTER BECOME VERY 780 00:26:58,600 --> 00:27:03,080 IRREGULAR COMPARED TO WILD 781 00:27:03,080 --> 00:27:04,280 TYPE 782 00:27:04,280 --> 00:27:05,920 NORMAL SHAPED 783 00:27:05,920 --> 00:27:06,360 (INDISCERNIBLE), 784 00:27:06,360 --> 00:27:07,320 AND THE ACTIVE ZONE PROTEIN 785 00:27:07,320 --> 00:27:11,680 CALLED LIPRIN. 786 00:27:11,680 --> 00:27:12,360 THIS ORGANIZED SYNOPTIC 787 00:27:12,360 --> 00:27:12,640 CLUSTER 788 00:27:12,640 --> 00:27:21,560 IN -- MUTANTS HAVE A LARGE 789 00:27:21,560 --> 00:27:21,840 SPREAD 790 00:27:21,840 --> 00:27:24,280 OF VESICOSE (INDISCERNIBLE). 791 00:27:24,280 --> 00:27:24,880 AND IT TURNS OUT THAT THIS 792 00:27:24,880 --> 00:27:27,520 PROTEIN -- FEELS LIKE MY 793 00:27:27,520 --> 00:27:29,080 VOICE IS 794 00:27:29,080 --> 00:27:32,880 NOT TRANSMITTING -- SO IT 795 00:27:32,880 --> 00:27:33,400 TURNS 796 00:27:33,400 --> 00:27:41,520 OUT THAT THE RPM-1 PROTEIN 797 00:27:41,520 --> 00:27:41,720 IS 798 00:27:41,720 --> 00:27:45,440 PART OF THE FAMILY CALLED PHR. 799 00:27:45,440 --> 00:27:47,800 AND IT BUILDS A PROTEIN 800 00:27:47,800 --> 00:27:49,120 CALLED 801 00:27:49,120 --> 00:27:49,440 "HIGH WIRE." 802 00:27:49,440 --> 00:27:50,720 THESE LARGE PROTEINS, CLOSE 803 00:27:50,720 --> 00:27:52,720 TO 804 00:27:52,720 --> 00:27:55,080 4000 IMMUNIZES (PHONETIC) 805 00:27:55,080 --> 00:27:55,360 MORE OR 806 00:27:55,360 --> 00:27:57,360 LESS 807 00:27:57,360 --> 00:27:57,960 THEY ALL COME FROM THE WORK 808 00:27:57,960 --> 00:27:58,160 OF 809 00:27:58,160 --> 00:27:59,320 CORY GOODMAN, (INDISCERNIBLE) 810 00:27:59,320 --> 00:27:59,800 -- 811 00:27:59,800 --> 00:28:05,400 ALL COME TOGETHER TO FORM 812 00:28:05,400 --> 00:28:10,440 GENETICS AND MOLECULARIZED 813 00:28:10,440 --> 00:28:11,000 APPROACHES SHOWING THAT 814 00:28:11,000 --> 00:28:13,400 EVERYONE 815 00:28:13,400 --> 00:28:14,000 PLACE IMPORTANT ROLES IN 816 00:28:14,000 --> 00:28:17,040 DIFFERENT STAGES OF AXON 817 00:28:17,040 --> 00:28:20,240 TERMINATION AND GUIDANCE, 818 00:28:20,240 --> 00:28:20,520 SETTING 819 00:28:20,520 --> 00:28:21,080 UP SYNAPSE STABILIZATION. 820 00:28:21,080 --> 00:28:21,720 --- 821 00:28:21,720 --> 00:28:25,840 A COMMON FEATURE OF THIS PHR 822 00:28:25,840 --> 00:28:27,160 PROTEIN IS THAT THIS SUGGESTS 823 00:28:27,160 --> 00:28:27,360 IT 824 00:28:27,360 --> 00:28:34,880 IS ACTUALLY AN EASERUBIC 825 00:28:34,880 --> 00:28:36,400 (PHONETIC) LIGASE. 826 00:28:36,400 --> 00:28:40,040 IT ACTS AS A SYNOPSIS. 827 00:28:40,040 --> 00:28:41,640 WHAT WOULD BE ITS SUBSTRATE 828 00:28:41,640 --> 00:28:41,840 AT 829 00:28:41,840 --> 00:28:43,560 THE SYNAPSES? 830 00:28:43,560 --> 00:28:45,280 FOR THIS MY LAB TOOK A 831 00:28:45,280 --> 00:28:45,560 GENETIC 832 00:28:45,560 --> 00:28:46,760 APPROACH, 833 00:28:46,760 --> 00:28:52,360 LOOKING FOR SUPPRESSORS OF 834 00:28:52,360 --> 00:28:52,720 RPM 835 00:28:52,720 --> 00:28:53,520 SYNAPTIC PHENOTYPES. 836 00:28:53,520 --> 00:28:58,480 IF THE (INDISCERNIBLE) IS NOT 837 00:28:58,480 --> 00:28:59,040 THERE SOME PROTEIN IS UP 838 00:28:59,040 --> 00:29:04,120 REGULATED, 839 00:29:04,120 --> 00:29:05,720 THEN YOU CAN KNOCK IT DOWN 840 00:29:05,720 --> 00:29:05,960 AND IT 841 00:29:05,960 --> 00:29:06,600 COULD RESTORE THE SIGNALING 842 00:29:06,600 --> 00:29:07,920 HOMEOSTASIS AND GETTING THE 843 00:29:07,920 --> 00:29:10,600 SUPPRESSOR MUTATION. 844 00:29:10,600 --> 00:29:13,040 THIS TURNED OUT TO BE A VERY 845 00:29:13,040 --> 00:29:13,560 EFFECTIVE SUPPRESSIVE 846 00:29:13,560 --> 00:29:13,840 MUTATION 847 00:29:13,840 --> 00:29:18,240 AND WE IDENTIFIED 848 00:29:18,240 --> 00:29:22,240 (INDISCERNIBLE) 849 00:29:22,240 --> 00:29:27,160 AND DLK-1 KINASE AS WELL AS 850 00:29:27,160 --> 00:29:28,560 THE 851 00:29:28,560 --> 00:29:29,200 DOWNSTREAM KINASE AND FINALLY 852 00:29:29,200 --> 00:29:29,360 A 853 00:29:29,360 --> 00:29:34,680 TRANSCRIPTION FACTOR OF THE 854 00:29:34,680 --> 00:29:34,880 LZ 855 00:29:34,880 --> 00:29:36,680 DOMAIN TRANSCRIPTION FACTOR. 856 00:29:36,680 --> 00:29:37,640 --- 857 00:29:37,640 --> 00:29:41,560 WE FURTHER SHOWED THAT THE 858 00:29:41,560 --> 00:29:41,760 DLK 859 00:29:41,760 --> 00:29:43,120 KINASE ITSELF IS A SUBSTRATE 860 00:29:43,120 --> 00:29:49,440 OF 861 00:29:49,440 --> 00:29:50,960 (INDISCERNIBLE) LIGASE. 862 00:29:50,960 --> 00:29:54,960 AND THIS IS VERY LOW IN 863 00:29:54,960 --> 00:29:55,320 ABUNDANCE. 864 00:29:55,320 --> 00:29:57,280 YOU SEE AN ABUNDANCE 865 00:29:57,280 --> 00:29:58,160 INCREASED IF 866 00:29:58,160 --> 00:29:58,520 YOU REMOVE IT. 867 00:29:58,520 --> 00:30:01,880 --- 868 00:30:01,880 --> 00:30:02,920 ALL OF THESE ARE BEAUTIFUL 869 00:30:02,920 --> 00:30:03,520 GENETICS AND WE ENJOYED IT 870 00:30:03,520 --> 00:30:03,720 VERY 871 00:30:03,720 --> 00:30:04,360 MUCH BUT THEN THERE IS THE 872 00:30:04,360 --> 00:30:06,240 QUESTION COMING IN. 873 00:30:06,240 --> 00:30:08,080 IT APPEARS THAT DURING 874 00:30:08,080 --> 00:30:09,760 DEVELOPMENT IN THE INITIAL 875 00:30:09,760 --> 00:30:13,160 FORMATION OF SYNOPSIS THAT 876 00:30:13,160 --> 00:30:15,600 DLK 877 00:30:15,600 --> 00:30:17,600 KINASE NEEDS TO BE RAPIDLY 878 00:30:17,600 --> 00:30:17,880 TURNED 879 00:30:17,880 --> 00:30:18,400 OVER TO MAKE SURE THE 880 00:30:18,400 --> 00:30:18,680 SYNAPSES 881 00:30:18,680 --> 00:30:19,280 FORM PROPERLY AND LOSS OF 882 00:30:19,280 --> 00:30:24,400 FUNCTION OF DLK IN ANY OF ITS 883 00:30:24,400 --> 00:30:29,680 DOWNSTREAM TARGETS IS NOT 884 00:30:29,680 --> 00:30:31,040 ESSENTIAL, AND THAT IS HOW WE 885 00:30:31,040 --> 00:30:31,240 CAN 886 00:30:31,240 --> 00:30:31,720 FIND THEM AS GENETIC 887 00:30:31,720 --> 00:30:32,080 SUPPRESSOR 888 00:30:32,080 --> 00:30:34,160 STRENGTH. THIS 889 00:30:34,160 --> 00:30:34,760 RAISES THE QUESTION WHAT IS 890 00:30:34,760 --> 00:30:34,960 THE 891 00:30:34,960 --> 00:30:35,520 IMPORTANT FUNCTION OF THE 892 00:30:35,520 --> 00:30:36,440 GENES? 893 00:30:36,440 --> 00:30:38,920 IT SEEMS THAT IT'S IMPORTANT 894 00:30:38,920 --> 00:30:39,560 IS 895 00:30:39,560 --> 00:30:41,920 DEGRADED ACTIVELY. 896 00:30:41,920 --> 00:30:42,560 UNTIL HE MADE THE CONNECTION 897 00:30:42,560 --> 00:30:42,920 WHEN 898 00:30:42,920 --> 00:30:45,160 WE WERE DOING THE LASER 899 00:30:45,160 --> 00:30:46,520 EXOLTOMY 900 00:30:46,520 --> 00:30:49,840 SCREEN AND REDISCOVER THE 901 00:30:49,840 --> 00:30:50,920 DLK, 902 00:30:50,920 --> 00:30:52,440 AND IT IS AWFULLY ESSENTIAL 903 00:30:52,440 --> 00:30:52,640 TO 904 00:30:52,640 --> 00:30:58,080 INITIATE THE INJURY INDUCED 905 00:30:58,080 --> 00:30:58,640 REGROWTH AS YOU SEE HERE 906 00:30:58,640 --> 00:30:59,600 COMPARED 907 00:30:59,600 --> 00:31:01,480 TO WELL-TYPE. (INDISCERNIBLE) 908 00:31:01,480 --> 00:31:01,840 AND 909 00:31:01,840 --> 00:31:12,080 LOSS OF DLK DON'T FORM, AND 910 00:31:12,080 --> 00:31:12,320 AXONS 911 00:31:12,320 --> 00:31:13,160 STOP GROWING. 912 00:31:13,160 --> 00:31:18,920 IF WE ELEVATE THE DLK, IT 913 00:31:18,920 --> 00:31:19,600 INCREASE THE REGROWTH 914 00:31:19,600 --> 00:31:19,960 COMPLETELY 915 00:31:19,960 --> 00:31:21,120 DEPENDENT ON ITS TARGET SUCH 916 00:31:21,120 --> 00:31:22,920 AS 917 00:31:22,920 --> 00:31:24,320 THIS TRANSCRIPTION FACTOR. 918 00:31:24,320 --> 00:31:26,040 BY ITSELF IT DID NOT 919 00:31:26,040 --> 00:31:26,400 REGENERATE, 920 00:31:26,400 --> 00:31:27,920 AND ALSO IT BLOCKS THE 921 00:31:27,920 --> 00:31:30,160 ENHANCED 922 00:31:30,160 --> 00:31:31,000 DLK OVEREXPRESSION EFFECT. 923 00:31:31,000 --> 00:31:32,880 --- 924 00:31:32,880 --> 00:31:34,040 BESIDES TRIGGERING THIS 925 00:31:34,040 --> 00:31:34,880 DEVELOPMENTAL SIGNALING 926 00:31:34,880 --> 00:31:42,200 CASCADE, 927 00:31:42,200 --> 00:31:42,720 DLK ALSO PROMOTE INJURY 928 00:31:42,720 --> 00:31:43,000 INDUCED 929 00:31:43,000 --> 00:31:46,760 MICROTUBULE DYNAMICS. YOU 930 00:31:46,760 --> 00:31:47,000 RECALL 931 00:31:47,000 --> 00:31:47,560 THAT WHEN WE USE BINDING 932 00:31:47,560 --> 00:31:47,840 FACTORS 933 00:31:47,840 --> 00:31:50,760 WE OBSERVED THERE IS INJURY 934 00:31:50,760 --> 00:31:54,760 INDUCED EBP, AND SUSTAINED 935 00:31:54,760 --> 00:31:55,240 EBP 936 00:31:55,240 --> 00:31:59,320 GROWTH AND -- BOTH ARE 937 00:31:59,320 --> 00:32:00,240 INDUCED 938 00:32:00,240 --> 00:32:06,120 FEWER -- GETS INDUCED. 939 00:32:06,120 --> 00:32:07,000 SUSTAINED GROWTH DOES NOT 940 00:32:07,000 --> 00:32:09,200 HAPPEN. 941 00:32:09,200 --> 00:32:09,840 COMBINED WITH OTHER GENES WE 942 00:32:09,840 --> 00:32:10,040 CAN 943 00:32:10,040 --> 00:32:11,080 SHOW PART OF THIS IS DUE TO 944 00:32:11,080 --> 00:32:13,880 DISRUPTIONS IN 945 00:32:13,880 --> 00:32:14,920 POST-TRANSLATION 946 00:32:14,920 --> 00:32:17,160 MODIFICATIONS SUCH AS 947 00:32:17,160 --> 00:32:17,920 TARASINATED 948 00:32:17,920 --> 00:32:18,920 TUBULINGS (PHONETIC). 949 00:32:18,920 --> 00:32:22,160 --- 950 00:32:22,160 --> 00:32:22,920 THERE IS A LOT OF INTEREST AT 951 00:32:22,920 --> 00:32:26,880 THIS STAGE LINKING THIS DLK, 952 00:32:26,880 --> 00:32:28,600 WORKING ALSO ON DLK-1 AND -- 953 00:32:28,600 --> 00:32:29,000 -- 954 00:32:29,000 --> 00:32:39,440 WORK ON NUTRACELL DLK. 955 00:32:42,720 --> 00:32:47,080 THE - ACTION HAS TWO 956 00:32:47,080 --> 00:32:49,600 HOMOLOGUES, 957 00:32:49,600 --> 00:32:50,120 BOTH EXPRESSING IN THE 958 00:32:50,120 --> 00:32:50,400 NERVOUS 959 00:32:50,400 --> 00:32:54,320 SYSTEM AND CLONED FROM 960 00:32:54,320 --> 00:32:56,080 (INDISCERNIBLE) -- BOTH 961 00:32:56,080 --> 00:32:57,160 KINASE 962 00:32:57,160 --> 00:33:03,160 CAN ACT TO -- SIGNALING. 963 00:33:03,160 --> 00:33:10,680 THERE ARE MULTIPLE DLK MICE, 964 00:33:10,680 --> 00:33:18,440 GENERALLY DIE PERINATALLY. 965 00:33:18,440 --> 00:33:20,920 IF THEY ARE DELETED IN 966 00:33:20,920 --> 00:33:21,200 ADULTS, 967 00:33:21,200 --> 00:33:21,720 AND THIS IS WHERE YOUR 968 00:33:21,720 --> 00:33:22,000 SCIENCE 969 00:33:22,000 --> 00:33:31,800 DEPARTMENTS -- THOSE DLK MICE 970 00:33:31,800 --> 00:33:34,680 ARE 971 00:33:34,680 --> 00:33:35,320 PERFECTLY FUNCTIONING ADULTS. 972 00:33:35,320 --> 00:33:37,000 --- 973 00:33:37,000 --> 00:33:39,720 THERE IS A STUDY -- PERFECTLY 974 00:33:39,720 --> 00:33:45,000 VIABLE, NO DETECTABLE DEFECTS. 975 00:33:45,000 --> 00:33:48,280 BUT IN THE LAST 10 YEARS, 976 00:33:48,280 --> 00:33:48,480 INDIA 977 00:33:48,480 --> 00:33:54,600 FROM USING MAMMALIAN 978 00:33:54,600 --> 00:33:55,240 AXON-REGENERATION RESEARCHERS 979 00:33:55,240 --> 00:33:55,880 PARTICULARLY FROM OPTIC NERVE 980 00:33:55,880 --> 00:33:56,160 INJURIES, 981 00:33:56,160 --> 00:33:58,120 FROM A GENETIC GROUP AS WELL 982 00:33:58,120 --> 00:33:59,480 AS 983 00:33:59,480 --> 00:34:02,960 THE JOHNS HOPKINS GROUP, 984 00:34:02,960 --> 00:34:03,520 AND SOME PEOPLE USING THE 985 00:34:03,520 --> 00:34:04,560 PERIPHERY NERVE INJURY -- 986 00:34:04,560 --> 00:34:05,120 HAVE 987 00:34:05,120 --> 00:34:11,240 ALL SHOWN THAT THE DLK KINASE 988 00:34:11,240 --> 00:34:11,440 IS 989 00:34:11,440 --> 00:34:12,400 IMPORTANT FOR OPTIC NERVE 990 00:34:12,400 --> 00:34:13,000 INJURY 991 00:34:13,000 --> 00:34:20,520 INDUCED AXON SPROUTING; OPTIC 992 00:34:20,520 --> 00:34:26,280 NERVE -- SURVIVAL AS WELL AS 993 00:34:26,280 --> 00:34:26,920 REGENERATION EFFECT IN MY LAB 994 00:34:26,920 --> 00:34:27,120 IN 995 00:34:27,120 --> 00:34:29,520 COLLABORATION WITH -- GROUP 996 00:34:29,520 --> 00:34:30,960 AT 997 00:34:30,960 --> 00:34:34,960 UCSD DECIDED TO FOCUS ON DLK 998 00:34:34,960 --> 00:34:37,160 KINASE SHOWING THAT IT HAS 999 00:34:37,160 --> 00:34:44,680 STRONGER ROLE IN ASTRO CIZ3 1000 00:34:44,680 --> 00:34:50,440 (PHONETIC) -- INDUCED BY 1001 00:34:50,440 --> 00:34:51,080 INJURY 1002 00:34:51,080 --> 00:34:57,520 INVOLVING -- AND AT NIH HAVE 1003 00:34:57,520 --> 00:34:57,760 ALSO 1004 00:34:57,760 --> 00:35:04,080 IMPLICATED DLK'S ROLE IN HER 1005 00:35:04,080 --> 00:35:04,640 CAREER AND HER EARLY WORK 1006 00:35:04,640 --> 00:35:08,400 (INDISCERNIBLE) THE DLK 1007 00:35:08,400 --> 00:35:10,000 KINASE IS 1008 00:35:10,000 --> 00:35:11,560 PROTECTIVE IN ANIMAL MODELS 1009 00:35:11,560 --> 00:35:12,760 FOR 1010 00:35:12,760 --> 00:35:13,360 NEURODEGENERATIVE DISEASES 1011 00:35:13,360 --> 00:35:13,680 SUCH 1012 00:35:13,680 --> 00:35:14,680 AS ALZHEIMER'S AND ALS. 1013 00:35:14,680 --> 00:35:16,160 --- 1014 00:35:16,160 --> 00:35:18,560 IN RECENT YEARS, THROUGH 1015 00:35:18,560 --> 00:35:21,880 COLLABORATION WITH -- GROUP, 1016 00:35:21,880 --> 00:35:22,080 WE 1017 00:35:22,080 --> 00:35:23,640 ADDRESS THE FACT THAT DOES 1018 00:35:23,640 --> 00:35:24,000 THE 1019 00:35:24,000 --> 00:35:31,080 DLK AND LZK HAVE ANY ROLE IN 1020 00:35:31,080 --> 00:35:34,360 SPINAL CORD INJURY? 1021 00:35:34,360 --> 00:35:35,560 THERE ARE EXTENSIVE MOUSE 1022 00:35:35,560 --> 00:35:36,720 GENETICS; WE WILL HAVE TO USE 1023 00:35:36,720 --> 00:35:37,040 THE 1024 00:35:37,040 --> 00:35:41,200 P10 AS A BACKGROUND. 1025 00:35:41,200 --> 00:35:43,880 IN THE SPINAL CORD WHICH IS A 1026 00:35:43,880 --> 00:35:47,720 CENTRAL NERVOUS SYSTEM, AXONS 1027 00:35:47,720 --> 00:35:52,960 DON'T REGROW UNLESS YOU ARE 1028 00:35:52,960 --> 00:35:54,960 -- 1029 00:35:54,960 --> 00:35:56,480 AND STARTS SPROUTING, THAT 1030 00:35:56,480 --> 00:35:56,680 WAS 1031 00:35:56,680 --> 00:35:57,880 QUITE A LOT OF WORK. 1032 00:35:57,880 --> 00:36:03,000 IF THIS FIGURE IS TOO MUCH TO 1033 00:36:03,000 --> 00:36:04,520 LOOK, THE IMPORTANT THING IS 1034 00:36:04,520 --> 00:36:04,760 THAT 1035 00:36:04,760 --> 00:36:05,280 THIS IS THE CONTROL, 1036 00:36:05,280 --> 00:36:05,600 ESSENTIALLY 1037 00:36:05,600 --> 00:36:10,560 FOLLOWING DORSAL HEMI 1038 00:36:10,560 --> 00:36:10,920 TRANSECTION 1039 00:36:10,920 --> 00:36:11,960 (PHONETIC), 1040 00:36:11,960 --> 00:36:18,080 AND NO SPROUTING -- -- BUT 1041 00:36:18,080 --> 00:36:24,160 REMOVING DLK AND LZK 1042 00:36:24,160 --> 00:36:24,480 ESSENTIALLY 1043 00:36:24,480 --> 00:36:25,800 THERE IS LITTLE SPROUTING TO 1044 00:36:25,800 --> 00:36:29,320 BLOCK THE P10 EFFECT. 1045 00:36:29,320 --> 00:36:34,560 INDIVIDUALLY DLK AND LZK DID 1046 00:36:34,560 --> 00:36:34,760 NOT 1047 00:36:34,760 --> 00:36:37,080 HAVE THIS STRONG EFFECT. 1048 00:36:37,080 --> 00:36:39,440 SO THEY ARE BLOCKING SPINAL 1049 00:36:39,440 --> 00:36:40,920 CORD 1050 00:36:40,920 --> 00:36:43,320 AXON REGENERATION AND INDUCED 1051 00:36:43,320 --> 00:36:44,160 (INDISCERNIBLE) DELETION. 1052 00:36:44,160 --> 00:36:46,880 --- 1053 00:36:46,880 --> 00:36:47,480 THIS IS ALL GOOD TO SUPPORT 1054 00:36:47,480 --> 00:36:47,720 THAT 1055 00:36:47,720 --> 00:36:54,480 THE WHOLE -- OF THE DLK 1056 00:36:54,480 --> 00:36:54,760 KINASE 1057 00:36:54,760 --> 00:36:55,840 ALONG WITH HIS SISTER KINASE 1058 00:36:55,840 --> 00:36:56,040 ARE 1059 00:36:56,040 --> 00:36:56,680 IMPORTANT FOR INJURY RESPONSE 1060 00:36:56,680 --> 00:37:01,040 FROM C ELEGANS TO MAMMALS. 1061 00:37:01,040 --> 00:37:02,760 AT THE SAME TIME YOU HAVE TO 1062 00:37:02,760 --> 00:37:04,720 THINK ABOUT, 1063 00:37:04,720 --> 00:37:05,560 IT IS NOT JUST TO BLOCK 1064 00:37:05,560 --> 00:37:05,840 THINGS, 1065 00:37:05,840 --> 00:37:07,040 YOU HAVE TO THINK ABOUT 1066 00:37:07,040 --> 00:37:07,720 ACTIVE 1067 00:37:07,720 --> 00:37:10,920 DLK SO YOU CAN PROMOTE AXONS 1068 00:37:10,920 --> 00:37:12,320 TO 1069 00:37:12,320 --> 00:37:13,880 REPAIR, REGROW AND DO 1070 00:37:13,880 --> 00:37:16,360 SOMETHING 1071 00:37:16,360 --> 00:37:18,240 BETTER. IN THIS CASE IT WAS 1072 00:37:18,240 --> 00:37:18,920 INTERESTING TO MENTION THESE 1073 00:37:18,920 --> 00:37:19,120 TWO 1074 00:37:19,120 --> 00:37:20,200 STUDIES. 1075 00:37:20,200 --> 00:37:22,640 ONE THIS WORK FROM THE UCLA 1076 00:37:22,640 --> 00:37:22,880 GROUP 1077 00:37:22,880 --> 00:37:26,000 SHOWING THAT IN THE 1078 00:37:26,000 --> 00:37:26,360 PARTICULAR 1079 00:37:26,360 --> 00:37:29,320 CONDITION, THE STROKE MODEL, 1080 00:37:29,320 --> 00:37:32,840 INHIBITING CCR5, CYTOKINE 1081 00:37:32,840 --> 00:37:36,480 RECEPTOR, IT HELPED THE MOUSE 1082 00:37:36,480 --> 00:37:36,680 TO 1083 00:37:36,680 --> 00:37:39,080 RECOVER FROM THE STROKE DUE 1084 00:37:39,080 --> 00:37:39,280 TO 1085 00:37:39,280 --> 00:37:40,080 THE FACT THAT THEY WERE ABLE 1086 00:37:40,080 --> 00:37:40,280 TO 1087 00:37:40,280 --> 00:37:43,880 SHOW THAT BLOCKING CCR5 1088 00:37:43,880 --> 00:37:44,160 FOLLOWING 1089 00:37:44,160 --> 00:37:47,000 THE STROKE AND DLK IS 1090 00:37:47,000 --> 00:37:54,080 UPREGULATED, TRIGGERING CROSS 1091 00:37:54,080 --> 00:37:56,760 SIGNALING WHICH TRIGGERS 1092 00:37:56,760 --> 00:37:58,520 (INDISCERNIBLE), AND SOME 1093 00:37:58,520 --> 00:38:00,840 PATIENTS WHO HAVE RECEIVED 1094 00:38:00,840 --> 00:38:01,080 CCR5 1095 00:38:01,080 --> 00:38:03,760 INHIBITORS ALSO RECOVER WELL. 1096 00:38:03,760 --> 00:38:08,240 IN THIS CCR5 STROKE MODEL 1097 00:38:08,240 --> 00:38:09,600 ACTIVE 1098 00:38:09,600 --> 00:38:18,200 DLK HELD THE MOTOR AXON 1099 00:38:18,200 --> 00:38:19,200 PLASTICITY TO HELP RECOVER 1100 00:38:19,200 --> 00:38:19,440 FROM 1101 00:38:19,440 --> 00:38:20,960 STROKE. 1102 00:38:20,960 --> 00:38:23,200 AND THIS ADDRESSES USING 1103 00:38:23,200 --> 00:38:23,440 HUMAN 1104 00:38:23,440 --> 00:38:27,160 INDUCED STEM CELLS, 1105 00:38:27,160 --> 00:38:27,480 ADDRESSING 1106 00:38:27,480 --> 00:38:30,640 HOW HUMAN INDUCED, HUMAN CELL 1107 00:38:30,640 --> 00:38:31,960 INDUCED NEURONS ARE 1108 00:38:31,960 --> 00:38:36,120 RESPONDING TO 1109 00:38:36,120 --> 00:38:37,920 APOE FACTORS WHICH IS A MAJOR 1110 00:38:37,920 --> 00:38:38,520 TRIGGER FOR ALZHEIMER'S AND 1111 00:38:38,520 --> 00:38:38,720 BY 1112 00:38:38,720 --> 00:38:46,840 APPLYING THE APOE4, THE CELLS 1113 00:38:46,840 --> 00:38:47,680 RESPOND POTENTLY TO 1114 00:38:47,680 --> 00:38:49,000 UPREGULATED 1115 00:38:49,000 --> 00:38:53,200 DLK KINASE AND THERE IS A 1116 00:38:53,200 --> 00:38:54,400 TRANSCRIPTION UP REGULATION 1117 00:38:54,400 --> 00:38:54,600 OF 1118 00:38:54,600 --> 00:38:56,320 APP. 1119 00:38:56,320 --> 00:39:02,360 IN OTHER WORDS -- IS UP 1120 00:39:02,360 --> 00:39:05,280 REGULATING DLK, AND 1121 00:39:05,280 --> 00:39:06,000 DEPENDING ON 1122 00:39:06,000 --> 00:39:06,600 WHAT SYSTEM YOU ARE SETTING 1123 00:39:06,600 --> 00:39:06,840 UP IT 1124 00:39:06,840 --> 00:39:07,600 COULD BE GOOD PROMOTING AXON 1125 00:39:07,600 --> 00:39:11,400 SPROUTING -- AND PROMOTING 1126 00:39:11,400 --> 00:39:11,600 APP 1127 00:39:11,600 --> 00:39:13,280 PRODUCTION WHICH IS 1128 00:39:13,280 --> 00:39:13,640 CONSISTENT 1129 00:39:13,640 --> 00:39:15,680 WITH CLARIS'S WORK, 1130 00:39:15,680 --> 00:39:16,840 INHIBITING 1131 00:39:16,840 --> 00:39:24,360 DLK CAN REDUCE NEURO- 1132 00:39:24,360 --> 00:39:34,840 REGENERATIVE PHENOTYPES. 1133 00:39:44,200 --> 00:39:50,040 --- WE ARE EXAMINING MICE AND 1134 00:39:50,040 --> 00:39:50,640 (INDISCERNIBLE) EXPRESSION 1135 00:39:50,640 --> 00:39:51,640 FOCUSING ON THE CEREBELLUM, 1136 00:39:51,640 --> 00:39:54,640 AND WE HAVE OUR COLLEAGUES TO 1137 00:39:54,640 --> 00:39:59,080 HELP US LEARN THE MOUSE 1138 00:39:59,080 --> 00:39:59,360 ANATOMY. 1139 00:39:59,360 --> 00:40:01,520 SUFFICE IT TO SAY THAT AFTER 1140 00:40:01,520 --> 00:40:01,720 THE 1141 00:40:01,720 --> 00:40:04,720 CONDITIONAL OUTCOME OF DLK 1142 00:40:04,720 --> 00:40:07,440 AND 1143 00:40:07,440 --> 00:40:10,280 LZK, BASICALLY THE CEREBELLUM 1144 00:40:10,280 --> 00:40:15,080 DEVELOPED QUITE NORMAL. 1145 00:40:15,080 --> 00:40:17,680 NO DEFECTS WE CAN DETECT. 1146 00:40:17,680 --> 00:40:20,840 YUNBO AND ERIN REGENERATED 1147 00:40:20,840 --> 00:40:22,520 THE 1148 00:40:22,520 --> 00:40:24,480 TRANSGENIC DLK, ALLOWING US 1149 00:40:24,480 --> 00:40:24,680 TO 1150 00:40:24,680 --> 00:40:27,760 MANIPULATE THE CELL TYPES AND 1151 00:40:27,760 --> 00:40:27,960 WE 1152 00:40:27,960 --> 00:40:33,520 WOULD CROSS THEM WITH -- IN 1153 00:40:33,520 --> 00:40:34,200 THIS 1154 00:40:34,200 --> 00:40:39,920 CASE DLK IS UPREGULATED TWO- 1155 00:40:39,920 --> 00:40:42,480 TO 1156 00:40:42,480 --> 00:40:46,960 FOUR-FOLD, UP REGULATING 1157 00:40:46,960 --> 00:40:47,240 SIGNALING, 1158 00:40:47,240 --> 00:40:47,960 AND MOST OF THE NEURONS 1159 00:40:47,960 --> 00:40:48,240 BASICALLY 1160 00:40:48,240 --> 00:40:55,440 DIE AND BY P21 MICE ALSO DIE 1161 00:40:55,440 --> 00:40:56,040 PERHAPS DUE TO ITS EFFECT IN 1162 00:40:56,040 --> 00:40:56,480 OTHER NEURONS. 1163 00:40:56,480 --> 00:40:57,800 --- 1164 00:40:57,800 --> 00:41:01,480 IN CONTRAST, WE MADE THE SAME 1165 00:41:01,480 --> 00:41:06,480 KIND OF MICE, THIS TIME 1166 00:41:06,480 --> 00:41:09,880 INDUCIBLE 1167 00:41:09,880 --> 00:41:16,680 OVEREXPRESSING LZK, AND THOSE 1168 00:41:16,680 --> 00:41:17,320 MICE CAN LIVE HAPPILY AFTER A 1169 00:41:17,320 --> 00:41:18,760 YEAR -- (INDISCERNIBLE) -- 1170 00:41:18,760 --> 00:41:19,360 LIVE 1171 00:41:19,360 --> 00:41:24,520 UP TO FOUR MONTHS, 40-60% 1172 00:41:24,520 --> 00:41:26,280 DEATHS. 1173 00:41:26,280 --> 00:41:31,160 AND LZK OVEREXPRESSION ALSO 1174 00:41:31,160 --> 00:41:33,720 INDUCED JUNK SIGNALING AND 1175 00:41:33,720 --> 00:41:33,960 THAT 1176 00:41:33,960 --> 00:41:37,600 IS INDUCED IN THE CUSP SPACE. 1177 00:41:37,600 --> 00:41:38,800 WE STARTED ASKING THE 1178 00:41:38,800 --> 00:41:39,080 QUESTION, 1179 00:41:39,080 --> 00:41:46,600 HOW CAN THESE TWO KINASES 1180 00:41:46,600 --> 00:41:48,280 SEEM TO 1181 00:41:48,280 --> 00:41:50,560 BEHAVE THE SAME? 1182 00:41:50,560 --> 00:41:54,000 WHY DID THEY APPEAR TO HAVE 1183 00:41:54,000 --> 00:41:54,960 DIFFERENT POTENCIES IN NEURON 1184 00:41:54,960 --> 00:41:56,200 SURVIVAL? 1185 00:41:56,200 --> 00:41:58,440 A LOT OF THIS IS GENERIC 1186 00:41:58,440 --> 00:41:58,680 THING. 1187 00:41:58,680 --> 00:42:01,520 TRIPLE KINASE WILL ACTIVATE 1188 00:42:01,520 --> 00:42:01,880 THE 1189 00:42:01,880 --> 00:42:06,240 DOUBLE KINASE AND GO THROUGH 1190 00:42:06,240 --> 00:42:06,440 THE 1191 00:42:06,440 --> 00:42:07,040 JUNK SIGNALING TO STRESS 1192 00:42:07,040 --> 00:42:08,120 KINASE. 1193 00:42:08,120 --> 00:42:10,680 IN THINKING ABOUT THE TWO 1194 00:42:10,680 --> 00:42:11,000 KINASE 1195 00:42:11,000 --> 00:42:14,360 DIFFERENCES, WE READ THIS 1196 00:42:14,360 --> 00:42:14,960 PAPER 1197 00:42:14,960 --> 00:42:18,920 WHICH ACTUALLY STUDIED T CELL 1198 00:42:18,920 --> 00:42:19,960 ACTIVATION. IN THIS PROCESS 1199 00:42:19,960 --> 00:42:20,200 JUNK 1200 00:42:20,200 --> 00:42:23,120 SIGNALING IS WIDESPREAD, AND 1201 00:42:23,120 --> 00:42:23,320 ONE 1202 00:42:23,320 --> 00:42:24,640 OF THE JUNK SIGNALING IS 1203 00:42:24,640 --> 00:42:25,880 INDUCED 1204 00:42:25,880 --> 00:42:28,880 BY MANY GENES INVOLVED IN T 1205 00:42:28,880 --> 00:42:29,200 CELL 1206 00:42:29,200 --> 00:42:31,480 DIFFERENTIATION. 1207 00:42:31,480 --> 00:42:32,680 THIS GROUP IDENTIFIED THE 1208 00:42:32,680 --> 00:42:34,960 MECHANISM TO SHOW THAT 1209 00:42:34,960 --> 00:42:45,440 THE JUNK UPSTREAM KINASE, 1210 00:42:49,920 --> 00:42:54,080 MKK7, DURING CHELSEA 1211 00:42:54,080 --> 00:42:56,000 (CORRECTION) 1212 00:42:56,000 --> 00:42:57,560 DURING T-CELL ACTIVATION. 1213 00:42:57,560 --> 00:42:58,560 (INDISCERNIBLE). 1214 00:42:58,560 --> 00:43:08,120 THE PRESENCE OF CELF2 1215 00:43:08,120 --> 00:43:08,440 SPLICING 1216 00:43:08,440 --> 00:43:09,000 FACTOR WILL INCLUDE THIS 1217 00:43:09,000 --> 00:43:11,440 ALTERNATIVE AXON 1218 00:43:11,440 --> 00:43:12,840 WILL HAVE THIS DOUBLE KINASE 1219 00:43:12,840 --> 00:43:16,280 AND IT WILL RESULT IN A LESS 1220 00:43:16,280 --> 00:43:17,680 ACTIVE MKK7. 1221 00:43:17,680 --> 00:43:19,120 --- 1222 00:43:19,120 --> 00:43:21,000 SO WHAT WE HAPPENED TO 1223 00:43:21,000 --> 00:43:21,320 REALIZE IS 1224 00:43:21,320 --> 00:43:28,400 THAT THERE IS A CELF 1225 00:43:28,400 --> 00:43:30,560 HOMOLOGOUS 1226 00:43:30,560 --> 00:43:31,240 2, IN BINDING FACTORS WITH 1227 00:43:31,240 --> 00:43:32,280 THE 3 1228 00:43:32,280 --> 00:43:38,400 RM DOMAINS, MOSTLY NUCLEAR, 1229 00:43:38,400 --> 00:43:39,280 NAMED 1230 00:43:39,280 --> 00:43:40,400 (INDISCERNIBLE) BINDING 1231 00:43:40,400 --> 00:43:41,640 FACTOR. 1232 00:43:41,640 --> 00:43:46,040 AND THE WORK OF MY FORMER 1233 00:43:46,040 --> 00:43:47,840 POST 1234 00:43:47,840 --> 00:43:54,720 DOC LIZHEN CHEN WHO IS NOW 1235 00:43:54,720 --> 00:44:01,120 PROFESSOR AT -- SHE OBSERVED 1236 00:44:01,120 --> 00:44:05,640 LOSING UNC-75, AXONS DON'T 1237 00:44:05,640 --> 00:44:05,880 GROW 1238 00:44:05,880 --> 00:44:07,200 MUCH, 1239 00:44:07,200 --> 00:44:11,440 NOT AS SEVERELY IMPAIRED AS A 1240 00:44:11,440 --> 00:44:11,640 DLK 1241 00:44:11,640 --> 00:44:12,280 LOSS OF FUNCTION BUT IT WILL 1242 00:44:12,280 --> 00:44:13,240 REDUCE THE REGROWTH. 1243 00:44:13,240 --> 00:44:17,240 SHE ALSO DECIDED TO VENTURE 1244 00:44:17,240 --> 00:44:17,800 INTO 1245 00:44:17,800 --> 00:44:28,320 -- FIELD AND GOT THE CELL TO 1246 00:44:32,320 --> 00:44:32,840 (INDISCERNIBLE) AND LOOKING 1247 00:44:32,840 --> 00:44:33,040 AT 1248 00:44:33,040 --> 00:44:33,520 THE SCIATIC NERVE 1249 00:44:33,520 --> 00:44:34,920 REGENERATION. 1250 00:44:34,920 --> 00:44:37,560 SHE BASICALLY DELETED THE 1251 00:44:37,560 --> 00:44:38,640 CELF2 1252 00:44:38,640 --> 00:44:44,680 FROM DRG AXONS MARKED IN RED. 1253 00:44:44,680 --> 00:44:48,720 THESE REGENERATING AXONS, 1254 00:44:48,720 --> 00:44:49,920 FOLLOWING THE CRASH AND THE 1255 00:44:49,920 --> 00:44:55,720 SCIATIC NERVE IS MARKED BY A 1256 00:44:55,720 --> 00:44:59,360 CG 1257 00:44:59,360 --> 00:45:01,680 10, AND FOLLOWING THIS CRASH, 1258 00:45:01,680 --> 00:45:05,000 THIS WOULD REGENERATE QUITE 1259 00:45:05,000 --> 00:45:05,600 WELL, 1260 00:45:05,600 --> 00:45:07,520 LOSING (INDISCERNIBLE) AND 1261 00:45:07,520 --> 00:45:17,400 REDUCING RATE GROWTH. 1262 00:45:17,400 --> 00:45:18,040 LOOKING FOR SPLICING TARGETS 1263 00:45:18,040 --> 00:45:20,840 BEGINNING TO IDENTIFY A 1264 00:45:20,840 --> 00:45:22,920 SERIES OF 1265 00:45:22,920 --> 00:45:23,880 TARGETS WHICH INCLUDED 1266 00:45:23,880 --> 00:45:24,200 SYNAPTIC 1267 00:45:24,200 --> 00:45:25,080 TRANSMISSION GENES ACROSS. 1268 00:45:25,080 --> 00:45:27,320 --- 1269 00:45:27,320 --> 00:45:28,080 NOW WE CAN START PUTTING 1270 00:45:28,080 --> 00:45:28,360 TOGETHER 1271 00:45:28,360 --> 00:45:33,000 THE C ELEGANS AND THE MOUSE 1272 00:45:33,000 --> 00:45:33,640 WORK 1273 00:45:33,640 --> 00:45:36,040 TO ASK THE QUESTION COULD 1274 00:45:36,040 --> 00:45:36,280 CELF2 1275 00:45:36,280 --> 00:45:41,320 INFLUENCE THE DLK AND LZK 1276 00:45:41,320 --> 00:45:41,560 EFFECT 1277 00:45:41,560 --> 00:45:42,760 THROUGH ALTERNATIVE SPLICING 1278 00:45:42,760 --> 00:45:42,960 OF 1279 00:45:42,960 --> 00:45:46,520 MKK7? 1280 00:45:46,520 --> 00:45:47,960 FOR THIS WE MADE EXTENSIVE 1281 00:45:47,960 --> 00:45:53,440 GENETIC MOUSE -- COMBINATIONS. 1282 00:45:53,440 --> 00:45:55,880 AND ADDITIONAL DELETING CELL 1283 00:45:55,880 --> 00:45:56,080 IN 1284 00:45:56,080 --> 00:46:00,280 THE DLK OVEREXPRESSION MICE, 1285 00:46:00,280 --> 00:46:00,480 AND 1286 00:46:00,480 --> 00:46:03,520 ASKING TODAY BLOCK THE 1287 00:46:03,520 --> 00:46:05,000 CONTINUING 1288 00:46:05,000 --> 00:46:07,840 CELL DEATHS. 1289 00:46:07,840 --> 00:46:08,720 THERE IS NO EFFECT WHATSOEVER. 1290 00:46:08,720 --> 00:46:13,440 BUT WITH LZK OVEREXPRESSION 1291 00:46:13,440 --> 00:46:13,640 WE 1292 00:46:13,640 --> 00:46:14,160 SEE A VERY SIGNIFICANT 1293 00:46:14,160 --> 00:46:19,520 SUPPRESSION OF LZK-INDUCED 1294 00:46:19,520 --> 00:46:25,240 PURKINJE CELL DEATHS. 1295 00:46:25,240 --> 00:46:25,800 THOSE ANIMALS LIVE MUCH 1296 00:46:25,800 --> 00:46:26,040 LONGER 1297 00:46:26,040 --> 00:46:26,400 AND HAPPIER. 1298 00:46:26,400 --> 00:46:27,400 --- 1299 00:46:27,400 --> 00:46:27,960 THROUGH EXAMINING SIGNALS 1300 00:46:27,960 --> 00:46:30,040 FROM C 1301 00:46:30,040 --> 00:46:33,640 ELEGANS TO MAMMALS, WE ARE 1302 00:46:33,640 --> 00:46:34,280 FILLING IN ONE PART OF THE 1303 00:46:34,280 --> 00:46:37,680 KNOWLEDGE THAT DLK PROBABLY 1304 00:46:37,680 --> 00:46:43,080 ACTING THROUGH A DIFFERENT -- 1305 00:46:43,080 --> 00:46:47,120 CALLED MKK4 AND 1306 00:46:47,120 --> 00:46:48,400 (INDISCERNIBLE) 1307 00:46:48,400 --> 00:46:57,800 WHERE LZK ACCESS THROUGH MKK7 1308 00:46:57,800 --> 00:46:58,000 AND 1309 00:46:58,000 --> 00:46:58,520 TRIGGERS A SLOW JUNK 1310 00:46:58,520 --> 00:46:58,800 SIGNALING 1311 00:46:58,800 --> 00:46:59,400 LEADING TO (INDISCERNIBLE) 1312 00:46:59,400 --> 00:47:00,360 AND 1313 00:47:00,360 --> 00:47:03,080 BOTH KINASES EVEN THOUGH THEY 1314 00:47:03,080 --> 00:47:07,880 BEHAVE THE SAME IN NEURONS 1315 00:47:07,880 --> 00:47:08,080 THE 1316 00:47:08,080 --> 00:47:13,120 SAME TO HAVE NEURON-TYPE 1317 00:47:13,120 --> 00:47:13,760 SPECIFICITY DEPENDING ON THE 1318 00:47:13,760 --> 00:47:14,240 DOWNSTREAM NEURON. 1319 00:47:14,240 --> 00:47:17,080 --- 1320 00:47:17,080 --> 00:47:19,080 SO THIS IS ALL PUBLISHED 1321 00:47:19,080 --> 00:47:19,320 WORK, 1322 00:47:19,320 --> 00:47:19,920 BUT YOU STILL HAVE NOT ANSWER 1323 00:47:19,920 --> 00:47:20,120 THE 1324 00:47:20,120 --> 00:47:22,120 QUESTION EVEN IN C ELEGANS, 1325 00:47:22,120 --> 00:47:22,320 HOW 1326 00:47:22,320 --> 00:47:28,600 IS THE DLK LEVEL UPREGULATED? 1327 00:47:28,600 --> 00:47:30,440 SO I'M GOING TO ACTIVELY WORK 1328 00:47:30,440 --> 00:47:31,600 IN 1329 00:47:31,600 --> 00:47:33,320 C ELEGANS. 1330 00:47:33,320 --> 00:47:34,960 THIS IS A WORK FROM A 1331 00:47:34,960 --> 00:47:35,640 GRADUATE 1332 00:47:35,640 --> 00:47:39,040 STUDENT AND POSTDOC -- WHAT 1333 00:47:39,040 --> 00:47:39,280 THEY 1334 00:47:39,280 --> 00:47:41,400 DID IS THEY TOOK ADVANTAGE OF 1335 00:47:41,400 --> 00:47:45,680 GENETIC TAGGING IN DLK AND 1336 00:47:45,680 --> 00:47:48,160 DECIDED TO LOOK FOR WHAT 1337 00:47:48,160 --> 00:47:48,480 COULD BE 1338 00:47:48,480 --> 00:47:49,920 UPREGULATED. 1339 00:47:49,920 --> 00:47:51,320 SO WHAT WE ARE SHOWING HERE 1340 00:47:51,320 --> 00:47:51,520 IS 1341 00:47:51,520 --> 00:47:56,840 THE C ELEGANS WORM AND THE -- 1342 00:47:56,840 --> 00:47:57,240 IS 1343 00:47:57,240 --> 00:48:03,360 HERE WITH LESS SOMA AND THE 1344 00:48:03,360 --> 00:48:03,560 -- 1345 00:48:03,560 --> 00:48:07,160 CONNECTED HERE AND WOULD WE 1346 00:48:07,160 --> 00:48:07,840 SEE 1347 00:48:07,840 --> 00:48:13,440 ENDOGENOUS DLK, I CAN SEE ON 1348 00:48:13,440 --> 00:48:14,040 SCREEN, I DON'T KNOW IF IT 1349 00:48:14,040 --> 00:48:14,280 WILL 1350 00:48:14,280 --> 00:48:15,960 SHOW UP, THEN THEY FEED THE 1351 00:48:15,960 --> 00:48:16,200 WORM 1352 00:48:16,200 --> 00:48:19,080 WITH -- AND ISOLATED MUTANTS 1353 00:48:19,080 --> 00:48:19,320 THAT 1354 00:48:19,320 --> 00:48:22,040 WILL INCREASE THE DLK LEVEL, 1355 00:48:22,040 --> 00:48:22,840 LIKE 1356 00:48:22,840 --> 00:48:25,720 IN TRANSGENIC MICE, BUT WE 1357 00:48:25,720 --> 00:48:25,920 ARE 1358 00:48:25,920 --> 00:48:27,680 LOOKING FOR MUTATION. 1359 00:48:27,680 --> 00:48:28,760 SURE ENOUGH WE GOT A POSITIVE 1360 00:48:28,760 --> 00:48:33,960 CONTROL BECAUSE DLK NOT ONLY 1361 00:48:33,960 --> 00:48:36,720 INHIBITED DEGRADED THE LIGASE 1362 00:48:36,720 --> 00:48:41,040 (INDISCERNIBLE) -- ALSO 1363 00:48:41,040 --> 00:48:45,080 IDENTIFIED A TUBULIN, 1364 00:48:45,080 --> 00:48:46,240 UNEXPECTED AND IT IS CALLED 1365 00:48:46,240 --> 00:48:47,240 THE 1366 00:48:47,240 --> 00:48:52,200 BEN-1 AND THE C ELEGANS BEN-1 1367 00:48:52,200 --> 00:48:53,720 IS 1368 00:48:53,720 --> 00:48:54,200 QUITE WELL STUDIED, 1369 00:48:54,200 --> 00:48:55,360 STUDY FOR SOME YEARS FROM -- 1370 00:48:55,360 --> 00:48:58,320 GROUP. 1371 00:48:58,320 --> 00:49:06,560 IT IS A NEURON SPECIFIC B- 1372 00:49:06,560 --> 00:49:07,120 TUBULIN ISOLATED BECAUSE 1373 00:49:07,120 --> 00:49:07,360 WORMS 1374 00:49:07,360 --> 00:49:12,640 ARE SENSITIVE TO THIS CALLED 1375 00:49:12,640 --> 00:49:13,240 BENOMYL, AND EVENTUALLY TO 1376 00:49:13,240 --> 00:49:13,520 BECOME 1377 00:49:13,520 --> 00:49:14,320 STERILE AND THEY DIE. 1378 00:49:14,320 --> 00:49:19,000 LOSS OF FUNCTION OF BEN-1 IS 1379 00:49:19,000 --> 00:49:23,000 RESISTANT TO BENOMYL, AND 1380 00:49:23,000 --> 00:49:23,240 THEN 1381 00:49:23,240 --> 00:49:23,800 THEY GROW PERFECTLY FINE 1382 00:49:23,800 --> 00:49:24,080 WHEREAS 1383 00:49:24,080 --> 00:49:27,680 OUR MUTATION IS THIS ONE IN 1384 00:49:27,680 --> 00:49:29,480 THE 1385 00:49:29,480 --> 00:49:35,000 TUBULIN. TUBULIN'S ARE GTPA, 1386 00:49:35,000 --> 00:49:35,200 AND 1387 00:49:35,200 --> 00:49:37,960 THEY ARE PRETTY ACTIVE AND 1388 00:49:37,960 --> 00:49:38,200 THEY 1389 00:49:38,200 --> 00:49:38,880 FOLD EXTENSIVELY AND THERE IS 1390 00:49:38,880 --> 00:49:42,920 THIS LOOP CALLED THE T7 LOOP, 1391 00:49:42,920 --> 00:49:43,120 IN 1392 00:49:43,120 --> 00:49:48,520 THE ALPHA-BETA TUBLING 1393 00:49:48,520 --> 00:49:48,840 INTERFACE 1394 00:49:48,840 --> 00:49:51,840 IN OUR MUTATIONS AFFECTING 1395 00:49:51,840 --> 00:49:52,880 THIS 1396 00:49:52,880 --> 00:49:57,360 LOOP, IT BEHAVED -- 1397 00:49:57,360 --> 00:49:58,840 (INDISCERNIBLE)-- BECAUSE IT 1398 00:49:58,840 --> 00:49:59,040 IS 1399 00:49:59,040 --> 00:50:03,080 HYPERSENSITIVE TO BENOMYL. 1400 00:50:03,080 --> 00:50:07,120 THE GENE IS HARD TO REMEMBER 1401 00:50:07,120 --> 00:50:07,360 BUT 1402 00:50:07,360 --> 00:50:13,120 EASY TO MAKE FOR THOSE 1403 00:50:13,120 --> 00:50:13,400 FAMILIAR 1404 00:50:13,400 --> 00:50:19,040 WITH THE VERTEBRAE TUBULI. 1405 00:50:19,040 --> 00:50:19,560 THERE HAVE BEEN MANY 1406 00:50:19,560 --> 00:50:21,520 MUTATIONS, 1407 00:50:21,520 --> 00:50:30,080 DISRUPTING HUMAN TUBB3 1408 00:50:30,080 --> 00:50:30,360 RESULTING 1409 00:50:30,360 --> 00:50:32,680 IN DISORDERS. 1410 00:50:32,680 --> 00:50:35,840 BASICALLY THERE IS A LACK OF 1411 00:50:35,840 --> 00:50:38,600 MAMMALIAN TUBB3, BECAUSE IT 1412 00:50:38,600 --> 00:50:40,160 ALSO 1413 00:50:40,160 --> 00:50:44,120 CONVERS SENSITIVITY TO 1414 00:50:44,120 --> 00:50:44,840 BENOMYL. 1415 00:50:44,840 --> 00:50:49,520 --- 1416 00:50:49,520 --> 00:50:50,120 THIS MUTATION THAT I SHOWED 1417 00:50:50,120 --> 00:50:50,320 YOU 1418 00:50:50,320 --> 00:50:51,000 IS ISOLATED BECAUSE IT 1419 00:50:51,000 --> 00:50:51,880 ISOLATED 1420 00:50:51,880 --> 00:50:55,640 THE DLK LEVEL, AND NOT ONLY 1421 00:50:55,640 --> 00:50:55,960 IT UP 1422 00:50:55,960 --> 00:51:00,240 REGULATES BUT IT ACTIVATES 1423 00:51:00,240 --> 00:51:00,440 ITS 1424 00:51:00,440 --> 00:51:00,920 DOWNSTREAM SIGNALING. 1425 00:51:00,920 --> 00:51:06,480 DOWNSTREAM OF THE DLK IS A 1426 00:51:06,480 --> 00:51:10,280 PRESCRIPTION FACTOR CEBP-1. 1427 00:51:10,280 --> 00:51:13,880 IT UBIQUITOUSLY IS EXPRESSING 1428 00:51:13,880 --> 00:51:14,080 IN 1429 00:51:14,080 --> 00:51:14,720 ALL TISSUES, VERY LOW IN THE 1430 00:51:14,720 --> 00:51:18,120 NEURONS IN THE HEAD AND IN 1431 00:51:18,120 --> 00:51:18,920 MOST 1432 00:51:18,920 --> 00:51:21,400 PLACES. 1433 00:51:21,400 --> 00:51:22,040 BUT IN THE MUTANT YOU CAN SEE 1434 00:51:22,040 --> 00:51:23,000 IT'S EXPRESSION LEVELS 1435 00:51:23,000 --> 00:51:28,160 UPREGULATED IN MOST NEURONS, 1436 00:51:28,160 --> 00:51:29,360 NOT 1437 00:51:29,360 --> 00:51:31,080 IN NON-NEURAL TISSUES. 1438 00:51:31,080 --> 00:51:34,440 AND THIS MUTATION IS ACTING 1439 00:51:34,440 --> 00:51:34,640 IN 1440 00:51:34,640 --> 00:51:42,920 PARALLEL TO UP REGULATE LOSS 1441 00:51:42,920 --> 00:51:44,280 OF 1442 00:51:44,280 --> 00:51:47,800 FUNCTION WHERE THE DUCKTAPE 1443 00:51:47,800 --> 00:51:48,160 WORM, 1444 00:51:48,160 --> 00:51:51,360 WHICH MOVES WELL, 1445 00:51:51,360 --> 00:51:57,360 THERE IS A LOSS OF FUNCTION, 1446 00:51:57,360 --> 00:51:57,600 ALSO 1447 00:51:57,600 --> 00:51:58,200 PRETTY MUCH NORMAL BUT DOUBLE 1448 00:51:58,200 --> 00:51:59,120 MUTANTS ARE PARALYZED. 1449 00:51:59,120 --> 00:51:59,640 AND WHEN WE LOOK AT THE 1450 00:51:59,640 --> 00:52:02,800 SYNOPSIS, 1451 00:52:02,800 --> 00:52:03,360 YOU CAN SEE THE SYNOPTIC 1452 00:52:03,360 --> 00:52:03,640 PUNCTURE 1453 00:52:03,640 --> 00:52:07,760 IN THE WILD TYPE, 1454 00:52:07,760 --> 00:52:10,120 VERY DISCREET. 1455 00:52:10,120 --> 00:52:14,200 THE BEN-1 FUNCTION IS ALSO 1456 00:52:14,200 --> 00:52:14,760 DISCRETE, 1457 00:52:14,760 --> 00:52:16,640 DISRUPTING THE SHAPE OF THE 1458 00:52:16,640 --> 00:52:18,320 SYNOPSIS AND THE DOUBLE 1459 00:52:18,320 --> 00:52:18,920 MUTANTS 1460 00:52:18,920 --> 00:52:21,560 LOOSE BASICALLY MOST SYNAPSES 1461 00:52:21,560 --> 00:52:21,760 AND 1462 00:52:21,760 --> 00:52:22,360 MOST ARE PARALYZED. 1463 00:52:22,360 --> 00:52:28,600 SO THE BEN-1 FUNCTIONS ACT IN 1464 00:52:28,600 --> 00:52:30,920 A 1465 00:52:30,920 --> 00:52:33,360 PARALLEL PATHWAY, IT IS A 1466 00:52:33,360 --> 00:52:34,640 TUBULIN, AND WE WERE USING 1467 00:52:34,640 --> 00:52:38,360 THE 1468 00:52:38,360 --> 00:52:42,320 EBB -BINDING ASSAY. 1469 00:52:42,320 --> 00:52:44,600 IN THIS PARTICULAR FUNCTION 1470 00:52:44,600 --> 00:52:47,320 ACTUALLY, THE MICROTUBULE 1471 00:52:47,320 --> 00:52:54,800 DYNAMICS ARE ONLY SUBTLY 1472 00:52:54,800 --> 00:52:55,120 ALTERED, 1473 00:52:55,120 --> 00:52:55,480 NOT DRAMATIC. 1474 00:52:55,480 --> 00:52:59,080 THIS FUNCTION CHANGE THE 1475 00:52:59,080 --> 00:52:59,680 MICROTUBULE DYNAMICS AND IT 1476 00:52:59,680 --> 00:53:02,800 ACTIVATES THE DLK MODERATELY. 1477 00:53:02,800 --> 00:53:05,000 IT HAS A PROTECTIVE ROLE IN 1478 00:53:05,000 --> 00:53:08,120 SYNOPSIS IN MULTI NEURAL 1479 00:53:08,120 --> 00:53:10,120 FUNCTION. 1480 00:53:10,120 --> 00:53:11,280 THIS SHOWS LOOKING AT THE 1481 00:53:11,280 --> 00:53:13,000 SYNAPTIC PUNCTURE, AND 1482 00:53:13,000 --> 00:53:13,320 LOOKING AT 1483 00:53:13,320 --> 00:53:18,640 THE -- FUNCTION AND DLK LOSS 1484 00:53:18,640 --> 00:53:19,560 FUNCTION, ALL LOOK FAIRLY 1485 00:53:19,560 --> 00:53:19,840 SIMILAR 1486 00:53:19,840 --> 00:53:23,960 BUT THE REASON WE SAY 1487 00:53:23,960 --> 00:53:25,720 ACTIVATED 1488 00:53:25,720 --> 00:53:28,280 DLK FUNCTION PROTECTS 1489 00:53:28,280 --> 00:53:28,840 SYNAPSES IS 1490 00:53:28,840 --> 00:53:34,400 WE TAKE OUT THE DLK IN THE 1491 00:53:34,400 --> 00:53:34,640 BEN-1 1492 00:53:34,640 --> 00:53:35,280 IN THE FUNCTIONAL MUTANT AND 1493 00:53:35,280 --> 00:53:35,480 THE 1494 00:53:35,480 --> 00:53:37,880 SYNOPSIS GETS SEVERELY 1495 00:53:37,880 --> 00:53:38,480 DISRUPTED. 1496 00:53:38,480 --> 00:53:39,080 THIS IS OPPOSITE BY SAYING 1497 00:53:39,080 --> 00:53:39,320 THAT 1498 00:53:39,320 --> 00:53:46,480 WHEN WE DID -- LOSS OF 1499 00:53:46,480 --> 00:53:46,920 FUNCTION, 1500 00:53:46,920 --> 00:53:50,880 WHEN WE TOOK OUT DLK, IT 1501 00:53:50,880 --> 00:53:51,160 RESTORED 1502 00:53:51,160 --> 00:53:51,760 FUNCTIONS BUT HERE WE TOOK 1503 00:53:51,760 --> 00:53:51,960 OUT 1504 00:53:51,960 --> 00:53:55,800 THE DLK AND MADE THE SYNAPSES 1505 00:53:55,800 --> 00:53:56,480 WORSE, AND THE MOTOR BEHAVIOR 1506 00:53:56,480 --> 00:53:58,080 WORSE. 1507 00:53:58,080 --> 00:53:58,800 THIS IS A MODERATE INCREASE 1508 00:53:58,800 --> 00:53:59,160 OF 1509 00:53:59,160 --> 00:54:00,640 DLK LEVEL. 1510 00:54:00,640 --> 00:54:03,520 --- 1511 00:54:03,520 --> 00:54:06,800 WHEN WE TREAT THE ANIMALS 1512 00:54:06,800 --> 00:54:16,960 WITH 1513 00:54:17,760 --> 00:54:18,440 BENOMYL, IT DEPOLARIZES MANY 1514 00:54:18,440 --> 00:54:22,800 TUBULINS. 1515 00:54:22,800 --> 00:54:23,440 IT IS SEVERELY ALTERED BOTH 1516 00:54:23,440 --> 00:54:23,640 IN 1517 00:54:23,640 --> 00:54:24,280 THE NUMBER OF TRACKS AS WELL 1518 00:54:24,280 --> 00:54:24,480 AS 1519 00:54:24,480 --> 00:54:24,880 SHORT NUMBERS OF 1520 00:54:24,880 --> 00:54:25,680 (INDISCERNIBLE). 1521 00:54:25,680 --> 00:54:26,920 --- 1522 00:54:26,920 --> 00:54:28,120 IN THIS CASE WHAT WE SEE IS 1523 00:54:28,120 --> 00:54:29,360 THAT 1524 00:54:29,360 --> 00:54:39,040 BENOMYL TREATMENT DEPORALIZES 1525 00:54:39,040 --> 00:54:39,240 THE 1526 00:54:39,240 --> 00:54:39,760 MAJORITY OF THE BUCKET 1527 00:54:39,760 --> 00:54:40,040 TUBULES. 1528 00:54:40,040 --> 00:54:50,560 THIS IS -- THE ANIMALS WERE 1529 00:54:51,280 --> 00:54:54,160 PARALYZED AND DLK BLOCKS THIS 1530 00:54:54,160 --> 00:54:55,640 EFFECT. 1531 00:54:55,640 --> 00:55:01,240 IN THE SEVERE CASE, DLK IS 1532 00:55:01,240 --> 00:55:09,280 BASICALLY BLOCKING THE 1533 00:55:09,280 --> 00:55:09,880 MICROTUBULE DISRUPTION AND 1534 00:55:09,880 --> 00:55:11,320 INDUCED A SYNAPTIC DISRUPTION 1535 00:55:11,320 --> 00:55:11,520 AND 1536 00:55:11,520 --> 00:55:12,400 IN THIS CASE, WE SEE A DOSAGE 1537 00:55:12,400 --> 00:55:15,000 EFFECT. 1538 00:55:15,000 --> 00:55:15,480 MODEST DISRUPTION OF 1539 00:55:15,480 --> 00:55:16,880 MICROTUBULES 1540 00:55:16,880 --> 00:55:18,680 -- THIS IS MY LAST SLIDE -- 1541 00:55:18,680 --> 00:55:18,920 IN 1542 00:55:18,920 --> 00:55:20,920 THE STEADY-STATE DLK 1543 00:55:20,920 --> 00:55:25,240 SIGNALING IS 1544 00:55:25,240 --> 00:55:25,840 MODERATELY ACTIVITY TO CREATE 1545 00:55:25,840 --> 00:55:26,040 A 1546 00:55:26,040 --> 00:55:29,640 TIGHTLY REGULATED -- LIGASE 1547 00:55:29,640 --> 00:55:29,840 OF 1548 00:55:29,840 --> 00:55:30,760 THE SIGNALING CONSTANTLY IS 1549 00:55:30,760 --> 00:55:31,000 UNDER 1550 00:55:31,000 --> 00:55:31,880 CONTROL. 1551 00:55:31,880 --> 00:55:37,640 WITH THE -- FUNCTION THERE IS 1552 00:55:37,640 --> 00:55:37,800 A 1553 00:55:37,800 --> 00:55:42,960 SLIGHT CHANGE IN MICROTUBULE 1554 00:55:42,960 --> 00:55:43,560 DYNAMICS WHICH UP REGULATES 1555 00:55:43,560 --> 00:55:43,760 THIS 1556 00:55:43,760 --> 00:55:47,200 SLIGHTLY AND IT ALTERS THIS. 1557 00:55:47,200 --> 00:55:49,160 AND THE PURPOSE OF OPERATING 1558 00:55:49,160 --> 00:55:49,360 THE 1559 00:55:49,360 --> 00:55:54,920 SIGNALING IS THE CHANGES OF 1560 00:55:54,920 --> 00:55:58,400 MICROTUBULE DYNAMICS TO KEEP 1561 00:55:58,400 --> 00:55:59,560 THE 1562 00:55:59,560 --> 00:56:01,200 SYNAPSES BACK TO NORMAL IN 1563 00:56:01,200 --> 00:56:01,440 THIS 1564 00:56:01,440 --> 00:56:05,200 CASE DLK IS UPREGULATED, AND 1565 00:56:05,200 --> 00:56:06,920 IN 1566 00:56:06,920 --> 00:56:10,920 THIS CASE IT EXCEEDED THE 1567 00:56:10,920 --> 00:56:11,160 FIRST 1568 00:56:11,160 --> 00:56:11,920 THRESHOLD AND SYNOPSES BECOME 1569 00:56:11,920 --> 00:56:12,920 TOTALLY UNSTABLE AND NEURONS 1570 00:56:12,920 --> 00:56:13,120 NO 1571 00:56:13,120 --> 00:56:14,400 LONGER FUNCTION NORMALLY. 1572 00:56:14,400 --> 00:56:16,960 SO THAT LEAVES ME WITH MY 1573 00:56:16,960 --> 00:56:17,760 WORKING 1574 00:56:17,760 --> 00:56:24,280 MODEL FROM STARTING FROM ONE 1575 00:56:24,280 --> 00:56:25,840 SCREEN TO ANOTHER SCREEN 1576 00:56:25,840 --> 00:56:26,120 TESTING 1577 00:56:26,120 --> 00:56:27,520 OUR HYPOTHESES USING MOUSE 1578 00:56:27,520 --> 00:56:29,840 MODELS. 1579 00:56:29,840 --> 00:56:30,720 WE LIKE TO THINK THAT THIS 1580 00:56:30,720 --> 00:56:31,000 STRESS 1581 00:56:31,000 --> 00:56:37,200 KINASE DLK IS ACTIVATION, 1582 00:56:37,200 --> 00:56:37,880 TIGHTLY 1583 00:56:37,880 --> 00:56:38,640 COUPLED TO MICROTUBULE 1584 00:56:38,640 --> 00:56:38,920 DYNAMICS, 1585 00:56:38,920 --> 00:56:40,960 NEURON SPECIFIC TO AN ISO 1586 00:56:40,960 --> 00:56:43,840 FORM. 1587 00:56:43,840 --> 00:56:45,800 AND HOW THIS TIGHT COUPLING 1588 00:56:45,800 --> 00:56:48,560 IS 1589 00:56:48,560 --> 00:56:57,640 ACTUALLY MECHANISTICALLY, AT 1590 00:56:57,640 --> 00:56:57,880 WHAT 1591 00:56:57,880 --> 00:56:58,520 LEVEL IS A QUESITON MARK THAT 1592 00:56:58,520 --> 00:56:58,720 WE 1593 00:56:58,720 --> 00:57:01,640 DO NOT KNOW. 1594 00:57:01,640 --> 00:57:02,200 IT WILL CREATE A CONTEXT 1595 00:57:02,200 --> 00:57:02,800 -DEPENDENT OUTCOME WHETHER 1596 00:57:02,800 --> 00:57:04,120 NEURONS WILL SURVIVE, 1597 00:57:04,120 --> 00:57:06,640 FORM NORMAL SYNOPSES, 1598 00:57:06,640 --> 00:57:12,960 REGENERATING OR DEGENERATING. 1599 00:57:12,960 --> 00:57:13,960 WE HAVE A LOT MORE WORK TO DO 1600 00:57:13,960 --> 00:57:14,160 BUT 1601 00:57:14,160 --> 00:57:18,920 I WANT TO END THE TALK 1602 00:57:18,920 --> 00:57:19,200 SHOWING 1603 00:57:19,200 --> 00:57:19,880 THE PEOPLE IN MY LAB. 1604 00:57:19,880 --> 00:57:20,480 AND THE FIRST OUTINGS LAST 1605 00:57:20,480 --> 00:57:20,720 YEAR 1606 00:57:20,720 --> 00:57:21,320 AFTER THE PANDEMIC AND THE 1607 00:57:21,320 --> 00:57:21,520 KEY 1608 00:57:21,520 --> 00:57:21,760 PEOPLE 1609 00:57:21,760 --> 00:57:24,480 WHO DID THE WORK ARE ERIN, 1610 00:57:24,480 --> 00:57:29,840 RICHIE, -- MY COLLABORATOR 1611 00:57:29,840 --> 00:57:30,040 AT 1612 00:57:30,040 --> 00:57:33,240 UCSD -- AND THE C ELEGANS 1613 00:57:33,240 --> 00:57:38,000 AXON 1614 00:57:38,000 --> 00:57:39,080 AND COLLABORATION IN SPINAL 1615 00:57:39,080 --> 00:57:40,000 CORD 1616 00:57:40,000 --> 00:57:43,680 INJURY, AND THEY ALL 1617 00:57:43,680 --> 00:57:44,040 CONTRIBUTED 1618 00:57:44,040 --> 00:57:44,800 TO EACH OF THE STORIES THAT I 1619 00:57:44,800 --> 00:57:47,040 TALKED TO YOU ABOUT. THANK 1620 00:57:47,040 --> 00:57:50,840 YOU TO 1621 00:57:50,840 --> 00:57:52,480 NIH FOR A VERY LONG SUSTAIN 1622 00:57:52,480 --> 00:57:55,000 SUPPORT, AND IT HELP ME TO 1623 00:57:55,000 --> 00:57:55,720 SET UP 1624 00:57:55,720 --> 00:58:01,640 THE MOUSE WORK AND JUNOR 1625 00:58:01,640 --> 00:58:01,920 SEAU, A 1626 00:58:01,920 --> 00:58:03,600 HERO IN THE FOOTBALL ERA FOR 1627 00:58:03,600 --> 00:58:04,760 WHICH WE HAVE THE HONOR TO BE 1628 00:58:04,760 --> 00:58:08,880 THE 1629 00:58:08,880 --> 00:58:13,040 CHAIR AND THE REST OF THE 1630 00:58:13,040 --> 00:58:13,360 SUPPLIES. 1631 00:58:13,360 --> 00:58:14,000 THANK YOU FOR COMING. 1632 00:58:14,000 --> 00:58:19,840 >> [APPLAUSE] 1633 00:58:19,840 --> 00:58:20,480 >> ANYONE IS FREE TO USE THE 1634 00:58:20,480 --> 00:58:20,960 MICROPHONES TO ASK A 1635 00:58:20,960 --> 00:58:23,280 QUESTION. 1636 00:58:23,280 --> 00:58:25,680 I HAVE ONE THAT HAS COME IN 1637 00:58:25,680 --> 00:58:25,920 ONLINE. 1638 00:58:25,920 --> 00:58:27,080 A TWO-PART QUESTION. 1639 00:58:27,080 --> 00:58:31,120 THIS IS FROM -- A RESEARCHER 1640 00:58:31,120 --> 00:58:33,840 HERE. 1641 00:58:33,840 --> 00:58:34,480 THANK YOU SO MUCH FOR SHARING 1642 00:58:34,480 --> 00:58:37,640 THIS AWESOME WORK, VERY 1643 00:58:37,640 --> 00:58:38,000 INTERESTING. 1644 00:58:38,000 --> 00:58:39,040 YOU FOCUSED ON INJURY INDUCED 1645 00:58:39,040 --> 00:58:42,480 REGENERATION IN AXONS. 1646 00:58:42,480 --> 00:58:43,440 IS THE SAME MECHANISM 1647 00:58:43,440 --> 00:58:43,880 EMPLOYED 1648 00:58:43,880 --> 00:58:46,840 FOR DAMAGED DENDRITES? 1649 00:58:46,840 --> 00:58:50,080 PART TWO, 1650 00:58:50,080 --> 00:58:50,640 DOES THE INJURY INDUCED 1651 00:58:50,640 --> 00:58:52,840 REGENERATION FULLY RESTORE 1652 00:58:52,840 --> 00:58:55,920 FUNCTIONS OF THE DAMAGED AXON 1653 00:58:55,920 --> 00:58:56,120 TO 1654 00:58:56,120 --> 00:58:59,320 THE STATE BEFORE THE INJURY? 1655 00:58:59,320 --> 00:59:00,400 AXONS CARRY MANY FUNCTIONAL 1656 00:59:00,400 --> 00:59:05,520 UNITS, FOR INSTANCE AXON 1657 00:59:05,520 --> 00:59:08,600 INITIATED SEGMENT- AND 1658 00:59:08,600 --> 00:59:10,440 (INDISCERNIBLE) AND ACQUIRED 1659 00:59:10,440 --> 00:59:11,160 SYNOPTIC STRENGTH. 1660 00:59:11,160 --> 00:59:11,760 THANK YOU FOR ANSWERING THE 1661 00:59:11,760 --> 00:59:12,040 QUESTION. 1662 00:59:12,040 --> 00:59:14,000 >> DR. JIN: 1663 00:59:14,000 --> 00:59:16,160 THANK YOU FOR THE QUESTION. 1664 00:59:16,160 --> 00:59:18,040 THE FIRST IS EASY TO ANSWER. 1665 00:59:18,040 --> 00:59:20,640 I DID FOCUS IN ALL OF MY WORK 1666 00:59:20,640 --> 00:59:22,720 ON 1667 00:59:22,720 --> 00:59:23,280 AXON REGENERATION BUT ONE 1668 00:59:23,280 --> 00:59:23,520 THING 1669 00:59:23,520 --> 00:59:24,280 TO KEEP IN MIND IS THAT C 1670 00:59:24,280 --> 00:59:25,280 ELEGANS 1671 00:59:25,280 --> 00:59:28,520 ARE SIMPLE AXONS, AND SOME OF 1672 00:59:28,520 --> 00:59:29,400 THEM ARE MIXED POLARITY. 1673 00:59:29,400 --> 00:59:32,200 IN OUR LAB, WE WERE WORKING 1674 00:59:32,200 --> 00:59:32,440 WITH 1675 00:59:32,440 --> 00:59:38,160 THE C ELEGANS TESTING 1676 00:59:38,160 --> 00:59:38,440 MULTIPLE 1677 00:59:38,440 --> 00:59:42,120 TYPE OF NEURIDES, AND PURITY 1678 00:59:42,120 --> 00:59:44,200 DENDRITE AND GENERALLY C 1679 00:59:44,200 --> 00:59:45,520 ELEGANS 1680 00:59:45,520 --> 00:59:46,840 DENDRITES DO NOT GENERATE NOT 1681 00:59:46,840 --> 00:59:49,600 BECAUSE THE DLK IS NOT THERE. 1682 00:59:49,600 --> 00:59:55,240 THEY DON'T REGENERATE. 1683 00:59:55,240 --> 01:00:02,120 I WILL TALK ABOUT AN OFFER -- 1684 01:00:02,120 --> 01:00:04,560 THE 1685 01:00:04,560 --> 01:00:07,320 INTRA CEPHULA (PHONETIC) 1686 01:00:07,320 --> 01:00:11,000 (INDISCERNIBLE) -- 1687 01:00:11,000 --> 01:00:13,360 OTHER PEOPLE'S WORK. 1688 01:00:13,360 --> 01:00:14,120 IN THAT SENSE I WILL SIMPLY 1689 01:00:14,120 --> 01:00:14,320 SAY 1690 01:00:14,320 --> 01:00:16,880 THAT BECAUSE C ELEGANS AXONS 1691 01:00:16,880 --> 01:00:17,920 HAVE 1692 01:00:17,920 --> 01:00:20,680 A UNIQUE ARCHITECTURE WE 1693 01:00:20,680 --> 01:00:21,000 FOCUS ON 1694 01:00:21,000 --> 01:00:21,440 AXON REGENERATION. 1695 01:00:21,440 --> 01:00:22,280 --- 1696 01:00:22,280 --> 01:00:26,920 THE SECOND QUESTION, AXON 1697 01:00:26,920 --> 01:00:29,080 REGENERATION IS VERY COMPLEX, 1698 01:00:29,080 --> 01:00:29,280 I 1699 01:00:29,280 --> 01:00:29,640 TOTALLY AGREE. 1700 01:00:29,640 --> 01:00:31,680 AT THIS STAGE FROM THE C 1701 01:00:31,680 --> 01:00:34,280 ELEGANS 1702 01:00:34,280 --> 01:00:38,280 WORK, OUR GOAL IS TO LOOK FOR 1703 01:00:38,280 --> 01:00:40,400 GENES THAT WILL RESPOND TO 1704 01:00:40,400 --> 01:00:41,040 INJURY, AND IN THAT SENSE WE 1705 01:00:41,040 --> 01:00:41,280 HAVE 1706 01:00:41,280 --> 01:00:44,720 NOT EXPLORED INTO IN-DEPTH OF 1707 01:00:44,720 --> 01:00:48,080 DIFFERENT FORMS WHETHER THE 1708 01:00:48,080 --> 01:00:48,320 AXON 1709 01:00:48,320 --> 01:00:54,920 WILL SPROUT OR BRANCH AND 1710 01:00:54,920 --> 01:00:55,160 FORM 1711 01:00:55,160 --> 01:00:55,760 SYNOPSES IN ANY WAY BUT IN 1712 01:00:55,760 --> 01:00:57,600 COLLABORATION WITH -- LAB, WE 1713 01:00:57,600 --> 01:01:01,840 LOOKED INTO SPINAL CORD 1714 01:01:01,840 --> 01:01:02,080 INJURY 1715 01:01:02,080 --> 01:01:08,520 WHICH HAS WHEN THEY DO DORSAL 1716 01:01:08,520 --> 01:01:10,520 HEMI TRANSECTION, THE SPINAL 1717 01:01:10,520 --> 01:01:10,720 CORD 1718 01:01:10,720 --> 01:01:15,800 COULD HAVE A SPARED AXON AND 1719 01:01:15,800 --> 01:01:16,600 SPROUTING, AND THEY CAN ALSO 1720 01:01:16,600 --> 01:01:17,560 PULL 1721 01:01:17,560 --> 01:01:20,360 IN AN INJURED AXON ALSO 1722 01:01:20,360 --> 01:01:20,960 SPROUTING 1723 01:01:20,960 --> 01:01:26,040 AND WHAT -- GROUP HAS 1724 01:01:26,040 --> 01:01:26,360 OBSERVED 1725 01:01:26,360 --> 01:01:28,280 THAT BOTH EFFECTS ARE 1726 01:01:28,280 --> 01:01:29,680 DEPENDENT 1727 01:01:29,680 --> 01:01:32,080 ON DLK. 1728 01:01:32,080 --> 01:01:38,080 SO THERE ARE SINGLING EVENTS 1729 01:01:38,080 --> 01:01:38,280 THAT 1730 01:01:38,280 --> 01:01:38,920 SOMEHOW THE NEURON WILL SENSE 1731 01:01:38,920 --> 01:01:39,120 THE 1732 01:01:39,120 --> 01:01:39,680 INJURY PRESENT AND WILL 1733 01:01:39,680 --> 01:01:41,560 RESPONSE. 1734 01:01:41,560 --> 01:01:43,000 WE HAVE NOT SEEN THE 1735 01:01:43,000 --> 01:01:43,320 REFORMING 1736 01:01:43,320 --> 01:01:44,280 SYNOPSES BY MANIPULATIVE 1737 01:01:44,280 --> 01:01:44,560 SINGLE 1738 01:01:44,560 --> 01:01:46,320 GENETIC FACTORS, 1739 01:01:46,320 --> 01:01:47,760 SOMETIMES USING MULTIPLE 1740 01:01:47,760 --> 01:01:49,920 FACTORS 1741 01:01:49,920 --> 01:01:56,560 CAN MAKE THE INJURED AXONS 1742 01:01:56,560 --> 01:01:56,800 GROW 1743 01:01:56,800 --> 01:01:57,400 BETTER BUT THE SYNOPSES IS 1744 01:01:57,400 --> 01:01:57,600 NOT 1745 01:01:57,600 --> 01:01:58,000 FORMED PROPERLY. 1746 01:01:58,000 --> 01:01:58,200 --- 1747 01:01:58,200 --> 01:02:02,120 IN C ELEGANS, SOME OF THE 1748 01:02:02,120 --> 01:02:02,400 WORK WE 1749 01:02:02,400 --> 01:02:04,440 KNOW IS THAT THEY CAN BE 1750 01:02:04,440 --> 01:02:06,560 DISCONNECTED AND HAVE A 1751 01:02:06,560 --> 01:02:06,840 CERTAIN 1752 01:02:06,840 --> 01:02:07,440 DEGRADED FUNCTION DEPENDING 1753 01:02:07,440 --> 01:02:07,640 ON 1754 01:02:07,640 --> 01:02:08,280 THE CIRCUITS, DEPENDING OF 1755 01:02:08,280 --> 01:02:09,600 THE 1756 01:02:09,600 --> 01:02:12,160 ULTIMATE SYNOPSES IS IN THE 1757 01:02:12,160 --> 01:02:14,400 (INDISCERNIBLE) MUSCLES. 1758 01:02:14,400 --> 01:02:17,520 >> THANK YOU VERY MUCH. 1759 01:02:17,520 --> 01:02:20,040 IT'S BEEN QUITE AN AMOUNT OF 1760 01:02:20,040 --> 01:02:22,240 INTERESTING STORIES. 1761 01:02:22,240 --> 01:02:25,320 MY QUESTION HAS TO DO WITH 1762 01:02:25,320 --> 01:02:25,520 THE 1763 01:02:25,520 --> 01:02:30,360 PREOWNED DOMAIN OF THE GRR 1764 01:02:30,360 --> 01:02:30,960 REQUIRED FOR BOTH FUNCTION 1765 01:02:30,960 --> 01:02:31,800 AND 1766 01:02:31,800 --> 01:02:33,840 DROPLET DISTRIBUTION. 1767 01:02:33,840 --> 01:02:42,920 YOU DESCRIBED THE 1768 01:02:42,920 --> 01:02:43,320 PHOSPHORYLATION 1769 01:02:43,320 --> 01:02:43,960 OF THAT DOMAINS WHICH IN MY 1770 01:02:43,960 --> 01:02:44,200 MIND 1771 01:02:44,200 --> 01:02:47,720 IS PRONE TO AGGREGATION. 1772 01:02:47,720 --> 01:02:53,800 AND SOMEBODY ELSE'S GROUP, 1773 01:02:53,800 --> 01:02:54,440 PHOSPHORYLATION ENHANCED THE 1774 01:02:54,440 --> 01:02:54,760 PARTITION. 1775 01:02:54,760 --> 01:02:55,320 ISN'T IT COUNTERINTUITIVE? 1776 01:02:55,320 --> 01:02:56,120 >> DR. JIN: 1777 01:02:56,120 --> 01:03:00,920 YEAH. 1778 01:03:00,920 --> 01:03:01,520 PHOSPHORYLATION, HOW IT IS 1779 01:03:01,520 --> 01:03:06,440 MODULATING AND SOMETIMES IT 1780 01:03:06,440 --> 01:03:06,680 COULD 1781 01:03:06,680 --> 01:03:10,760 BE TRIGGERING SIGNALS, THE 1782 01:03:10,760 --> 01:03:11,400 PHOSPHORYLATION ALSO CHANGES 1783 01:03:11,400 --> 01:03:11,600 THE 1784 01:03:11,600 --> 01:03:13,520 CHARGE ENVIRONMENT. 1785 01:03:13,520 --> 01:03:16,120 SO THE PHASE SEPARATION HAS A 1786 01:03:16,120 --> 01:03:16,320 LOT 1787 01:03:16,320 --> 01:03:17,720 TO DO WITH CROWDING, HOW YOU 1788 01:03:17,720 --> 01:03:20,680 GET 1789 01:03:20,680 --> 01:03:23,920 TO YOUR NEIGHBORS. 1790 01:03:23,920 --> 01:03:33,080 I HAVE SEEN SOMETIMES 1791 01:03:33,080 --> 01:03:33,360 REPORTING 1792 01:03:33,360 --> 01:03:35,080 THAT PHOSPHORYLATION PROMOTES 1793 01:03:35,080 --> 01:03:36,560 DROPLET FORMATION. 1794 01:03:36,560 --> 01:03:40,120 AND THE DROP TO SOLID-STATE. 1795 01:03:40,120 --> 01:03:42,920 A LOT OF THIS IS COMING, 1796 01:03:42,920 --> 01:03:43,720 EVENTUALLY DISCOVERING WHO IS 1797 01:03:43,720 --> 01:03:45,320 DROPLETS FORMING WITH. 1798 01:03:45,320 --> 01:03:47,120 >> I SEE. 1799 01:03:47,120 --> 01:03:49,200 >> DR. JIN: 1800 01:03:49,200 --> 01:03:52,040 WE HAVE NOT ACTUALLY IDENTIFY 1801 01:03:52,040 --> 01:03:56,720 THE 1802 01:03:56,720 --> 01:03:57,600 KINASE, EXCEPT WE HAVE 1803 01:03:57,600 --> 01:03:57,880 MUTATED 1804 01:03:57,880 --> 01:03:58,400 THE RESIDUES AND WE SEE 1805 01:03:58,400 --> 01:03:59,920 CHANGES 1806 01:03:59,920 --> 01:04:02,440 IN (INDISCERNIBLE). 1807 01:04:02,440 --> 01:04:07,440 >> THANK YOU. 1808 01:04:07,440 --> 01:04:12,600 >> CONCLUDING COMMENTS? 1809 01:04:12,600 --> 01:04:15,480 >> THANK YOU SO MUCH. 1810 01:04:15,480 --> 01:04:17,480 IT'S BEEN A PLEASURE TO SEE 1811 01:04:17,480 --> 01:04:17,680 ALL 1812 01:04:17,680 --> 01:04:19,600 OF THIS TOGETHER. 1813 01:04:19,600 --> 01:04:22,120 WE ALL HAVE BEEN READING YOUR 1814 01:04:22,120 --> 01:04:23,040 PAPERS, BUT SEEING HOW THEY 1815 01:04:23,040 --> 01:04:23,600 COME 1816 01:04:23,600 --> 01:04:24,080 TOGETHER IS QUITE 1817 01:04:24,080 --> 01:04:25,480 ENLIGHTENING. 1818 01:04:25,480 --> 01:04:30,680 AND I'M ALWAYS THRILLED TO 1819 01:04:30,680 --> 01:04:30,920 HEAR 1820 01:04:30,920 --> 01:04:33,320 YOUR STORIES. 1821 01:04:33,320 --> 01:04:38,240 AND I AM SURE MANY IN THE 1822 01:04:38,240 --> 01:04:39,080 AUDIENCE FEEL THE SAME. 1823 01:04:39,080 --> 01:04:39,800 THANK YOU. 1824 01:04:39,800 --> 01:04:41,880 >>DR. JIN: 1825 01:04:41,880 --> 01:04:42,000 THANK YOU VERY MUCH FOR YOUR 1826 01:04:42,000 --> 00:00:00,000 QUESTIONS.