1 00:00:06,659 --> 00:00:08,894 >> GOOD AFTERNOON, EVERYONE. MY 2 00:00:08,894 --> 00:00:12,031 NAME IS CHRIS ALEXANDER THE 3 00:00:12,031 --> 00:00:13,332 ASSOCIATE SCIENTIFIC DIRECTOR 4 00:00:13,332 --> 00:00:21,073 FOR NIHLBI DIR AND -- CHIEF OF 5 00:00:21,073 --> 00:00:27,212 THE LABORATORY STRUCTURAL 6 00:00:27,212 --> 00:00:27,813 CELLOL 7 00:00:27,813 --> 00:00:30,316 BIOLOGY -- TOKYO IN 1999 AND 8 00:00:30,316 --> 00:00:34,219 RECEIVED PHD IN BIOPHYSICS FROM 9 00:00:34,219 --> 00:00:36,021 UNIVERSITY OF TOKYO AND 10 00:00:36,021 --> 00:00:38,223 SOUTHWEST MEDICAL CENTER IN 2005 11 00:00:38,223 --> 00:00:42,328 AND PRIOR TO COMING TO NIH WAS 12 00:00:42,328 --> 00:00:44,964 AN INDEPENDENT GROUP LEADER IN 13 00:00:44,964 --> 00:00:47,266 CHEMISTRY IN GERMANY BEFORE 14 00:00:47,266 --> 00:00:51,437 MOVING HERE TO NIH DURING THE 15 00:00:51,437 --> 00:00:56,041 PANDE 16 00:00:56,041 --> 00:00:56,475 PANDEMIC. 17 00:00:56,475 --> 00:01:01,046 KRIO EM IS A CUTTING EDGE 18 00:01:01,046 --> 00:01:03,349 TECHNIQUE THAT BRIDGES 19 00:01:03,349 --> 00:01:07,620 RESOLUTION GAP BETWEEN LIGHT 20 00:01:07,620 --> 00:01:12,024 MICROSCOPY AND -- LAB USES 21 00:01:12,024 --> 00:01:16,729 INSITUCIO ELECTRO-TOMO GRAPHY 22 00:01:16,729 --> 00:01:21,333 AND CRYSTALO GRAPH IN VITRO 23 00:01:21,333 --> 00:01:23,235 INSTITUTION AND LIGHT MICROSCOPY 24 00:01:23,235 --> 00:01:26,405 AND HOW THEY MAY ACTIVATE 25 00:01:26,405 --> 00:01:31,710 REGENERATE SWITCH AND DR. 26 00:01:31,710 --> 00:01:34,947 MAZUNO'S PRESENT TATATION IS 27 00:01:34,947 --> 00:01:44,023 TIT 28 00:01:44,023 --> 00:01:44,256 TITLED. 29 00:01:44,256 --> 00:01:47,760 >> HELLO, EVERYONE FOR COMING 30 00:01:47,760 --> 00:01:52,031 HERE AND CHEERING ME UP HERE AND 31 00:01:52,031 --> 00:01:54,199 I AM APPOINTED WITH NIAMS AND 32 00:01:54,199 --> 00:01:58,470 TODAY I WANT TO INTRODUCE MY 33 00:01:58,470 --> 00:02:01,807 SCIENCE TO THE COMMUNITY. 34 00:02:01,807 --> 00:02:05,944 TITLE IS INTEGRATIVE STRUCTURE 35 00:02:05,944 --> 00:02:08,013 OF CELL BIOLOGY AND CHRIS 36 00:02:08,013 --> 00:02:10,215 MENTIONED I HAVE A LITTLE OF A 37 00:02:10,215 --> 00:02:13,118 SPECIAL OR UNIQUE BACKGROUND AND 38 00:02:13,118 --> 00:02:17,589 MY LAB WAS FOUNDED IN GERMANY IN 39 00:02:17,589 --> 00:02:21,360 MAX INSTITUTE OF BIOCHEMISTRY IN 40 00:02:21,360 --> 00:02:22,928 MUNICH. THIS IS ACTIVITY 41 00:02:22,928 --> 00:02:24,596 OCTOBER 1ST THAT WAS A RETREAT 42 00:02:24,596 --> 00:02:27,499 AND QUITE A FUN TIME OVER THERE. 43 00:02:27,499 --> 00:02:29,668 BUT I ACTUALLY DID POST GRAD NIH 44 00:02:29,668 --> 00:02:32,037 AND REALLY LIKED THE CULTURE AND 45 00:02:32,037 --> 00:02:35,841 HOW THINGS WORK HERE AND REALLY 46 00:02:35,841 --> 00:02:38,210 DREAMED OF COMING BACK TO NIH. 47 00:02:38,210 --> 00:02:40,279 WE MADE A DECISION TO MOVE TO 48 00:02:40,279 --> 00:02:45,718 NIH IN 2019 AND MOVE WAS TO 2020 49 00:02:45,718 --> 00:02:46,919 WHICH WAS A LITTLE DIFFICULT. 50 00:02:46,919 --> 00:02:50,522 I WILL TELL YOU ABOUT MY 51 00:02:50,522 --> 00:02:52,558 PANDEMIC MOVE LATER. 52 00:02:52,558 --> 00:02:55,728 THIS IS THE LAB IN 2024 AND 53 00:02:55,728 --> 00:02:58,230 RECENT PICTURE AND EVERYTHING IS 54 00:02:58,230 --> 00:02:59,631 SMOOTH AND NICE AND THANKS TO 55 00:02:59,631 --> 00:03:04,336 EVERYBODY WHO HELPED US TO 56 00:03:04,336 --> 00:03:14,880 ESTABLISH OR COMPLETE THE MOVE 57 00:03:15,114 --> 00:03:17,516 THE TECHNOLOGY IS INTEGRATED 58 00:03:17,516 --> 00:03:19,184 BIOLOGY AND QUESTION APPROACH 59 00:03:19,184 --> 00:03:20,619 LOOKING AT THINGS WITH 60 00:03:20,619 --> 00:03:22,988 TECHNOLOGY AND I MYSELF IS 61 00:03:22,988 --> 00:03:26,692 ORIGINALLY KRIO 62 00:03:26,692 --> 00:03:30,329 ELECTROMICROSCOPY SINCE 1999 AND 63 00:03:30,329 --> 00:03:32,030 BACHELORS MATHEMATIC STUDENT I 64 00:03:32,030 --> 00:03:33,866 DID IMAGE ANALYSIS AND YOU CAN 65 00:03:33,866 --> 00:03:38,337 SEE HERE THIS IS A TYPICAL KRIO 66 00:03:38,337 --> 00:03:41,373 EM STRUCTURE THAT YOU CAN GET. 67 00:03:41,373 --> 00:03:42,775 SO, WE STILL WELCOME THIS KIND 68 00:03:42,775 --> 00:03:45,744 OF A STRUCTURE AND IT IS A SUPER 69 00:03:45,744 --> 00:03:50,115 FINE STRUCTURE ANALYSIS AND OVER 70 00:03:50,115 --> 00:03:55,554 TIME MY AIM OF RUNNING A LAB IS 71 00:03:55,554 --> 00:03:57,623 TO [INDISCERNIBLE] PEOPLES 72 00:03:57,623 --> 00:03:59,625 CREATIVITY AND SETTING UP 73 00:03:59,625 --> 00:04:01,093 INFRASTRUCTURE SO PEOPLE CAN 74 00:04:01,093 --> 00:04:03,028 WORK WITH THEIR MIND AND 75 00:04:03,028 --> 00:04:03,362 CREATIVITY. 76 00:04:03,362 --> 00:04:07,800 SO YOU CAN SEE THIS IS A 77 00:04:07,800 --> 00:04:09,034 MOLECULAR ORGANIZATION 78 00:04:09,034 --> 00:04:11,436 OBSERVATION IN A CELLULAR AND 79 00:04:11,436 --> 00:04:13,906 CELL MORPHOLOGY AND MOVING TO 80 00:04:13,906 --> 00:04:16,041 TISSUE ORGANIZATION INVIFO THAT 81 00:04:16,041 --> 00:04:18,310 IS DIFFICULT FOR US AND WE HAVE 82 00:04:18,310 --> 00:04:19,478 COLLABORATION AND THIS IS 83 00:04:19,478 --> 00:04:23,148 BEAUTIFUL AND YOU SEE ELEGANCE 84 00:04:23,148 --> 00:04:25,851 IMAGE AND [INDISCERNIBLE] BY 85 00:04:25,851 --> 00:04:29,588 KEVIN'S LAB AT NIDDK A WONDERFUL 86 00:04:29,588 --> 00:04:32,057 COLLABORATION TO HAVE A REALLY 87 00:04:32,057 --> 00:04:33,091 COMPREHENSIVE UNDERSTANDING WHAT 88 00:04:33,091 --> 00:04:35,460 WE WANT TO SEE. 89 00:04:35,460 --> 00:04:37,296 OKAY. SO TALKING ABOUT IMAGING, 90 00:04:37,296 --> 00:04:40,599 I WANT TO TELL YOU A LITTLE 91 00:04:40,599 --> 00:04:41,600 ABOUT THE TECHNOLOGY DEVELOPMENT 92 00:04:41,600 --> 00:04:43,902 THAT IS HAPPENING IN THE IMAGING 93 00:04:43,902 --> 00:04:44,536 FIELD. 94 00:04:44,536 --> 00:04:47,906 YOU CAN SEE HERE THIS IS THE 95 00:04:47,906 --> 00:04:52,044 SIZE OR THE STRUCTURE RESOLUTION 96 00:04:52,044 --> 00:04:55,380 RANGE THAT YOU CAN ASK BY 97 00:04:55,380 --> 00:04:56,215 MICROSCOPIC TECHNIQUE AND THIS 98 00:04:56,215 --> 00:04:57,816 IS CONTEXT YOU CAN ASK. 99 00:04:57,816 --> 00:05:00,018 IF YOU WANT TO SEE A LARGE 100 00:05:00,018 --> 00:05:02,988 OBJECT, YOU WOULD GO WITH A 101 00:05:02,988 --> 00:05:05,791 CONVENTIONAL ELECTROMICROSCOPE 102 00:05:05,791 --> 00:05:08,227 OR LIGHT MICROSCOPE TO GET AN 103 00:05:08,227 --> 00:05:08,727 OVERVIEW. 104 00:05:08,727 --> 00:05:10,596 HOWEVER, YOU ARE MISSING 105 00:05:10,596 --> 00:05:12,030 MOLECULAR RESOLUTION YOU CAN SEE 106 00:05:12,030 --> 00:05:13,932 HERE IS MISSING AND TO 107 00:05:13,932 --> 00:05:16,034 COMPLEMENT THAT YOU WOULD GO 108 00:05:16,034 --> 00:05:20,072 WITH CRYSTALO GRAPHY OR NMR TO 109 00:05:20,072 --> 00:05:22,007 GIVE ATOMIC RESOLUTION 110 00:05:22,007 --> 00:05:22,441 INFORMATION. 111 00:05:22,441 --> 00:05:23,976 HOWEVER, YOU CAN SEE HERE 112 00:05:23,976 --> 00:05:29,147 IN-BETWEEN THIS IS MISSING UNTIL 113 00:05:29,147 --> 00:05:31,817 2014 WHEN TECHNOLOGICAL 114 00:05:31,817 --> 00:05:33,552 REVOLUTION WE CALL RESOLUTION 115 00:05:33,552 --> 00:05:34,620 HAPPENING IN E M FIELD. 116 00:05:34,620 --> 00:05:39,424 YOU CAN SEE HERE, NOW THE SINGLE 117 00:05:39,424 --> 00:05:40,926 PARTICLE CRYO-EM AND MOLECULE 118 00:05:40,926 --> 00:05:43,462 AND IMAGE ANALYSIS AND GETTING 119 00:05:43,462 --> 00:05:45,264 NEAR ATOMIC RESOLUTION 120 00:05:45,264 --> 00:05:48,033 INFORMATION OR RELATIVELY LARGE 121 00:05:48,033 --> 00:05:52,037 OR LARGER CONTEXT MOLECULE SUCH 122 00:05:52,037 --> 00:05:55,140 AS MICROMOLECULES SUCH AS 123 00:05:55,140 --> 00:05:56,275 COMPLEXES AND FURTHERMORE, 124 00:05:56,275 --> 00:05:59,044 CURRENT REVOLUTION IS HAPPENING 125 00:05:59,044 --> 00:06:03,048 WITH THE INSITU KRIO 126 00:06:03,048 --> 00:06:05,784 ELECTRO-TOMO GRAPHY AND GETTING 127 00:06:05,784 --> 00:06:06,985 CONTEXTUAL VIEW LOOKING AT CELL 128 00:06:06,985 --> 00:06:12,024 OR MOLECULES IN THE CELL TO 129 00:06:12,024 --> 00:06:13,692 HOPEFULLY NEAR OUGHTONOMIC 130 00:06:13,692 --> 00:06:15,827 RESOLUTION THAT IS ONGOING AND 131 00:06:15,827 --> 00:06:16,862 CONCEPTION THING THAT WE ARE 132 00:06:16,862 --> 00:06:19,965 DOING AND WE ARE GETTING SLOWLY 133 00:06:19,965 --> 00:06:22,801 BUT SURELY MOVING TOWARDS THAT 134 00:06:22,801 --> 00:06:23,368 DIRECTION. 135 00:06:23,368 --> 00:06:26,505 AND TALKING ABOUT INTEGRATIVE 136 00:06:26,505 --> 00:06:28,607 STRUCTURE BIOLOGY, WE WANT TO 137 00:06:28,607 --> 00:06:30,709 GIVE ANOTHER DIMENSION AND NOT 138 00:06:30,709 --> 00:06:33,378 ONLY THIS SPECTRUM BUT WE WILL 139 00:06:33,378 --> 00:06:35,380 ADD MORE FACTORS HERE. 140 00:06:35,380 --> 00:06:38,116 FOR EXAMPLE, WE CAN TALK ABOUT 141 00:06:38,116 --> 00:06:41,520 DYNAMICS AND INVIFO IMAGING TO 142 00:06:41,520 --> 00:06:43,855 BE COMBINED WITH THESE STATIC 143 00:06:43,855 --> 00:06:46,558 TECHNOLOGIES OR DISEASE CONTEXT 144 00:06:46,558 --> 00:06:48,060 WE CAN INCORPORATE OR FURTHER 145 00:06:48,060 --> 00:06:52,030 MORE HOW WE CAN LEARN OR APPLY 146 00:06:52,030 --> 00:06:53,532 THE CONTEXT OF WHAT WE HAVE DONE 147 00:06:53,532 --> 00:06:56,201 WITH THE APPLICATION. FOR 148 00:06:56,201 --> 00:07:00,038 EXAMPLE, TRANSLATIONAL ASPECT OF 149 00:07:00,038 --> 00:07:01,306 IT. 150 00:07:01,306 --> 00:07:03,875 SO, WITH THAT SAID I WANT TO 151 00:07:03,875 --> 00:07:06,845 TELL WHAT WE ARE DOING AND BIG 152 00:07:06,845 --> 00:07:09,781 APPROACH QUESTION WE TRY TO 153 00:07:09,781 --> 00:07:12,050 UNDERSTAND IT IS A DECISION THAT 154 00:07:12,050 --> 00:07:14,386 IS A FORMATION THAT IS CONNECTED 155 00:07:14,386 --> 00:07:16,021 TO DECISION MAKING PROCESS. 156 00:07:16,021 --> 00:07:17,889 YOU CAN SEE HERE FOR EXAMPLE 157 00:07:17,889 --> 00:07:20,859 THIS IS A PLATELET THAT IS 158 00:07:20,859 --> 00:07:23,028 ACTIVATED IN THE CONTEXT AND THE 159 00:07:23,028 --> 00:07:26,298 WOUND IS DETECTED AND ACTIVATION 160 00:07:26,298 --> 00:07:27,733 WOULD HAPPEN AND CELL SHAPE 161 00:07:27,733 --> 00:07:29,368 CHANGES OR SEE DIVIDING CELL 162 00:07:29,368 --> 00:07:31,770 THAT IS COORDINATING AND SO THAT 163 00:07:31,770 --> 00:07:34,139 THE COMPONENTS ARE DIVIDED 164 00:07:34,139 --> 00:07:36,675 EQUALLY AND IT GETS SHAPE 165 00:07:36,675 --> 00:07:39,344 CHANGES AND DIVISION HAPPENS. 166 00:07:39,344 --> 00:07:41,813 MOST PROMINENT EXAMPLE YOU CAN 167 00:07:41,813 --> 00:07:46,118 SEE WITH CELL SHAPE CHANGE IS 168 00:07:46,118 --> 00:07:48,520 NEURONAL CELL AND NEURON IS MOST 169 00:07:48,520 --> 00:07:50,889 POLARIZED CELL WE KNOW TO HAVE A 170 00:07:50,889 --> 00:07:54,326 METER LONG AXON AND FROM ONE 171 00:07:54,326 --> 00:07:56,862 SIDE TO THE OTHER SIDE OF THE 172 00:07:56,862 --> 00:07:58,163 NEURON FUNCTION IS COMPLETELY 173 00:07:58,163 --> 00:08:02,000 DIFFERENT AND EXAMPLES WE WORK 174 00:08:02,000 --> 00:08:08,040 ON AND NOT ONLY THAT IN HOMEO 175 00:08:08,040 --> 00:08:11,610 STASUS INFORMATION IS DISTURBED 176 00:08:11,610 --> 00:08:16,048 AND PROBLEM CANCER CELL 177 00:08:16,048 --> 00:08:19,217 FORMATION OR SICKLE CELL 178 00:08:19,217 --> 00:08:20,852 FORMATION AND APPLICATION YOU 179 00:08:20,852 --> 00:08:24,322 CAN TRY TO MAKE REGENERATE. 180 00:08:24,322 --> 00:08:26,391 THESE ARE THINGS NEW CHALLENGE 181 00:08:26,391 --> 00:08:28,794 FOR OUR LAB WE CAME FROM BASIC 182 00:08:28,794 --> 00:08:31,163 SCIENCE LAB AND BASIC SCIENCE 183 00:08:31,163 --> 00:08:32,898 INSTITUTE IN GERMANY AND MOVED 184 00:08:32,898 --> 00:08:37,369 TO NIH AND IS OPPORTUNITY TO 185 00:08:37,369 --> 00:08:41,106 ALSO ADDRESS HUMAN HEALTH 186 00:08:41,106 --> 00:08:41,940 PROBLEM. 187 00:08:41,940 --> 00:08:44,476 OKAY. SO NOW HOW WE DO IT, YOU 188 00:08:44,476 --> 00:08:46,478 CAN SEE FROM THE LEFT SIDE TO 189 00:08:46,478 --> 00:08:48,880 THE RIGHT SIDE THAT WE CALL TOP 190 00:08:48,880 --> 00:08:51,016 DOWN, WHICH MEANS WE START TO 191 00:08:51,016 --> 00:08:53,585 LOOK AT LARGE CONTEXT AND TRY TO 192 00:08:53,585 --> 00:08:56,154 REALLY GO USING A MICROSCOPE 193 00:08:56,154 --> 00:08:59,624 INTO THE MOLECULAR VIEW AND 194 00:08:59,624 --> 00:09:02,427 CRYOELECTRON TOMMO GRAPHY 195 00:09:02,427 --> 00:09:05,097 RECONSTRUCTION OF THE POLL HERE 196 00:09:05,097 --> 00:09:08,433 AND ALSO OUT GROSS CON OF THE 197 00:09:08,433 --> 00:09:10,635 NEURON AND TO, FOR EXAMPLE, 198 00:09:10,635 --> 00:09:14,272 SHOWING EXAMPLE OF PLAYERS DOING 199 00:09:14,272 --> 00:09:16,842 THE [INDISCERNIBLE] INFORMATION 200 00:09:16,842 --> 00:09:22,848 AT TIP OF THE GROWTH 201 00:09:22,848 --> 00:09:24,316 [INDISCERNIBLE] OR BOTTOM UP 202 00:09:24,316 --> 00:09:27,052 INSTITUTION MEANS WE CAN COLLECT 203 00:09:27,052 --> 00:09:28,453 PARTICLES OF INTEREST AND TRY TO 204 00:09:28,453 --> 00:09:29,521 UNDERSTAND EACH REGULATION OF 205 00:09:29,521 --> 00:09:31,990 EACH OF THE COMPLEXES OF 206 00:09:31,990 --> 00:09:33,425 MOLECULES AND TRY TO 207 00:09:33,425 --> 00:09:35,460 RECONSTITUTE INTO THE FUNCTIONAL 208 00:09:35,460 --> 00:09:38,997 UNITS SO THINGS START TO MAKE 209 00:09:38,997 --> 00:09:40,031 SENSE FUNCTIONALLY. 210 00:09:40,031 --> 00:09:44,236 SO, NOW, THIS IS AN EXAMPLE OF 211 00:09:44,236 --> 00:09:46,905 BOTTOM UP CONSTITUTION. I WON'T 212 00:09:46,905 --> 00:09:48,673 TALK ABOUT BOTTOM UP 213 00:09:48,673 --> 00:09:50,175 CONSTITUTION TOO MUCH HERE 214 00:09:50,175 --> 00:09:50,442 TODAY. 215 00:09:50,442 --> 00:09:52,043 YOU SEE THESE ARE PROTEINS OR 216 00:09:52,043 --> 00:09:53,778 MOLECULES WE HAVE WORKED ON FOR 217 00:09:53,778 --> 00:09:57,415 THE LAST 10 YEARS AND YOU CAN 218 00:09:57,415 --> 00:09:59,918 SEE THESE HAVE ITS OWN 219 00:09:59,918 --> 00:10:01,853 INTERESTING THINGS GOING ON AND 220 00:10:01,853 --> 00:10:03,755 WHEN WE KNOW WHAT REGULATION OF 221 00:10:03,755 --> 00:10:05,390 PROTEINS ARE WE CAN COMBINE THEM 222 00:10:05,390 --> 00:10:05,790 ALSO. 223 00:10:05,790 --> 00:10:09,461 THIS IS AN EXAMPLE OF THIS IS 224 00:10:09,461 --> 00:10:10,695 INTEGRATING AN IMPORTANT 225 00:10:10,695 --> 00:10:12,030 RECEPTOR PROTEIN DECIDING THE 226 00:10:12,030 --> 00:10:14,533 DECISION OF THE CELL. 227 00:10:14,533 --> 00:10:18,370 YOU SEE THE PLASMA MEMBRANE AND 228 00:10:18,370 --> 00:10:20,438 EXTRA CELLULAR LYINGON THAT IS 229 00:10:20,438 --> 00:10:22,641 ACTIVATING IT FROM INSIDE OF THE 230 00:10:22,641 --> 00:10:24,342 CELL YOU SEE THIS IS TAILING 231 00:10:24,342 --> 00:10:28,246 WHICH IS INTRACELLULAR ACTIVATOR 232 00:10:28,246 --> 00:10:30,015 OF [INDISCERNIBLE] THAT IS 233 00:10:30,015 --> 00:10:31,349 POKING FROM INSIDE OF THE CELL 234 00:10:31,349 --> 00:10:35,053 AND IS ATTACHED BY BOUNDING OF 235 00:10:35,053 --> 00:10:37,322 ACTIN AND MAKING THIS FORMATION 236 00:10:37,322 --> 00:10:40,158 AND WE TRY TO RECONSTITUTE SUCH 237 00:10:40,158 --> 00:10:42,928 STRUCTURE USING IN VITRO BOTTOM 238 00:10:42,928 --> 00:10:44,496 UP RECONSTITUTION. 239 00:10:44,496 --> 00:10:47,132 YOU SEE HERE WE CAN LOCK THE 240 00:10:47,132 --> 00:10:49,100 INTERMEDIATE PROCESS AND TO 241 00:10:49,100 --> 00:10:50,702 REALLY UNDERSTAND MOLECULAR 242 00:10:50,702 --> 00:10:52,037 ORGANIZATION AND MECHANISM OF 243 00:10:52,037 --> 00:10:53,772 HOW ACTIVATION WOULD BE 244 00:10:53,772 --> 00:10:54,239 HAPPENING. 245 00:10:54,239 --> 00:10:57,576 SO THIS IS A STRENGTH OF IN 246 00:10:57,576 --> 00:10:58,176 VITRO RECONSTITUTION WE HAVE 247 00:10:58,176 --> 00:11:01,346 FULL CONTROL OF THE MOLECULE OF 248 00:11:01,346 --> 00:11:01,746 INTEREST. 249 00:11:01,746 --> 00:11:03,248 NOW, NOT ONLY IS THE STRUCTURE 250 00:11:03,248 --> 00:11:05,150 PART, WE CAN ALSO LOOK INTO 251 00:11:05,150 --> 00:11:06,851 DYNAMIC ASPECT OF IT. 252 00:11:06,851 --> 00:11:09,454 THIS EXAMPLE I WANT PT TO SHOW 253 00:11:09,454 --> 00:11:13,758 YOU IS THESE PROTEINS 254 00:11:13,758 --> 00:11:15,460 ENCAPSULATED INSIDE OF THIS 255 00:11:15,460 --> 00:11:17,729 VESICLE THAT IS MIMICKING 256 00:11:17,729 --> 00:11:18,863 INTERMEMBRANE SURFACE OF THE 257 00:11:18,863 --> 00:11:20,632 CELL AND SEE BASICALLY THE GOAL 258 00:11:20,632 --> 00:11:23,335 OF THIS WAS TO ACTIVATE THE 259 00:11:23,335 --> 00:11:25,103 COMPLEX ENOUGH OF ACTIN SO THIS 260 00:11:25,103 --> 00:11:29,441 EXPERIMENT COULD BE PERFORMED ON 261 00:11:29,441 --> 00:11:31,509 PERIPHERY OF THIS. 262 00:11:31,509 --> 00:11:34,346 THIS IS A COMPLETE VIEW OF 263 00:11:34,346 --> 00:11:37,849 CONFIRMATION OF IN VITRO 264 00:11:37,849 --> 00:11:38,216 RECONSTITUTION. 265 00:11:38,216 --> 00:11:41,319 NOW, I'M GOING TO FOCUS MY STORY 266 00:11:41,319 --> 00:11:43,655 TO THE TOP DOWN AND INTRODUCE 267 00:11:43,655 --> 00:11:44,656 MORE DEEP ABOUT THE RESEARCH 268 00:11:44,656 --> 00:11:48,326 THAT IS GOING ON. 269 00:11:48,326 --> 00:11:50,929 SO, BEFORE TELLING YOU ABOUT THE 270 00:11:50,929 --> 00:11:52,797 TOP-DOWN APPROACH I WANT TO 271 00:11:52,797 --> 00:11:56,534 INTRODUCE ABOUT CRYOELECTRO-TOMO 272 00:11:56,534 --> 00:11:58,103 GRAPHY A NEWLY EMERGING 273 00:11:58,103 --> 00:11:59,404 TECHNIQUE I TOLD YOU ABOUT 274 00:11:59,404 --> 00:12:00,739 ALREADY. THIS IS A VIDEO CLIP 275 00:12:00,739 --> 00:12:03,041 THAT INTRODUCE THE TECHNOLOGY 276 00:12:03,041 --> 00:12:06,344 AND THIS IS YOUR CELL OF 277 00:12:06,344 --> 00:12:09,214 INTEREST AND MOLECULE ANYTHING 278 00:12:09,214 --> 00:12:12,017 THAT COULD BE IMBEDDED TO EM 279 00:12:12,017 --> 00:12:23,295 GRADE. SORRY I WILL TAGAIN. 280 00:12:39,010 --> 00:12:40,178 THIS TINY LITTLE THING CAN GO 281 00:12:40,178 --> 00:12:43,415 INSIDE OF A HUGE MICROSCOPE AND 282 00:12:43,415 --> 00:12:45,150 MAKING A [INDISCERNIBLE] SERIES. 283 00:12:45,150 --> 00:12:47,852 IT IS ACUTING INSIDE OF THE 284 00:12:47,852 --> 00:12:51,589 MICROSCOPE AND THIS IMAGE CAN BE 285 00:12:51,589 --> 00:12:54,926 COMBINED IN TWO 3-D BY BACK 286 00:12:54,926 --> 00:12:58,663 PROJECTION AND YOU SEE WE GET 287 00:12:58,663 --> 00:13:00,031 3-D RECONSTRUCTION AND ADVANTAGE 288 00:13:00,031 --> 00:13:03,968 OF CRYO-TOMO GRAPHY IS YOU LOOK 289 00:13:03,968 --> 00:13:04,936 INTO DENSITY DIFFERENCE BETWEEN 290 00:13:04,936 --> 00:13:06,705 PROTEIN AND LIPID TO WATER 291 00:13:06,705 --> 00:13:10,241 DENSITY SO YOU REALLY GET 292 00:13:10,241 --> 00:13:20,552 EVERYTHING THERE. 293 00:13:26,725 --> 00:13:27,258 YOU COULD SEE THIS PRETTY 294 00:13:27,258 --> 00:13:28,927 ENVELOPE BUT NOTHING INSIDE. 295 00:13:28,927 --> 00:13:30,662 OKAY. SO TOP-DOWN 296 00:13:30,662 --> 00:13:31,896 VISUALIZATION, I WANT TO TELL 297 00:13:31,896 --> 00:13:34,466 YOU TWO STORIES THAT ARE GOING 298 00:13:34,466 --> 00:13:45,210 ON IN IN RECENT YEARS AND IN 299 00:13:49,080 --> 00:13:54,219 PRESENCE OF SARS CO-V2 AND THIS 300 00:13:54,219 --> 00:13:56,020 IS IN OUR LAB AND SO FAR A LOT 301 00:13:56,020 --> 00:13:58,623 AND I WILL TELL YOU SOMETHING 302 00:13:58,623 --> 00:13:59,858 ABOUT IT. 303 00:13:59,858 --> 00:14:02,894 SECTION IS NEWEST STORY ABOUT 304 00:14:02,894 --> 00:14:05,897 [INDISCERNIBLE] RESPONSE AND 305 00:14:05,897 --> 00:14:16,141 REGENERATION. 306 00:14:18,510 --> 00:14:19,911 WHAT HAPPENED IS WE WENT TO 307 00:14:19,911 --> 00:14:22,313 GERMANY 2020 AND FEBRUARY WE 308 00:14:22,313 --> 00:14:25,016 PACKED EVERYTHING AND WAS ABOUT 309 00:14:25,016 --> 00:14:26,251 TO SHIP IT. 310 00:14:26,251 --> 00:14:28,453 IT COULDN'T BE SHIPPED BECAUSE 311 00:14:28,453 --> 00:14:32,390 OF THE LOCK DOWN AND 7 MONTHS WE 312 00:14:32,390 --> 00:14:37,395 DIDN'T HAVE ANYTHING AND 313 00:14:37,395 --> 00:14:39,898 AFTERWARDS WE HAD EQUIPMENT IN 314 00:14:39,898 --> 00:14:41,566 THE BUILDING AND CONTRACT TO 315 00:14:41,566 --> 00:14:42,801 MOVE SO AND DIFFICULT TIME FOR 316 00:14:42,801 --> 00:14:47,672 US AND FINALLY TRIED TO FETCH 317 00:14:47,672 --> 00:14:50,241 STUDENTS FROM GERMANY TO UP AND 318 00:14:50,241 --> 00:14:53,912 WAS IMPOSSIBLE FOR ARRANGING THE 319 00:14:53,912 --> 00:14:57,348 VISA AND EXCEPT INVESTIGATOR 320 00:14:57,348 --> 00:14:59,050 GAVE VISA TO COVID RESEARCH AND 321 00:14:59,050 --> 00:15:01,920 EVERYTHING WAS SAID FOR US TO 322 00:15:01,920 --> 00:15:04,656 WORK ON COVID RESEARCH AND THIS 323 00:15:04,656 --> 00:15:08,793 IS STUDENT THAT WORKED ON 324 00:15:08,793 --> 00:15:09,894 PROJECT. 325 00:15:09,894 --> 00:15:11,062 OKAY? 326 00:15:11,062 --> 00:15:12,630 NOW, JUST AS A SUMMARY WHAT WE 327 00:15:12,630 --> 00:15:15,633 DIDN'T HAVE IN THE LAB WHEN WE 328 00:15:15,633 --> 00:15:16,935 STARTED ALMOST EVERYTHING AND 329 00:15:16,935 --> 00:15:19,070 BITS AND PIECES OF EVERYTHING 330 00:15:19,070 --> 00:15:22,307 AND PLASMA WAS PACKED AND LOST 331 00:15:22,307 --> 00:15:23,508 PROTEINS WERE LOST AND 332 00:15:23,508 --> 00:15:25,376 EVERYTHING WAS LOST AND WE HAD 333 00:15:25,376 --> 00:15:27,512 PERMISSION TO WORK ON COVID 334 00:15:27,512 --> 00:15:30,815 PROJECT AND THANKS TO DONOR AND 335 00:15:30,815 --> 00:15:37,856 TRIED WE COULD USE AND SPIPROTE 336 00:15:37,856 --> 00:15:40,592 WE COULD BUY AND INSTRUMENT WE 337 00:15:40,592 --> 00:15:42,927 COULD USE INCLUDING CRYO-EM AND 338 00:15:42,927 --> 00:15:44,362 LOOKING WHAT WE COULD CONTRIBUTE 339 00:15:44,362 --> 00:15:46,531 TO THE HEALTH CRISIS, YOU SEE 340 00:15:46,531 --> 00:15:48,833 HERE I DON'T THINK I NEED TO 341 00:15:48,833 --> 00:15:50,502 EXPLAIN MUCH ABOUT COVID AND WHY 342 00:15:50,502 --> 00:15:53,471 IT IS SO DEVASTATING. 343 00:15:53,471 --> 00:15:55,974 ONE THING WE KNOW IS ACTIVATION 344 00:15:55,974 --> 00:15:58,910 OF THE PLATELET COULD BE A BIG 345 00:15:58,910 --> 00:16:01,446 PROBLEM AND COVID WHEN IS VERY 346 00:16:01,446 --> 00:16:03,281 SEVERE YOU GET CLOTTING OF THE 347 00:16:03,281 --> 00:16:05,250 BLOOD AND WE THOUGHT WE COULD 348 00:16:05,250 --> 00:16:09,387 WORK ON THE PLATELET ON THE 349 00:16:09,387 --> 00:16:09,988 SPECIAL BACKGROUND OF THE 350 00:16:09,988 --> 00:16:11,389 [INDISCERNIBLE] AND KNOW WHAT IS 351 00:16:11,389 --> 00:16:12,357 HAPPENING WITH THE RECEPTOR AND 352 00:16:12,357 --> 00:16:14,425 WHAT IS HAPPENING WITH THE 353 00:16:14,425 --> 00:16:18,897 ACTIVATION PRO S PROCESS AND 354 00:16:18,897 --> 00:16:20,798 DECIDED TO WORK ON THAT AND 355 00:16:20,798 --> 00:16:21,866 SPIKE PROTEIN STRUCTURE AND 356 00:16:21,866 --> 00:16:24,335 NOTICE IT QUICKLY AND COULD GET 357 00:16:24,335 --> 00:16:25,803 NICE STRUCTURE AND GOOD QUALITY 358 00:16:25,803 --> 00:16:27,338 CONTROL FOR US. 359 00:16:27,338 --> 00:16:29,908 THEN WE LEARNED HOW TO PURIFY OR 360 00:16:29,908 --> 00:16:32,010 EXTRACT THE PLATELET OUT OF THE 361 00:16:32,010 --> 00:16:33,678 BLOOD AND MIX IT TOGETHER AND 362 00:16:33,678 --> 00:16:36,681 THAT IS AN S PROTEIN ON THE 363 00:16:36,681 --> 00:16:37,048 PLATELET. 364 00:16:37,048 --> 00:16:40,018 NOW YOU CAN SEE HERE THIS IS A 365 00:16:40,018 --> 00:16:41,986 LIGHT MICROSCOPIC ANALYSIS OF 366 00:16:41,986 --> 00:16:44,289 THE PLATELET OF THE CONTROL AND 367 00:16:44,289 --> 00:16:47,926 YOU SEE MORE OF THESE LEADERSHIP 368 00:16:47,926 --> 00:16:50,428 ARE NOT MUCH CHANGING AND 369 00:16:50,428 --> 00:16:52,030 HAPPILY AROUND AND SEE ACTIVATED 370 00:16:52,030 --> 00:16:53,831 ON THE SURFACE AND IN THE 371 00:16:53,831 --> 00:16:56,868 PRESENCE OF THE COVID SPIKE 372 00:16:56,868 --> 00:16:58,903 PROTEIN YOU CAN SEE LOTS OF THE 373 00:16:58,903 --> 00:17:00,872 ACTIVATION ACTUALLY ARE GOING 374 00:17:00,872 --> 00:17:01,239 ON. 375 00:17:01,239 --> 00:17:03,641 SO THIS YOU CAN SEE VISUALLY 376 00:17:03,641 --> 00:17:05,176 ALREADY THAT ACTIVATION WAS 377 00:17:05,176 --> 00:17:06,945 HAPPENING IN THE PRESENCE OF THE 378 00:17:06,945 --> 00:17:08,346 S PROTEIN AND MORE 379 00:17:08,346 --> 00:17:10,081 INTERESTINGLY, WE COULD SEE A 380 00:17:10,081 --> 00:17:12,750 LOT OF SPIKE -- A LOT OF 381 00:17:12,750 --> 00:17:14,252 PLATELETS THAT GET ATTACHED TO 382 00:17:14,252 --> 00:17:20,058 ONE SIDE OF THE ECM AND START TO 383 00:17:20,058 --> 00:17:21,092 PROVIDE MICROACTIVATED AND START 384 00:17:21,092 --> 00:17:22,760 TO GET STRETCHED THAT YOU CAN 385 00:17:22,760 --> 00:17:24,028 SEE HERE. 386 00:17:24,028 --> 00:17:25,897 THIS WAS QUITE INTERESTING 387 00:17:25,897 --> 00:17:27,298 MORPHOLOGY THAT WE SAW. 388 00:17:27,298 --> 00:17:29,033 WE QUANTIFIED IT AS YOU CAN SEE 389 00:17:29,033 --> 00:17:31,069 IN THE PRESENCE OF THE SPIKE WE 390 00:17:31,069 --> 00:17:34,505 REALLY SEE A LOT MORE OF THIS 391 00:17:34,505 --> 00:17:36,040 STRETCHED MORPHOLOGY. 392 00:17:36,040 --> 00:17:39,711 THIS WAS ALSO DOSE-DEPENDENT 393 00:17:39,711 --> 00:17:44,015 MEANING THERE IS DIRECT 394 00:17:44,015 --> 00:17:46,718 CORRELATION BETWEEN PRESENCE OF 395 00:17:46,718 --> 00:17:49,954 SARS CO-INDIVIDUAL FORMATION ON 396 00:17:49,954 --> 00:17:51,556 THE PLATELET. 397 00:17:51,556 --> 00:17:52,890 WE DECIDED TO HAVE A LOOK AT 398 00:17:52,890 --> 00:17:56,027 WHAT IS GOING ON THERE USINGCIO 399 00:17:56,027 --> 00:17:58,496 EM AND HERE WE PREPARE A 400 00:17:58,496 --> 00:17:59,964 PLATELET MIXING WITH SPIKE 401 00:17:59,964 --> 00:18:04,035 PROTEIN TO MAKE CRYO-EM GREAT 402 00:18:04,035 --> 00:18:05,670 [INDISCERNIBLE] AND HAVE A LOOK 403 00:18:05,670 --> 00:18:10,742 AT TOMMO GRAPHY AND YOU SEE HERE 404 00:18:10,742 --> 00:18:12,710 MEDIUM SNAPSHOT TO VISUALIZE 405 00:18:12,710 --> 00:18:13,778 OVERVIEW OF THE PLATELET AND YOU 406 00:18:13,778 --> 00:18:17,448 CAN SEE HERE THIS IS A COLLAGEN 407 00:18:17,448 --> 00:18:22,887 FIBER AND THIS IS ACTUALLY THE 408 00:18:22,887 --> 00:18:31,195 DISTORTED PLATELET THIS IS A 409 00:18:31,195 --> 00:18:31,863 CONTROL AND SEE THE EFFECT OF IT 410 00:18:31,863 --> 00:18:33,031 AND THERE IS DIRECT 411 00:18:33,031 --> 00:18:34,065 VISUALIZATION WE CAN DO. 412 00:18:34,065 --> 00:18:37,301 HERE YOU CAN SEE THIS IS THE 413 00:18:37,301 --> 00:18:40,605 AREA WE COULD COLLECT 414 00:18:40,605 --> 00:18:40,905 TOMOGRAPHY. 415 00:18:40,905 --> 00:18:44,842 SO THIS IS ONE EXAMPLE OF 416 00:18:44,842 --> 00:18:47,445 HUNDREDS OF TOMMO GRAMS THAT WE 417 00:18:47,445 --> 00:18:50,515 TAKE AND YOU SEE HERE THIS IS 418 00:18:50,515 --> 00:18:53,985 THE PLATELET AND FEEDO PODIUM 419 00:18:53,985 --> 00:18:58,322 HAPPENING WITH ACTIVE STRESS 420 00:18:58,322 --> 00:18:59,791 FIBER. YOU SEE HERE THE DIRECT 421 00:18:59,791 --> 00:19:00,958 ATTACHMENT OF THE SPIKE PROTEIN 422 00:19:00,958 --> 00:19:03,928 ON THE SURFACE OF THE MEMBRANE. 423 00:19:03,928 --> 00:19:06,197 WE CAN REALLY SEE QUITE A BIT OF 424 00:19:06,197 --> 00:19:09,467 THE S PROTEIN ATTACHED ON THE 425 00:19:09,467 --> 00:19:15,106 MEMBRANE AND IT IS CAUSING THE 426 00:19:15,106 --> 00:19:15,440 DEFORMATION. 427 00:19:15,440 --> 00:19:17,542 NOW, YOU MIGHT WONDER IF THIS IS 428 00:19:17,542 --> 00:19:19,410 A [INDISCERNIBLE] AND SURFACE 429 00:19:19,410 --> 00:19:21,612 RECEPTORS ARE MOSTLY INTEGRATED 430 00:19:21,612 --> 00:19:23,948 ON THE PLATELET AND LOOKING AT 431 00:19:23,948 --> 00:19:26,617 CONTROL, YOU DON'T SEE ANY OR 432 00:19:26,617 --> 00:19:29,020 NOT MUCH OF THE PROTRUSION OUT 433 00:19:29,020 --> 00:19:30,788 OF THE MEMBRANE SURFACE AND MUST 434 00:19:30,788 --> 00:19:33,558 BE THE PROTEIN THAT IS ATTACHED 435 00:19:33,558 --> 00:19:35,259 FROM THE OUTSIDE. 436 00:19:35,259 --> 00:19:39,130 SO TO VALIDATE THAT, WHAT WE DO 437 00:19:39,130 --> 00:19:42,266 IS CALLED SUB-TOMO GRAM 438 00:19:42,266 --> 00:19:45,770 AVERAGING AND COLLECT DENSITY 439 00:19:45,770 --> 00:19:47,939 AND 1,284 PARTICLES AND DID 440 00:19:47,939 --> 00:19:48,973 [INDISCERNIBLE] OF IT. 441 00:19:48,973 --> 00:19:51,209 IF YOU DO SOMETHING SIMILAR, 442 00:19:51,209 --> 00:19:53,978 LARGER AMOUNT ON TO-DO SINGLE 443 00:19:53,978 --> 00:19:55,480 PARTICLE WAY YOU WOULD GET 444 00:19:55,480 --> 00:19:57,181 ATOMIC STRUCTURE THAT I HAVE 445 00:19:57,181 --> 00:19:58,683 SHOWN YOU ALREADY. 446 00:19:58,683 --> 00:20:01,753 WE HAVE TOMOGRAPHY AND 447 00:20:01,753 --> 00:20:03,287 RESOLUTION THAT IS A LITTLE 448 00:20:03,287 --> 00:20:04,989 COMPROMISED BUT GOOD ENOUGH TO 449 00:20:04,989 --> 00:20:06,958 IDENTIFY WHAT IT IS. 450 00:20:06,958 --> 00:20:08,693 SO AFTER RECONSTRUCTION, YOU CAN 451 00:20:08,693 --> 00:20:11,095 SEE HERE THIS IS THE DENSITY 452 00:20:11,095 --> 00:20:12,029 THAT WE GOT. 453 00:20:12,029 --> 00:20:17,034 SO THIS SHOWS THE VERY TYPICAL 454 00:20:17,034 --> 00:20:18,336 TIME ARRANGEMENT OF THE SPIKE 455 00:20:18,336 --> 00:20:21,506 PROTEIN AND HERE ALSO YOU CAN 456 00:20:21,506 --> 00:20:30,081 SEE THAT OUR TOPOGRAPHIC AFTER 457 00:20:30,081 --> 00:20:34,585 OF ATOMIC STRUCTURE -- THE 458 00:20:34,585 --> 00:20:36,788 STRUCTURE ON THE S PROTEIN AND 459 00:20:36,788 --> 00:20:38,656 DOING MORE OF ANALYSIS WE COULD 460 00:20:38,656 --> 00:20:41,192 ALSO REALIZE SOME OF THE 461 00:20:41,192 --> 00:20:42,794 POPULATION OF THE SPIKE PROTEIN 462 00:20:42,794 --> 00:20:45,096 HAVE OPEN CONFIRMATION AND ONE 463 00:20:45,096 --> 00:20:47,732 OF THE RBD DOMAINS RECREPTOR 464 00:20:47,732 --> 00:20:50,067 BINDING DOMAIN IS STICKING UP 465 00:20:50,067 --> 00:20:51,936 AND LIFTING UP AND ATTACHING TO 466 00:20:51,936 --> 00:20:57,141 THE SURFACE OF THE PLACELET. 467 00:20:57,141 --> 00:20:58,476 SO, HAVING THIS RECONSTRUCTION 468 00:20:58,476 --> 00:21:02,780 AND ANALYSIS DONE, WHAT WE CAN 469 00:21:02,780 --> 00:21:06,350 DO IS SOMETHING CALLED BACK 470 00:21:06,350 --> 00:21:07,685 PROTEIN WE CAN PUT COORDINATE 471 00:21:07,685 --> 00:21:10,955 BACK OF THE ANALYSIS. 472 00:21:10,955 --> 00:21:13,624 YOU SEE HERE THIS IS AN S 473 00:21:13,624 --> 00:21:15,726 PROTEIN DIRECTLY ATTACHING TO 474 00:21:15,726 --> 00:21:18,729 THE MEMBRANE AND EACH OF THEM WE 475 00:21:18,729 --> 00:21:20,698 CAN REALLY UNDERSTAND HOW THIS 476 00:21:20,698 --> 00:21:23,868 PROTEIN IS ATTACHING TO THE 477 00:21:23,868 --> 00:21:25,036 PLATELET SURFACE AND TO THE 478 00:21:25,036 --> 00:21:27,171 INFORMATION WE GOT FROM THAT, 479 00:21:27,171 --> 00:21:29,974 YOU CAN SEE THE CARVED PLATELET 480 00:21:29,974 --> 00:21:32,009 AND SPIKE PROTEIN ATTACHED TO 481 00:21:32,009 --> 00:21:33,377 IT. 482 00:21:33,377 --> 00:21:34,846 OBVIOUSLY, IT LOOKS LIKE WHEN 483 00:21:34,846 --> 00:21:37,048 CURVATURE OF THE MEMBRANE IS 484 00:21:37,048 --> 00:21:39,383 HIGHER THAN YOU SEE MORE OF THE 485 00:21:39,383 --> 00:21:42,386 SPIKE PROTEIN WHICH WE 486 00:21:42,386 --> 00:21:44,188 QUANTIFIED AND AVERAGE COULD BE 487 00:21:44,188 --> 00:21:46,057 QUANTIFIED AND YOU CAN ALSO 488 00:21:46,057 --> 00:21:47,491 CORRELATE THAT TO THE DENSITY OF 489 00:21:47,491 --> 00:21:49,961 THE SPIKE PROTEIN. 490 00:21:49,961 --> 00:21:52,396 THIS INDICATES THAT MORE OF THE 491 00:21:52,396 --> 00:21:55,233 CURVED PART IS PROTEIN TO ATTACH 492 00:21:55,233 --> 00:22:00,338 AND IT ALSO INDICATES THAT THE 493 00:22:00,338 --> 00:22:01,539 POED YA INFORMATION HIGH 494 00:22:01,539 --> 00:22:04,575 CURVATURE S BINDING TO THE 495 00:22:04,575 --> 00:22:06,043 PLATELET CAUSES SUPERCURVATURE 496 00:22:06,043 --> 00:22:12,016 HAVING ACTIVATION OF THE -- FOR 497 00:22:12,016 --> 00:22:13,517 THE PLATELET. NEXT INFORMATION 498 00:22:13,517 --> 00:22:17,088 THAT WE COULD GET AS YOU CAN SEE 499 00:22:17,088 --> 00:22:19,790 HERE, ORIENTATION OF THE SPIKE 500 00:22:19,790 --> 00:22:21,559 ON THE MEMBRANE SEEMS TO DIFFER 501 00:22:21,559 --> 00:22:23,794 FROM EACH OTHER AND COULD TAKE 502 00:22:23,794 --> 00:22:25,396 STATISTICS OF IT AND WE CAN SEE 503 00:22:25,396 --> 00:22:27,365 THAT A SPIKE PROTEIN IS NOT 504 00:22:27,365 --> 00:22:31,002 REALLY BINDING ON MEMBRANE IN A 505 00:22:31,002 --> 00:22:33,337 ROBUST WAY BUT WOBBLES AROUND 506 00:22:33,337 --> 00:22:36,307 AND NOT A SURPRISE AND THIS IS A 507 00:22:36,307 --> 00:22:37,675 TRIMER AND THERE ARE 508 00:22:37,675 --> 00:22:38,709 OPPORTUNITIES FOR THE SPIKE 509 00:22:38,709 --> 00:22:41,045 PROTEIN TO BE BOUND TO THE 510 00:22:41,045 --> 00:22:42,947 MEMBRANE SURFACE AND WE CAN 511 00:22:42,947 --> 00:22:44,682 VISUALIZE THAT BY DIRECTLY 512 00:22:44,682 --> 00:22:46,884 LOOKING AT SPIKE PROTEIN 513 00:22:46,884 --> 00:22:48,352 ATTACHING ON THE MEMBRANE 514 00:22:48,352 --> 00:22:49,954 SURFACE THAT WE THINK IS THE WAY 515 00:22:49,954 --> 00:22:52,089 OF MAXIMIZING THE SPIKE PROTEIN 516 00:22:52,089 --> 00:22:57,028 TO RECOGNIZE THE SURFACE OF THE 517 00:22:57,028 --> 00:22:57,295 PLATELET. 518 00:22:57,295 --> 00:23:01,198 SOMETHING SIMILAR WAS OBSERVED. 519 00:23:01,198 --> 00:23:03,034 BOTTOM PART OF THE SPIKE COMING 520 00:23:03,034 --> 00:23:05,703 TO THE CORE OF THE VIRUS. 521 00:23:05,703 --> 00:23:08,039 YOU SEE THEY ARE WOBBLING AROUND 522 00:23:08,039 --> 00:23:09,707 AND WE HAVE [INDISCERNIBLE] OF 523 00:23:09,707 --> 00:23:11,575 THE SPIKE PROTEIN THAT WOULD 524 00:23:11,575 --> 00:23:14,111 MAXIMIZE FOR THAT ATTACHMENT OF 525 00:23:14,111 --> 00:23:16,514 THE VIRUS TO THE HOST. 526 00:23:16,514 --> 00:23:19,016 SO NOW WHAT KIND OF A RECEPTOR 527 00:23:19,016 --> 00:23:20,651 IS THERE TO RECOGNIZE THE SPIKE 528 00:23:20,651 --> 00:23:23,187 PROTEIN AND WE HAVE A GOOD 529 00:23:23,187 --> 00:23:24,689 HUNCH. SO MANY OF THE PRO E- 530 00:23:24,689 --> 00:23:26,524 TEENS ARE BINDING ON THE 531 00:23:26,524 --> 00:23:28,025 MEMBRANE AND WHAT WOULD BE 532 00:23:28,025 --> 00:23:29,927 MOSTLY EXPRESSED AND THAT WOULD 533 00:23:29,927 --> 00:23:32,029 BE INTEGRATING. 534 00:23:32,029 --> 00:23:36,033 ALSO, INTERESTINGLY, YOU CAN SEE 535 00:23:36,033 --> 00:23:39,570 THIS RGD PEPTIDE OR SEQUENCE 536 00:23:39,570 --> 00:23:45,810 THAT IS INCORPORATED INTO SARS 537 00:23:45,810 --> 00:23:50,681 CO-V2 OR [INDISCERNIBLE] AS 538 00:23:50,681 --> 00:23:53,417 CORONAVIRUS DOESN'T HAVE LGD 539 00:23:53,417 --> 00:23:58,589 THAT IS A FAMOUS STRETCH OF THE 540 00:23:58,589 --> 00:24:00,024 ACID KNOWN AS [INDISCERNIBLE]. 541 00:24:00,024 --> 00:24:03,894 WE ALSO SYMPTOMS OF THE 542 00:24:03,894 --> 00:24:06,364 STRUCTURE OF THE INTEGRIN OPEN 543 00:24:06,364 --> 00:24:09,100 FORM OF PRESENCE OF FIBERMECTIN 544 00:24:09,100 --> 00:24:12,003 YOU SEE RGD THAT IS STICKING TO 545 00:24:12,003 --> 00:24:15,172 HEAD OF INTEGRIN THAT IS 546 00:24:15,172 --> 00:24:17,241 CONTRIBUTING TO ACTIVATION OF 547 00:24:17,241 --> 00:24:18,342 THE INTEGRIN AND SOMETHING 548 00:24:18,342 --> 00:24:20,311 SIMILAR MUST BE HAPPENING FOR 549 00:24:20,311 --> 00:24:22,613 INTEGRIN OF THE PLATELET AND USE 550 00:24:22,613 --> 00:24:25,216 THIS PURIFIED VERSION OF THE 551 00:24:25,216 --> 00:24:29,186 INTEGRIN ALPHA 3 BETA 3 TO BE 3 552 00:24:29,186 --> 00:24:34,992 BETA 3 AND ALPHA 5 BETA TESTING 553 00:24:34,992 --> 00:24:35,426 INTERACTION. 554 00:24:35,426 --> 00:24:38,496 YOU SEE HERE 5 BETA 1 DO HAVE 555 00:24:38,496 --> 00:24:39,930 AFFINITY TO THE SPIKE PROTEIN 556 00:24:39,930 --> 00:24:42,600 AND JUST AS A NOTE TO BE BETA 3 557 00:24:42,600 --> 00:24:45,903 IS THE MOST MAJOR INTEGRIN THAT 558 00:24:45,903 --> 00:24:49,473 IS EXPRESSED ON THE PLATELET. 559 00:24:49,473 --> 00:24:53,344 THAT ALSO EXPLAINS WHY THIS IS 560 00:24:53,344 --> 00:24:53,911 STOCHASTIC. 561 00:24:53,911 --> 00:24:58,582 IF THIS PROTEIN, S PROTEIN LOCKS 562 00:24:58,582 --> 00:25:01,218 ON TO BETA 3 IT WILL BE A BIG 563 00:25:01,218 --> 00:25:03,854 MUCH, MUCH BIGGER PROBLEM. 564 00:25:03,854 --> 00:25:05,089 ALL BLOOD GETS CLOTTED. 565 00:25:05,089 --> 00:25:08,259 THIS IS THE -- THE MECHANISM ON 566 00:25:08,259 --> 00:25:12,329 THAT WE DERIVED FROM THE 567 00:25:12,329 --> 00:25:14,732 OBSERVATION THAT SPIKE PROTEIN 568 00:25:14,732 --> 00:25:17,568 IS RECOGNIZING ON TO INTEGRIN 569 00:25:17,568 --> 00:25:23,207 SURFACE OF THE PLATELET IS 570 00:25:23,207 --> 00:25:28,879 STOCHASTIC INFORMATION -- IT 571 00:25:28,879 --> 00:25:32,016 COULD GO TO IRREVERSIBLE FORM OF 572 00:25:32,016 --> 00:25:34,785 THE PLATELET AND COULD GO BACK 573 00:25:34,785 --> 00:25:40,791 AND ONE PART IS ATTACHED TO ACM 574 00:25:40,791 --> 00:25:45,062 AND THIS IS A STUDY TO FINISH IT 575 00:25:45,062 --> 00:25:47,798 AND ARE HAPPY TO DO SOMETHING 576 00:25:47,798 --> 00:25:52,036 FOR AT LEAST THE PUBLIC HEALTH 577 00:25:52,036 --> 00:25:55,106 CRI 578 00:25:55,106 --> 00:25:55,573 CRISIS. 579 00:25:55,573 --> 00:26:00,511 WE WILL GO TO LONGSTANDING 580 00:26:00,511 --> 00:26:02,813 PROJECT ABOUT THE AXIUM OR THE 581 00:26:02,813 --> 00:26:05,382 NEURON STARTED SOME YEARS AGO 582 00:26:05,382 --> 00:26:09,553 ALREADY BACK IN GERMANY AND PHD 583 00:26:09,553 --> 00:26:13,424 STUDENT STARTED TO MAKE THE 584 00:26:13,424 --> 00:26:16,026 PIPELINE THAT WAS ALREADY SIX OR 585 00:26:16,026 --> 00:26:17,628 SEVEN YEARS AGO. 586 00:26:17,628 --> 00:26:19,997 WE WERE QUITE PIONEERING THE 587 00:26:19,997 --> 00:26:23,167 CURVATURE OF THE NEURON TO EM 588 00:26:23,167 --> 00:26:24,001 GRADE. 589 00:26:24,001 --> 00:26:27,271 YOU SEE HERE THIS IS MOST THAT 590 00:26:27,271 --> 00:26:32,710 WE GET EMBRYO AND THE PRIMARY 591 00:26:32,710 --> 00:26:36,013 CULTURE GOING TO EM GRADE AND WE 592 00:26:36,013 --> 00:26:39,617 GO TO CORRELATE AND CRYO-EM AND 593 00:26:39,617 --> 00:26:41,252 HAVE A LOOK AND GET A DEEPER 594 00:26:41,252 --> 00:26:41,619 VIEW. 595 00:26:41,619 --> 00:26:45,289 HAVING THIS DONE, THIS IS THE 596 00:26:45,289 --> 00:26:48,325 FIRST TIME THAT WE OBSERVED HOW 597 00:26:48,325 --> 00:26:52,029 ORGANIZATIONAL ORCHESTRATION OF 598 00:26:52,029 --> 00:26:56,033 THE MOLECULE AT AXIUM BRANCHING 599 00:26:56,033 --> 00:26:58,102 POINT AND THIS IS FIRST TAKE 600 00:26:58,102 --> 00:27:02,206 HOME STUDY AND MESSAGE WAS 601 00:27:02,206 --> 00:27:04,008 INTERESTING THAT [INDISCERNIBLE] 602 00:27:04,008 --> 00:27:07,578 WAS CONCENTRATED AT AXIUM BRANCH 603 00:27:07,578 --> 00:27:09,146 POINT AND WHERE IT GENERATED 604 00:27:09,146 --> 00:27:11,115 FROM AND LOTS OF PRODUCTIONS OF 605 00:27:11,115 --> 00:27:12,850 THE MATERIAL HAVE TO HAPPEN. 606 00:27:12,850 --> 00:27:15,653 THIS WAS OUR KIND OF SHOWCASE 607 00:27:15,653 --> 00:27:17,321 STUDY THAT IS PASSED ALREADY. 608 00:27:17,321 --> 00:27:19,690 I WON'T TALK ABOUT IT TODAY. 609 00:27:19,690 --> 00:27:23,627 TODAY, I WANT TO FOCUS ON THE 610 00:27:23,627 --> 00:27:27,598 STORY THAT IS RECENT ABOUT 611 00:27:27,598 --> 00:27:31,735 AXONAL GENERATION IN PRESENCE OF 612 00:27:31,735 --> 00:27:34,805 DRUG EPOBY AND PROJECT 613 00:27:34,805 --> 00:27:37,341 ORCHESTRATED AT NIH AND 614 00:27:37,341 --> 00:27:38,976 [INDISCERNIBLE] WHO IS A MEDICAL 615 00:27:38,976 --> 00:27:42,346 DOCTOR HAVING NO EXPERIENCE 616 00:27:42,346 --> 00:27:45,783 BEFORE COMING HERE AND HAD VERY 617 00:27:45,783 --> 00:27:48,118 GOOD VIEW AND GUTS AND FOCUS AND 618 00:27:48,118 --> 00:27:53,891 REALLY MANAGED TO HAVE THE 619 00:27:53,891 --> 00:27:55,192 STORYLINE GO FORWARD. 620 00:27:55,192 --> 00:27:59,363 AND ALSO IN VERY, VERY GREAT 621 00:27:59,363 --> 00:28:01,699 TEAM WHO FINISH UP THE STUDY AND 622 00:28:01,699 --> 00:28:03,767 KENNY HAD TO LEAVE TO GO BACK TO 623 00:28:03,767 --> 00:28:06,937 MEDICAL STUDY HE IS A NEUROCELL 624 00:28:06,937 --> 00:28:07,571 BIOLOGIST. 625 00:28:07,571 --> 00:28:10,874 JUST HAVE A LOOK AT THEIR 626 00:28:10,874 --> 00:28:12,009 EFFORT. 627 00:28:12,009 --> 00:28:13,911 SO, WIDER [INDISCERNIBLE] IN 628 00:28:13,911 --> 00:28:14,979 REGENERATION IS IMPORTANT. 629 00:28:14,979 --> 00:28:16,447 IT IS BECAUSE IN THE CONTEXT OF 630 00:28:16,447 --> 00:28:20,050 THE HEALTH, IT IS SOMETHING TO 631 00:28:20,050 --> 00:28:24,021 DO WITH THE BRAIN INJURES. 632 00:28:24,021 --> 00:28:32,563 TRAUMATIC BRAIN INJURY. 633 00:28:32,563 --> 00:28:35,833 WHAT IS HAPPENING IN YOUR BRAIN 634 00:28:35,833 --> 00:28:41,338 IS IN AXON. THIS IS A PROBLEM 635 00:28:41,338 --> 00:28:43,807 CENTRAL NEURAL SYSTEM BRAIN 636 00:28:43,807 --> 00:28:47,077 DOESN'T WANT TO REGENERATE AND 637 00:28:47,077 --> 00:28:48,779 DEEP PART OF THE BRAIN THAT WE 638 00:28:48,779 --> 00:28:51,448 WORK ON AND WHAT HAPPENS YOU GET 639 00:28:51,448 --> 00:28:53,150 UNCONSCIOUS AND HAVE COGNITIVE 640 00:28:53,150 --> 00:28:54,852 COGNITION PROBLEM OR EVENTUALLY 641 00:28:54,852 --> 00:28:55,419 YOU DIE. 642 00:28:55,419 --> 00:28:59,857 THIS IS SOMETHING THAT WE WANT 643 00:28:59,857 --> 00:29:02,960 TO ADDRESS AND ALSO POSSIBILITY 644 00:29:02,960 --> 00:29:05,696 OF THERAPEUTIC OPTIONS. 645 00:29:05,696 --> 00:29:07,998 MOLECULARLY OR CELL BIOLOGICALLY 646 00:29:07,998 --> 00:29:10,567 WE SEE INJURED AXON THAT IS 647 00:29:10,567 --> 00:29:10,868 FOLLOWING. 648 00:29:10,868 --> 00:29:13,937 YOU SEE HERE THAT THIS AXON IS 649 00:29:13,937 --> 00:29:20,044 LITERALLY CUT OFF AND MEANSNENTS 650 00:29:20,044 --> 00:29:22,079 COMPONENTS ARE LEAKED OUT AND 651 00:29:22,079 --> 00:29:23,480 [INDISCERNIBLE] WHICH IS 652 00:29:23,480 --> 00:29:27,251 DISRUPTED AND FIRST REACTION OF 653 00:29:27,251 --> 00:29:30,354 THE AXON IS TO CLOSE AND TO 654 00:29:30,354 --> 00:29:34,258 PROTECT SURROUNDING TISSUE AND 655 00:29:34,258 --> 00:29:35,826 CALCIUM GOES IN AND 656 00:29:35,826 --> 00:29:37,895 [INDISCERNIBLE] MICROTUBULE SO 657 00:29:37,895 --> 00:29:40,297 IT SHRINKS AND WAY THAT 658 00:29:40,297 --> 00:29:46,804 DEGENERATION OF THE NEURON IS 659 00:29:46,804 --> 00:29:47,104 HAPPENING. 660 00:29:47,104 --> 00:29:50,341 NOW, THIS IS A COLLABORATOR FROM 661 00:29:50,341 --> 00:29:53,510 DATA FROM 2015 AND STUDY WAS 662 00:29:53,510 --> 00:29:56,714 DONE IN PRESENCE OF DRUG EPOB 663 00:29:56,714 --> 00:30:00,351 WHICH IS INJECTED TO MOST THAT 664 00:30:00,351 --> 00:30:03,520 HAS SPINAL CORD GIRAFFE 8 WEEKS 665 00:30:03,520 --> 00:30:04,988 MOUSE CAN WORK NICELY AND SHOWS 666 00:30:04,988 --> 00:30:07,224 THIS DRUG IS WORKING. 667 00:30:07,224 --> 00:30:10,694 WHAT IS DRUG DOING? IT IS 668 00:30:10,694 --> 00:30:11,962 [INDISCERNIBLE] THAT IS A DRUG 669 00:30:11,962 --> 00:30:17,101 THAT WAS ORIGINALLY DEVELOPED AS 670 00:30:17,101 --> 00:30:19,002 ANTICANCER DRUG. 671 00:30:19,002 --> 00:30:23,040 IDEA IS THAT IT IS RECOGNIZING 672 00:30:23,040 --> 00:30:25,476 MICROTUBULE TO STABILIZE 673 00:30:25,476 --> 00:30:28,746 MICROTUBULE SO IT ARRESTS CELL 674 00:30:28,746 --> 00:30:31,915 CYCLE AND CANCER CAN'T PROPAGATE 675 00:30:31,915 --> 00:30:36,019 ANYMORE AND ORIGINAL IDEA AND 676 00:30:36,019 --> 00:30:40,023 REPURPOSING MOUSE, SOMEHOW MOUSE 677 00:30:40,023 --> 00:30:42,326 IS FIXED WE DON'T KNOW 678 00:30:42,326 --> 00:30:43,527 MOLECULARLY WHAT IS HAPPENING 679 00:30:43,527 --> 00:30:45,763 AND SET UP PIPELINE SO MODIFIED 680 00:30:45,763 --> 00:30:47,264 OR ADVANCED VERSION COMPARED TO 681 00:30:47,264 --> 00:30:49,733 OUR SHOWCASE STUDY. 682 00:30:49,733 --> 00:30:54,271 YOU SEE HERE WE PUT MORE LIGHT 683 00:30:54,271 --> 00:30:55,572 MICROSCOPIC EMPHASIS ON STUDY TO 684 00:30:55,572 --> 00:30:59,476 HAVE A LOOK WHAT IS GOING ON AND 685 00:30:59,476 --> 00:31:02,713 WE HAVE THIS EM GRADE PUTTING 686 00:31:02,713 --> 00:31:05,849 NEUROTISSUE HERE TO LET AXON TO 687 00:31:05,849 --> 00:31:06,350 GROW. 688 00:31:06,350 --> 00:31:13,657 ONE BIG THING WE DO IS AXOTOMY 689 00:31:13,657 --> 00:31:16,660 AND WE CAN IMPLEMENT TO TOP OF 690 00:31:16,660 --> 00:31:19,129 AXON ON EM GRADES I WILL SHOW 691 00:31:19,129 --> 00:31:21,665 YOU LATER TO OBSERVE WHAT IS THE 692 00:31:21,665 --> 00:31:25,869 RESPONSE ALSO ON CRYO-EM. 693 00:31:25,869 --> 00:31:34,878 SO, I SHOW YOU HOW WE ARE DOING. 694 00:31:34,878 --> 00:31:36,346 THIS IS THE SCOMPLANT AND 695 00:31:36,346 --> 00:31:39,116 NEURONS COMING OUT FROM THE 696 00:31:39,116 --> 00:31:41,118 TISSUE AND THEY ARE AXONS THAT 697 00:31:41,118 --> 00:31:42,419 ARE USEFUL FOR US. 698 00:31:42,419 --> 00:31:44,655 WE HAVE GOOD CONTROL OF IT AND 699 00:31:44,655 --> 00:31:47,591 WE CAN IMPLEMENT THAT ON TO EM 700 00:31:47,591 --> 00:31:49,126 GRADE YOU CAN SEE HERE. 701 00:31:49,126 --> 00:31:51,562 THIS IS A TISSUE AND AXONS AND 702 00:31:51,562 --> 00:31:54,264 WE CAN GROW AXONS UP TO SOME 703 00:31:54,264 --> 00:31:56,467 DAYS AND AROUND 8 DAYS IT 704 00:31:56,467 --> 00:31:57,868 DOESN'T GROW ANYMORE AND IS TIME 705 00:31:57,868 --> 00:32:00,471 WE CAN TEST REGENERATION. 706 00:32:00,471 --> 00:32:02,973 SO, THIS IS THE LIGHT MICROSCOPE 707 00:32:02,973 --> 00:32:05,008 STUDY TO SHOW YOU WHAT IS 708 00:32:05,008 --> 00:32:07,277 HAPPENING AND SO IN CELLULAR 709 00:32:07,277 --> 00:32:10,481 LEVEL WE CAN SEE ALSO I DON'T 710 00:32:10,481 --> 00:32:12,916 KNOW IF YOU NOTICE THAT CUT AXON 711 00:32:12,916 --> 00:32:18,188 WAS REGENERATED IN PRESENCE OF 712 00:32:18,188 --> 00:32:19,790 EPOB. IN PRESENCE OF IT IT 713 00:32:19,790 --> 00:32:21,558 SHRINKS TO MAKE IT EASIER LIKE 714 00:32:21,558 --> 00:32:24,294 THIS IT GROWS BACK THIS EPOB AND 715 00:32:24,294 --> 00:32:27,498 SHRINKS EPOB AND WE CAN 716 00:32:27,498 --> 00:32:29,800 REPRODUCE WHAT STUDIES SHOWS IN 717 00:32:29,800 --> 00:32:31,502 CELLS THAT IS QUITE NICE AND WE 718 00:32:31,502 --> 00:32:35,072 CAN ALSO QUANTIFY THAT. 719 00:32:35,072 --> 00:32:37,474 NOW LET'S HAVE A LOOK AT SINGLE 720 00:32:37,474 --> 00:32:40,477 AXON CAREFULLY AND YOU SEE HERE 721 00:32:40,477 --> 00:32:42,679 THIS IS THERE. IT CAN BE QUICK. 722 00:32:42,679 --> 00:32:46,884 IT JUMPED AND IT IS STARTING TO 723 00:32:46,884 --> 00:32:47,451 REGENERATE ALREADY. 724 00:32:47,451 --> 00:32:50,220 SO LET'S JUST MAKE A SNAPSHOT OF 725 00:32:50,220 --> 00:32:53,991 IT AROUND 50 MINUTES OR ONE HOUR 726 00:32:53,991 --> 00:32:56,460 YOU ACTUALLY DON'T SEE THE 727 00:32:56,460 --> 00:32:59,997 TYPICAL GROWTH OF THE GROWTH 728 00:32:59,997 --> 00:33:00,364 CON. 729 00:33:00,364 --> 00:33:04,735 I DO ACTUALLY SEE THE GROWTH 730 00:33:04,735 --> 00:33:04,935 CON. 731 00:33:04,935 --> 00:33:07,304 WE KNOW IT BETWEEN ZERO AND 50 732 00:33:07,304 --> 00:33:11,041 MINUTES OF REGENERATION IT IS 733 00:33:11,041 --> 00:33:12,309 REGENERATING WITHOUT FAMOUS 734 00:33:12,309 --> 00:33:13,310 GROWTH CON. 735 00:33:13,310 --> 00:33:15,145 WHAT DOES IT MEAN? 736 00:33:15,145 --> 00:33:20,117 WE WONDER HOW THIS CAN 737 00:33:20,117 --> 00:33:21,051 REGENERATE WITHOUT IMPORTANT 738 00:33:21,051 --> 00:33:23,353 GROWTH CON FORMATION. 739 00:33:23,353 --> 00:33:26,790 YOU CAN ALSO SEE IT HERE IN THE 740 00:33:26,790 --> 00:33:30,394 PRESENCE OF THE EPOB AFTER ONE 741 00:33:30,394 --> 00:33:30,961 HOUR. 742 00:33:30,961 --> 00:33:34,631 YOU GET IT REGENERATED BUT DON'T 743 00:33:34,631 --> 00:33:36,667 SEE GROWTH COM. 744 00:33:36,667 --> 00:33:39,636 THIS IS HYPOTHESIZED THIS IS A 745 00:33:39,636 --> 00:33:44,041 MICROTUBULE DOING THE PUSH TO 746 00:33:44,041 --> 00:33:46,410 REGENERATE REGENERATING AXON AND 747 00:33:46,410 --> 00:33:48,378 THIS IS IT COMPROMISED OR NOT? 748 00:33:48,378 --> 00:33:50,714 HOW IS MEMBRANE LOOKING LIKE? 749 00:33:50,714 --> 00:33:54,384 WITH THAT WE ACTUALLY ENDED UP 750 00:33:54,384 --> 00:33:58,855 MEASURING MEMBRANE TENSION USING 751 00:33:58,855 --> 00:34:00,724 MEMBRANE TENSION DIKON FLIPPER 752 00:34:00,724 --> 00:34:04,161 AND TIP OF REGENERATION AXON 753 00:34:04,161 --> 00:34:06,296 INDICATING THIS IS VERY TENSE 754 00:34:06,296 --> 00:34:07,831 AND MEMBRANE IS TENSE AND NOT 755 00:34:07,831 --> 00:34:09,600 NORMAL MEMBRANE. 756 00:34:09,600 --> 00:34:09,933 SORRY. 757 00:34:09,933 --> 00:34:12,035 IN THE MEANTIME AFTER SOME OF US 758 00:34:12,035 --> 00:34:13,904 AFTER FOUR HOURS YOU SEE TENSION 759 00:34:13,904 --> 00:34:16,306 TO BE NORMALIZED AND THIS MEANS 760 00:34:16,306 --> 00:34:18,675 AT BEGINNING THERE IS A VERY, 761 00:34:18,675 --> 00:34:21,945 VERY FORCED PUSH FOR THE AXON TO 762 00:34:21,945 --> 00:34:24,247 REGENERATE AND AT SOME POINT 763 00:34:24,247 --> 00:34:27,217 GROWTH CON COMES BACK AND IS 764 00:34:27,217 --> 00:34:28,885 NORMAL REGENERATION AND LET'S 765 00:34:28,885 --> 00:34:33,223 HAVE A LOOK AT THAT ON CRYO-EM. 766 00:34:33,223 --> 00:34:35,292 YOU CAN SEE HERE THIS IS THE 767 00:34:35,292 --> 00:34:36,026 NEURON. 768 00:34:36,026 --> 00:34:38,161 THIS IS THE EM GRADE. 769 00:34:38,161 --> 00:34:40,030 THIS IS THOUGHT, EACH OF THE 770 00:34:40,030 --> 00:34:42,432 THOUGHT HAS DIAMETER OF A 771 00:34:42,432 --> 00:34:45,002 MICROMETER THAT WE TYPICALLY USE 772 00:34:45,002 --> 00:34:47,137 FOR THE IMAGE ANALYSIS OR 773 00:34:47,137 --> 00:34:51,274 OBSERVATION OF THE AREA AND THIS 774 00:34:51,274 --> 00:34:52,542 INSITU WE DON'T USUALLY CARE 775 00:34:52,542 --> 00:34:54,645 ABOUT THE HOLE BUT SEE WHAT WE 776 00:34:54,645 --> 00:34:54,945 SEE. 777 00:34:54,945 --> 00:34:57,581 NOW THAT WE PERFORMED AXE ON MY 778 00:34:57,581 --> 00:35:00,017 ON TO EM GRADE YOU SEE HERE WE 779 00:35:00,017 --> 00:35:02,886 ARE CLEANING DEBRIS SO WE DON'T 780 00:35:02,886 --> 00:35:03,620 GET CONFUSED. 781 00:35:03,620 --> 00:35:04,855 NOW LET'S JUST HAVE A LOOK AT 782 00:35:04,855 --> 00:35:08,392 WHAT IS HAPPENING THERE. 783 00:35:08,392 --> 00:35:11,028 SO TO CONFOUND REGENERATION WHAT 784 00:35:11,028 --> 00:35:13,497 WOULD HAPPEN ON EM GRADE YOU SEE 785 00:35:13,497 --> 00:35:15,666 COLORED AXON THAT GOT 786 00:35:15,666 --> 00:35:17,267 REGENERATED AND WE CAN REALLY 787 00:35:17,267 --> 00:35:20,003 FOLLOW REGENERATION AND HAVING A 788 00:35:20,003 --> 00:35:21,471 LOOK AFTER ONE HOUR. 789 00:35:21,471 --> 00:35:23,173 SO AFTER ONE HOUR HOW IT LOOKS 790 00:35:23,173 --> 00:35:24,107 IS LIKE THIS. 791 00:35:24,107 --> 00:35:30,380 THIS IS A TISSUE THAT IS THE 792 00:35:30,380 --> 00:35:32,015 EXPLANT AXONS. 793 00:35:32,015 --> 00:35:36,019 YOU SEE TWO AXONS WE DID 794 00:35:36,019 --> 00:35:39,956 PERFORMANCE OF AXONOMY AND 795 00:35:39,956 --> 00:35:40,791 REGENERATION YOU SEE HERE OF 796 00:35:40,791 --> 00:35:43,193 STUFF HAPPENING AND LOOKING A 797 00:35:43,193 --> 00:35:44,761 LITTLE MORE, WITH THIS I'M 798 00:35:44,761 --> 00:35:46,797 PLEASED TO NOT GET ALERTED. 799 00:35:46,797 --> 00:35:49,933 WE DO NOT HAVE THE MEMBRANE 800 00:35:49,933 --> 00:35:51,902 DENSITY WHICH WE DON'T. IT IS 801 00:35:51,902 --> 00:35:53,970 DUE TO TENSION. 802 00:35:53,970 --> 00:35:56,473 SO, DURING THE FREEZING PROCESS, 803 00:35:56,473 --> 00:35:59,810 MEMBRANE DID NOT SURVIVE. 804 00:35:59,810 --> 00:36:01,678 WHAT WAS ADVANTAGE IN 805 00:36:01,678 --> 00:36:03,313 [INDISCERNIBLE] AND WE GET A 806 00:36:03,313 --> 00:36:06,149 VERY GOOD SIGNAL RATIO TO ALLOW 807 00:36:06,149 --> 00:36:08,585 US TO DO HIGH RESOLUTION 808 00:36:08,585 --> 00:36:08,852 ANALYSIS. 809 00:36:08,852 --> 00:36:10,320 LET'S HAVE A LOOK. 810 00:36:10,320 --> 00:36:12,689 YOU CAN SEE HERE THIS IS NORMAL 811 00:36:12,689 --> 00:36:15,092 AXON AND YOU SEE THIS CUT SIDE 812 00:36:15,092 --> 00:36:17,494 AND SEE CRAZY AMOUNT OF THE 813 00:36:17,494 --> 00:36:19,396 REGENERATION TO BE STARTING TO 814 00:36:19,396 --> 00:36:19,629 HAPPEN. 815 00:36:19,629 --> 00:36:23,633 YOU SEE LOTS OF MICROTUBULE AND 816 00:36:23,633 --> 00:36:25,502 VESICLES TRANSPORTED. 817 00:36:25,502 --> 00:36:28,038 WE DON'T KNOW MUCH ABOUT BASIC 818 00:36:28,038 --> 00:36:29,439 TRANSPORT ISSUE YET. WE ARE 819 00:36:29,439 --> 00:36:31,608 DOING IT RIGHT NOW AND KNOW LOTS 820 00:36:31,608 --> 00:36:35,011 OF STUFF IS TRANSPORTED ALONG 821 00:36:35,011 --> 00:36:36,747 MICROTUBULE THAT IS SHOOTED OUT 822 00:36:36,747 --> 00:36:40,016 AND THIS LENGTHWISE IS AROUND 20 823 00:36:40,016 --> 00:36:41,785 MICROMETERS QUITE A LOT OF 824 00:36:41,785 --> 00:36:43,553 ELONGATION OF CELL THAT IS 825 00:36:43,553 --> 00:36:45,689 HAPPENING WITHIN ONE HOUR. 826 00:36:45,689 --> 00:36:47,557 NOW, IN ORDER TO UNDERSTAND HOW 827 00:36:47,557 --> 00:36:50,527 AND WHAT IS THE MOLECULAR BASIS 828 00:36:50,527 --> 00:36:52,729 OF THIS SUPER SHOOTING OUT 829 00:36:52,729 --> 00:36:55,599 EVENT, WE DECIDED TO DO LARGE 830 00:36:55,599 --> 00:36:58,101 SCALE AXOTOMY. THAT MEANS YOU 831 00:36:58,101 --> 00:37:00,036 HAVE LOTS OF AXONS HERE AND WE 832 00:37:00,036 --> 00:37:02,005 CAN ACTUALLY CUT THAT AND WE CAN 833 00:37:02,005 --> 00:37:06,409 OBSERVE THE ENTIRE AREA HERE AS 834 00:37:06,409 --> 00:37:08,678 THINKING THAT THE TENSION OF THE 835 00:37:08,678 --> 00:37:10,914 MEMBRANE IS ALL APPLIED SO THAT 836 00:37:10,914 --> 00:37:15,152 IF YOU DO CRYO-EM ALL OVER HERE 837 00:37:15,152 --> 00:37:17,087 DON'T HAVE THE MEMBRANELESS LESS 838 00:37:17,087 --> 00:37:19,055 COMPONENTS AND WE CAN DO 839 00:37:19,055 --> 00:37:23,160 ANALYSIS AND WE COLLECTED DATA 840 00:37:23,160 --> 00:37:25,028 OF 2000 IMAGES AND HAVE A LOOK 841 00:37:25,028 --> 00:37:27,864 AT IT AND DO SPS [INDISCERNIBLE] 842 00:37:27,864 --> 00:37:29,199 THAT GIVES HIGH RESOLUTION 843 00:37:29,199 --> 00:37:31,635 INFORMATION AND WE CAN DO 844 00:37:31,635 --> 00:37:34,304 TOMOGRAPHY TO GET CONTEXT AND 845 00:37:34,304 --> 00:37:36,039 FOR TODAY'S TALK I WILL TALK 846 00:37:36,039 --> 00:37:38,275 ABOUT SPA VERSION OF IT. 847 00:37:38,275 --> 00:37:41,111 BY COLLECTING SO MANY DATA, WE 848 00:37:41,111 --> 00:37:44,014 COULD GET 3-D RECONSTRUCTION OF 849 00:37:44,014 --> 00:37:46,183 THE MICROTUBULE AT REGENERATING 850 00:37:46,183 --> 00:37:50,487 AXON. YOU SEE HERE THIS IS 851 00:37:50,487 --> 00:37:52,923 FILAMENT THAT IS PROTEIN THAT IS 852 00:37:52,923 --> 00:37:56,026 IMPORTANT FOR MICROTUBULE IF YOU 853 00:37:56,026 --> 00:37:57,694 [INDISCERNIBLE] IT WILL BE AND 854 00:37:57,694 --> 00:38:02,132 IS NOT REALLY CONVENIENT FOR 855 00:38:02,132 --> 00:38:02,365 CELLS. 856 00:38:02,365 --> 00:38:06,369 YOU CAN SEE [INDISCERNIBLE] THAT 857 00:38:06,369 --> 00:38:10,273 IS A PROTEIN THAT IS MAKING 858 00:38:10,273 --> 00:38:11,708 [INDISCERNIBLE] OF MICROTUBULE 859 00:38:11,708 --> 00:38:13,877 TO BE CERTAIN PROTEIN ALL OF 860 00:38:13,877 --> 00:38:16,680 THEM 1 THOUSAND 6 HUNDRED 63 861 00:38:16,680 --> 00:38:18,315 MICROTUBULES THAT WE OBSERVED. 862 00:38:18,315 --> 00:38:21,618 WE CAN SEE AND GO ANALYSIS OF 863 00:38:21,618 --> 00:38:24,020 DIRECTIONALITY TO SHOW THEY ARE 864 00:38:24,020 --> 00:38:27,290 ALL HAVING SAME DIRECTIONALITY 865 00:38:27,290 --> 00:38:28,425 WHICH IS AN IMPORTANT IDENTITY 866 00:38:28,425 --> 00:38:30,327 FOR THE AXON. 867 00:38:30,327 --> 00:38:32,529 DENDRITE MICROTUBULES ARE 868 00:38:32,529 --> 00:38:35,365 SWAPPED AROUND. 869 00:38:35,365 --> 00:38:36,032 SO, SORRY. 870 00:38:36,032 --> 00:38:38,635 SO THIS REALLY INDICATES THAT 871 00:38:38,635 --> 00:38:41,671 MICROTUBULES ARE READY TO BECOME 872 00:38:41,671 --> 00:38:45,141 THE REAR AXON THAT WE ARE 873 00:38:45,141 --> 00:38:45,775 REGENERATING. 874 00:38:45,775 --> 00:38:50,380 THIS RESOLUTION, WE CAN ALSO SEE 875 00:38:50,380 --> 00:38:55,719 AN INTERDIMER DISTANCES 83.3 876 00:38:55,719 --> 00:38:56,786 [INDISCERNIBLE]IN SITU AND IN 877 00:38:56,786 --> 00:38:59,022 VITRO WE CAN SEE IT AND CONTROL 878 00:38:59,022 --> 00:39:01,658 IT IN STABLE CONDITION WE KNOW 879 00:39:01,658 --> 00:39:04,027 IT HAS 84 [INDISCERNIBLE] AND 880 00:39:04,027 --> 00:39:05,528 MICROTUBULE WHEN IT IS NOT 881 00:39:05,528 --> 00:39:09,766 STABLE IS 81.8 ANGSTROM THIS 882 00:39:09,766 --> 00:39:11,534 INDICATES MICROTUBULES AT 883 00:39:11,534 --> 00:39:12,903 REGENERATING SITE OF THE AXON IS 884 00:39:12,903 --> 00:39:17,941 A STABILIZED FORM OF THE AXON OR 885 00:39:17,941 --> 00:39:18,408 MICROTUBULES. 886 00:39:18,408 --> 00:39:20,510 WE WONDER THAT THERE MUST BE 887 00:39:20,510 --> 00:39:22,746 EPOB THAT IS ATTACHED TO 888 00:39:22,746 --> 00:39:25,548 STABILIZE MICROTUBULE AND DIDN'T 889 00:39:25,548 --> 00:39:27,484 SEE IT WHEN WE HAVE HIGH 890 00:39:27,484 --> 00:39:29,219 THRESHOLD OF THE DENSITY AND 891 00:39:29,219 --> 00:39:31,488 WHEN WE HAVE -- WE CHANGE 892 00:39:31,488 --> 00:39:33,723 THRESHOLD WE START TO SEE EPOB. 893 00:39:33,723 --> 00:39:35,926 THIS IS FIRST TIME WE CAN REALLY 894 00:39:35,926 --> 00:39:39,229 OBSERVE THE DRUG THAT IS ACTING 895 00:39:39,229 --> 00:39:42,933 ON TO THE MICROTUBULE IN 896 00:39:42,933 --> 00:39:44,834 REGENERATING CELL AND WE CAN SAY 897 00:39:44,834 --> 00:39:48,038 THAT DRUG WE ADDED IS 898 00:39:48,038 --> 00:39:49,272 PHARMACEUTICALLY RELEVANT 899 00:39:49,272 --> 00:39:51,441 CONCENTRATION THAT IS ONLY ONE 900 00:39:51,441 --> 00:39:53,977 NANOMOLE AR OR MICROTUBULES 901 00:39:53,977 --> 00:39:56,613 DENSITY IS IN MICROMOLARS 902 00:39:56,613 --> 00:39:57,914 THOUSANDS OF MAGNITUDE LOWER 903 00:39:57,914 --> 00:40:02,118 AMOUNT OF THE DRUG THAT IS ADDED 904 00:40:02,118 --> 00:40:04,454 COMPARED TO TUBULIN AND THIS 905 00:40:04,454 --> 00:40:05,388 DENSITY HERE AND CONCENTRATION 906 00:40:05,388 --> 00:40:08,725 OF THE DRUG IS HAPPENING AT 907 00:40:08,725 --> 00:40:11,761 REGENERATING SITE OF THE AXON. 908 00:40:11,761 --> 00:40:14,397 OKAY. NOW WE ARE AWARE WHERE IS 909 00:40:14,397 --> 00:40:15,899 MICROTUBULE COMING FROM? WE 910 00:40:15,899 --> 00:40:18,401 HAVE THREE HYPOTHESIS TO ASK AND 911 00:40:18,401 --> 00:40:21,571 GOING ONE-BY-ONE AND FIRST 912 00:40:21,571 --> 00:40:24,007 HYPOTHESIS THAT TUBULIN IS 913 00:40:24,007 --> 00:40:26,076 SYNTHESIZED AT REGENERATING SITE 914 00:40:26,076 --> 00:40:29,579 AND WE THOUGHT SO BECAUSE WE 915 00:40:29,579 --> 00:40:32,015 SOMETHING LIKE THAT ACTING AND 916 00:40:32,015 --> 00:40:34,584 THIS IS AXON AND AXON BRANCH AND 917 00:40:34,584 --> 00:40:37,120 YOU SEE RIBOSOMES HERE AND SEE 918 00:40:37,120 --> 00:40:38,955 ACTIN THAT IS COMING OUT FROM 919 00:40:38,955 --> 00:40:40,423 THE RIBOSOME. 920 00:40:40,423 --> 00:40:42,258 WE HAVE REGENERATION. 921 00:40:42,258 --> 00:40:47,163 SORRY AXON THAT IS BRANCHING 922 00:40:47,163 --> 00:40:52,035 GENERATION WE WOULD SEE SINNIS 923 00:40:52,035 --> 00:40:54,471 OF ACTIN AND REGENERATION SIDE 924 00:40:54,471 --> 00:40:56,339 AND DON'T SEE MUCH INTERACTION 925 00:40:56,339 --> 00:40:58,441 WE HAVE NOT SEEN IT SO FAR AND 926 00:40:58,441 --> 00:41:00,010 STILL LOOKING AROUND AND HAVEN'T 927 00:41:00,010 --> 00:41:02,379 FOUND IT SO FAR AND IS NOT A 928 00:41:02,379 --> 00:41:04,614 PRIME MECHANISM TO PRODUCE 929 00:41:04,614 --> 00:41:07,917 TUBING COMING TO REGENERATION OF 930 00:41:07,917 --> 00:41:09,052 THE AXON. 931 00:41:09,052 --> 00:41:12,856 SO, NEXT HYPOTHESIS THAT WE 932 00:41:12,856 --> 00:41:15,825 WONDER, SORRY. 933 00:41:15,825 --> 00:41:18,962 I SHOWED THIS ONE FIRST. 934 00:41:18,962 --> 00:41:21,031 THIS IS [INDISCERNIBLE] OF 935 00:41:21,031 --> 00:41:22,499 TUBULING AND REGENERATION THAT 936 00:41:22,499 --> 00:41:25,702 IS HAPPENING AND CLUSTER OF 937 00:41:25,702 --> 00:41:27,637 TUBULING OR MICROTUBULE IS 938 00:41:27,637 --> 00:41:29,506 SHATTERED TO TIP OF REGENERATING 939 00:41:29,506 --> 00:41:33,009 AXON AND MEANS THERE IS SOME 940 00:41:33,009 --> 00:41:37,247 SORT OF A GENERATION OF THE 941 00:41:37,247 --> 00:41:41,885 TUBULIN OR MICROTUBE YULE AND 942 00:41:41,885 --> 00:41:43,653 PRECUT SIDE AND IT GETS 943 00:41:43,653 --> 00:41:47,690 SHATTERED IS IT MICROTUBULE OR 944 00:41:47,690 --> 00:41:52,028 TUBULIN? IF MICROTUBULE WE KNOW 945 00:41:52,028 --> 00:41:55,799 NORMAL MICROTUBULE IN NEURON 946 00:41:55,799 --> 00:41:58,835 CONTAINS MIPS THAT ARE USUALLY 947 00:41:58,835 --> 00:42:00,770 THERE WHEN MICROTUBULES ARE 948 00:42:00,770 --> 00:42:02,906 MATURE AND LOOKING AT 949 00:42:02,906 --> 00:42:04,941 REGENERATING AXON MICROTUBULES 950 00:42:04,941 --> 00:42:08,745 HAVE MUCH LESS OF MIPS INSIDE 951 00:42:08,745 --> 00:42:11,548 AND MEANING THEY ARE YOUNG 952 00:42:11,548 --> 00:42:14,284 MICROTUBULES NOT MATURED YET TO 953 00:42:14,284 --> 00:42:16,152 CONTAIN INTERLUMIN PARTICLES WE 954 00:42:16,152 --> 00:42:19,322 CAN MEASURE IT AND YOU SEE HERE 955 00:42:19,322 --> 00:42:21,124 MICROTUBULES ARE MUCH LESS 956 00:42:21,124 --> 00:42:23,393 CROWDED COMPARED TO THE NORMAL 957 00:42:23,393 --> 00:42:23,793 CONTROL. 958 00:42:23,793 --> 00:42:26,062 THIS INDICATES THAT MICROTUBULES 959 00:42:26,062 --> 00:42:31,367 ARE NOT CARRIED FROM THE SOMA OR 960 00:42:31,367 --> 00:42:34,637 UPSTREAM OF THE AXON BUT IS 961 00:42:34,637 --> 00:42:37,040 POLYMERIZED AT THE REGENERATING 962 00:42:37,040 --> 00:42:38,708 SITE AND EXTREME EXAMPLE, YOU 963 00:42:38,708 --> 00:42:42,278 CAN SEE HERE THEY ARE TIP OF 964 00:42:42,278 --> 00:42:43,847 REGENERATING SITE AND 965 00:42:43,847 --> 00:42:46,416 MICROTUBULES DON'T HAVE MIPS. 966 00:42:46,416 --> 00:42:48,651 THEY ARE FRESHLY MADE 967 00:42:48,651 --> 00:42:50,186 MICROTUBULES AT THE TIP. 968 00:42:50,186 --> 00:42:54,224 NOW, HAVING THAT IN OUR MIND WE 969 00:42:54,224 --> 00:42:56,926 LOOK INTO POLYMERIZATION PROCESS 970 00:42:56,926 --> 00:43:01,498 OF MICROTUBULE AND TRIED TO FIND 971 00:43:01,498 --> 00:43:02,265 IT. 972 00:43:02,265 --> 00:43:04,868 HERE IS THE BASIC INFORMATION 973 00:43:04,868 --> 00:43:06,236 THAT IS SHOWN ALREADY AND SHOWN 974 00:43:06,236 --> 00:43:09,706 WHEN MICROTUBULE IS POLYMERIZED 975 00:43:09,706 --> 00:43:12,775 AND SEE IN THE BACKGROUND 976 00:43:12,775 --> 00:43:16,779 OLIGGANDS IN MICROTUBULE AND IN 977 00:43:16,779 --> 00:43:20,116 THIS PRIOR CRYO-EM STUDY IN 978 00:43:20,116 --> 00:43:23,386 VITRO YOU SEE THIS SPIRAL LIKE 979 00:43:23,386 --> 00:43:26,523 OLIGGAND THAT GETS ATTACHED TO 980 00:43:26,523 --> 00:43:28,324 THE MICROTUBULE AND STRAIGHTENED 981 00:43:28,324 --> 00:43:30,326 INTO THIS AND IT GETS 982 00:43:30,326 --> 00:43:32,028 INCORPORATED AND WE LOOK INTO 983 00:43:32,028 --> 00:43:35,798 THAT IN OUR OWN TOMMO GRAMS OF 984 00:43:35,798 --> 00:43:36,432 REGENERATING AXON. 985 00:43:36,432 --> 00:43:39,302 YOU WOULD SEE HERE THERE IS A 986 00:43:39,302 --> 00:43:41,371 BIG CLUSTER OF OLIGGANDS THAT 987 00:43:41,371 --> 00:43:45,642 ARE LOOKING SIMILAR TO WHAT IS 988 00:43:45,642 --> 00:43:49,512 SHOWN AS IN VITRO TUBULE 989 00:43:49,512 --> 00:43:50,813 OLIGGANDS HERE IS SNAPSHOT YOU 990 00:43:50,813 --> 00:43:52,015 CAN SEE MORE. 991 00:43:52,015 --> 00:43:56,019 WE MEASURE DIAMETER OR CURVATURE 992 00:43:56,019 --> 00:44:01,391 OF OLIGOMER THAT IS SHOWN IN 993 00:44:01,391 --> 00:44:04,861 VITRO AND WE DO SEGMENTATION TO 994 00:44:04,861 --> 00:44:07,263 SEE AND IT SHOWS SPIRAL 995 00:44:07,263 --> 00:44:10,066 FORMATION AND DETECT SIZE OF 996 00:44:10,066 --> 00:44:14,237 MONOMER THAT IS INCORPORATED 997 00:44:14,237 --> 00:44:15,939 INTO SPIRALS. 998 00:44:15,939 --> 00:44:19,142 YOU SEE HERE THIS STAND OUT 999 00:44:19,142 --> 00:44:21,978 NANOMETER OR.5 NANOMETER THAT 1000 00:44:21,978 --> 00:44:26,716 FITS IF IN UNIT THAT YOU SEE IN 1001 00:44:26,716 --> 00:44:30,086 MICROTUBULE IN TOMMO GRAM AND 1002 00:44:30,086 --> 00:44:32,021 TOMOGRAPHY RESOLUTION IS GOOD 1003 00:44:32,021 --> 00:44:34,390 ENOUGH TO DISTINGUISH SIZE OF 1004 00:44:34,390 --> 00:44:38,161 ACTING MONOMER AND TUBULIN 1005 00:44:38,161 --> 00:44:40,063 MONOMER AND YOU CAN SEE ALL 1006 00:44:40,063 --> 00:44:44,834 TOGETHER THIS IS TUBULIN 1007 00:44:44,834 --> 00:44:45,301 MONOMER. 1008 00:44:45,301 --> 00:44:47,136 TO GET FINAL CONFIRMATION OR 1009 00:44:47,136 --> 00:44:52,008 FINAL VALIDATION OF IT YOU SEE 1010 00:44:52,008 --> 00:44:53,943 OLIGOMERS OF TUBULIN WE ATTACH 1011 00:44:53,943 --> 00:44:57,180 TO END OF MICROTUBULE THAT ARE 1012 00:44:57,180 --> 00:44:58,781 POLYMERIZING TO CAPTURE SNAPSHOT 1013 00:44:58,781 --> 00:45:03,519 OF THE MICROTUBULES 1014 00:45:03,519 --> 00:45:04,854 POLYMERIZINGIN SEATU INTO 1015 00:45:04,854 --> 00:45:07,156 REGENERATING AXON AND THIS IS A 1016 00:45:07,156 --> 00:45:08,725 SIDENOTE WE HAVE BEEN FOLLOWING 1017 00:45:08,725 --> 00:45:10,026 UP FOR SOME TIME. 1018 00:45:10,026 --> 00:45:12,895 YOU SEE HERE THIS IS A 1019 00:45:12,895 --> 00:45:16,499 MICROTUBULE THAT GETS BRANCHED. 1020 00:45:16,499 --> 00:45:19,102 HERE YOU SEE THIS SHARING 1021 00:45:19,102 --> 00:45:20,103 MICROTUBE YULE BRANCH. 1022 00:45:20,103 --> 00:45:22,005 WHY WE CARE ABOUT THAT IS 1023 00:45:22,005 --> 00:45:24,073 BECAUSE WE -- SOME YEARS AGO WE 1024 00:45:24,073 --> 00:45:27,677 PUBLISHED A PAPER SHOWING THAT 1025 00:45:27,677 --> 00:45:29,145 MICROTUBE YULE CAN BRANCH OUT IN 1026 00:45:29,145 --> 00:45:31,047 VITRO AND IN PRESENCE OF THE 1027 00:45:31,047 --> 00:45:33,249 PROTEIN THAT IS ABUNDANT IN 1028 00:45:33,249 --> 00:45:33,983 NEURON. 1029 00:45:33,983 --> 00:45:35,752 WE LEFT IT AS AN OPEN QUESTION 1030 00:45:35,752 --> 00:45:40,023 IF THIS WOULD BE SOMETHING 1031 00:45:40,023 --> 00:45:42,659 RELEVANTIN SITU OR IN VIVO AND 1032 00:45:42,659 --> 00:45:46,262 DON'T THINK IT IS MAJOR 1033 00:45:46,262 --> 00:45:49,165 MECHANISM FOR THIS OR REMODELING 1034 00:45:49,165 --> 00:45:50,500 AND WONDERING IF YOU WOULD CATCH 1035 00:45:50,500 --> 00:45:52,702 IT AND SINCE WE ARE VERY HAPPY 1036 00:45:52,702 --> 00:45:56,139 WE CAUGHT IT WITH REGENERATING 1037 00:45:56,139 --> 00:45:57,473 AXON. 1038 00:45:57,473 --> 00:45:59,542 YUP. CONCLUSION OF THIS PART OF 1039 00:45:59,542 --> 00:46:04,681 THE STUDY IS IN THIS TUBULE 1040 00:46:04,681 --> 00:46:06,916 OLIGOMERS TRANSPORT IT IN AXON 1041 00:46:06,916 --> 00:46:10,219 AND REGENERATION SIDE. 1042 00:46:10,219 --> 00:46:13,256 NOW, I HAVE SHOWN UP TO ONE HOUR 1043 00:46:13,256 --> 00:46:16,025 AND 24 HOURS WHAT IS GOING ON 1044 00:46:16,025 --> 00:46:18,594 YOU SEE HERE THIS IS A CUT SIDE 1045 00:46:18,594 --> 00:46:21,831 AND NEURON MANAGED TO REGENERATE 1046 00:46:21,831 --> 00:46:23,166 QUITE WELL. 1047 00:46:23,166 --> 00:46:25,935 WE CAN ALSO TAKE TOPOGRAPHIC 1048 00:46:25,935 --> 00:46:29,205 SNAPSHOTS OF CERTAIN AREAS ALONG 1049 00:46:29,205 --> 00:46:30,707 REGENERATING AXON THAT SEEM TO 1050 00:46:30,707 --> 00:46:31,874 BE OKAY. 1051 00:46:31,874 --> 00:46:35,712 I MEAN, HEALTHY ENOUGH. 1052 00:46:35,712 --> 00:46:41,317 IT ALSO HAS THE GROWTH CONE 1053 00:46:41,317 --> 00:46:43,386 CAPPED ALREADY AND CAME BACK 1054 00:46:43,386 --> 00:46:43,619 NICELY. 1055 00:46:43,619 --> 00:46:45,755 NOW, CONCLUSION, AFTER ONE HOUR 1056 00:46:45,755 --> 00:46:49,492 OF THE AXOTOMY YOU SEE NO 1057 00:46:49,492 --> 00:46:52,562 TYPICAL GROWTH CONE I SHOW 1058 00:46:52,562 --> 00:46:54,430 MICROTUBE YULE GROW RAPIDLY THAT 1059 00:46:54,430 --> 00:46:58,301 IS ASSOCIATED WITH INCREASED 1060 00:46:58,301 --> 00:47:01,671 MEMBRANE TENSION AND INCREASE 1061 00:47:01,671 --> 00:47:04,640 REGENERATING SIDE AND TUBULE 1062 00:47:04,640 --> 00:47:07,243 OLIGOMERS UP TO 6 HOURS AND NEW 1063 00:47:07,243 --> 00:47:08,745 MEMBRANE AND LIPID THAT IS 1064 00:47:08,745 --> 00:47:10,413 STARTING WITH AXON GROWTH SEEING 1065 00:47:10,413 --> 00:47:15,651 BY CHANGE OF THE TENSION OF THE 1066 00:47:15,651 --> 00:47:15,918 MEMBRANE. 1067 00:47:15,918 --> 00:47:17,687 YEAH. THEN FINALLY SOME HOURS 1068 00:47:17,687 --> 00:47:20,490 AFTER WE SEE RECOVERY OF GROWTH 1069 00:47:20,490 --> 00:47:21,858 CONE THAT IS IMPORTANT THING FOR 1070 00:47:21,858 --> 00:47:25,728 THE AXON GUIDANCE ALSO THAT IS 1071 00:47:25,728 --> 00:47:28,464 GUIDED MOVEMENT OF THE GROWTH 1072 00:47:28,464 --> 00:47:32,034 AXON REGENERATING AXON AND 1073 00:47:32,034 --> 00:47:34,837 ECONVENIENT TULELY THIS COULD BE 1074 00:47:34,837 --> 00:47:37,373 CONNECTED HOPEFULLY INTO NEURAL 1075 00:47:37,373 --> 00:47:38,674 NETWORK AGAIN AND THIS IS A 1076 00:47:38,674 --> 00:47:40,009 FINAL NOTE OF MY TALK. 1077 00:47:40,009 --> 00:47:42,311 I SHOWED YOU ABOUT THE 1078 00:47:42,311 --> 00:47:45,381 RESOLUTION REVOLUTION OF WHAT IS 1079 00:47:45,381 --> 00:47:48,951 HAPPENING IN THE EM FIELD AND 1080 00:47:48,951 --> 00:47:50,753 IMAGING FIELD AND ALSO SOME OF 1081 00:47:50,753 --> 00:47:53,256 THE TWISTS AND ADDITIONAL 1082 00:47:53,256 --> 00:47:54,590 FACTORS AND DIMENSION THAT WOULD 1083 00:47:54,590 --> 00:47:57,126 BE GIVING US A LOT MORE OF 1084 00:47:57,126 --> 00:47:58,127 COMPREHENSIVE VIEW OF WHAT WE 1085 00:47:58,127 --> 00:48:01,197 ARE TRYING TO ADDRESS. 1086 00:48:01,197 --> 00:48:03,900 THIS WOULD BE NEW KIND OF BIG 1087 00:48:03,900 --> 00:48:05,635 PICTURE STUDY THAT WE TRIED TO 1088 00:48:05,635 --> 00:48:07,203 PASS THROUGH. 1089 00:48:07,203 --> 00:48:08,638 THAT IS THE DIRECTION THAT WE 1090 00:48:08,638 --> 00:48:10,373 HAVE GOING. 1091 00:48:10,373 --> 00:48:12,108 OKAY. SO WE START TO TRY TO 1092 00:48:12,108 --> 00:48:13,743 FINISH MY TALK AND I WANT TO 1093 00:48:13,743 --> 00:48:17,413 START BY ACKNOWLEDGING MY 1094 00:48:17,413 --> 00:48:19,916 COLLEAGUES AND STUDENTS AND 1095 00:48:19,916 --> 00:48:21,551 POST-DOCS FROM INSTITUTE OF 1096 00:48:21,551 --> 00:48:22,618 BIOCHEMISTRY AND THESE ARE 1097 00:48:22,618 --> 00:48:24,053 PEOPLE THAT STARTED UP TOGETHER 1098 00:48:24,053 --> 00:48:27,123 AND THEY DECIDED TO REMAIN IN 1099 00:48:27,123 --> 00:48:28,825 GERMANY AND MOVE ON ALREADY AND 1100 00:48:28,825 --> 00:48:32,028 THESE ARE THE PEOPLE THAT 1101 00:48:32,028 --> 00:48:34,197 ACTUALLY BRAVELY CAME WITH ME 1102 00:48:34,197 --> 00:48:36,566 FROM NIH AND THAT WAS REALLY 1103 00:48:36,566 --> 00:48:36,766 NICE. 1104 00:48:36,766 --> 00:48:39,168 WE COULD HAVE A KIND OF PRAITION 1105 00:48:39,168 --> 00:48:41,604 THAT WAS CONTINUING. 1106 00:48:41,604 --> 00:48:43,639 THESE ARE THE GRANTS I HAVE BEEN 1107 00:48:43,639 --> 00:48:45,608 GETTING IN GERMANY. 1108 00:48:45,608 --> 00:48:47,510 THIS IS QUITE GENEROUS SUPPORT I 1109 00:48:47,510 --> 00:48:50,112 WAS GETTING AND BEING A 1110 00:48:50,112 --> 00:48:51,581 NON-EUROPEAN BUT THEY SUPPORTED 1111 00:48:51,581 --> 00:48:52,381 ME. 1112 00:48:52,381 --> 00:48:53,749 THAT WAS VERY NICE. 1113 00:48:53,749 --> 00:48:56,519 HERE IS THE CURRENT NIH LAB. 1114 00:48:56,519 --> 00:48:58,187 SO WE ARE HAVING A VERY GOOD 1115 00:48:58,187 --> 00:49:05,194 TIME NOW AND TRYING TO MAKE SIN 1116 00:49:05,194 --> 00:49:05,595 EAR 1117 00:49:05,595 --> 00:49:07,730 EARTHY AND GOOD TEAM. I'M HAPPY 1118 00:49:07,730 --> 00:49:09,165 WITH MY NEIGHBOR. 1119 00:49:09,165 --> 00:49:11,901 WE HAVE A GOOD TIME TOGETHER AND 1120 00:49:11,901 --> 00:49:13,736 CO-FACILITY PARTICULARLY 1121 00:49:13,736 --> 00:49:16,005 MICROSCOPE CO-FACILITY AND THEY 1122 00:49:16,005 --> 00:49:17,707 ARE LIKE OUR EVERY DAY 1123 00:49:17,707 --> 00:49:19,675 DESTINATION TO GO IF YOU DON'T 1124 00:49:19,675 --> 00:49:21,143 FIND POSTER WE GO THERE TO FIND 1125 00:49:21,143 --> 00:49:23,212 THEM USUALLY AND FINALLY 1126 00:49:23,212 --> 00:49:25,147 COLLABORATORS AND YOU CAN SEE WE 1127 00:49:25,147 --> 00:49:27,250 HAVE LOTS OF COLLABORATION GOING 1128 00:49:27,250 --> 00:49:27,416 ON. 1129 00:49:27,416 --> 00:49:30,419 SO SOME OF THE COLLABORATORS AND 1130 00:49:30,419 --> 00:49:33,656 NEW COLLABORATORS AND WE HAVE 1131 00:49:33,656 --> 00:49:40,029 THEIR HELP AND THEIR INSIGHT AND 1132 00:49:40,029 --> 00:49:42,431 DIFFERENT PERSPECTIVE AND 1133 00:49:42,431 --> 00:49:44,166 ADDRESS WHAT WE START OUT TO 1134 00:49:44,166 --> 00:49:46,536 ACHIEVE AND THAT IS MY TALK AND 1135 00:49:46,536 --> 00:49:50,506 THANKS SO MUCH FOR YOUR 1136 00:49:50,506 --> 00:49:50,806 ATTENTION. 1137 00:49:50,806 --> 00:49:52,041 >> AUDIENCE: [APPLAUSE]. 1138 00:49:52,041 --> 00:49:58,581 >> WE WILL NOW OPEN UP TO 1139 00:49:58,581 --> 00:49:58,881 QUESTIONS. 1140 00:49:58,881 --> 00:50:00,049 IF ANYONE WOULD LIKE TO ANSWER 1141 00:50:00,049 --> 00:50:02,184 A QUESTION IF YOU COULD GO TO 1142 00:50:02,184 --> 00:50:02,985 THE MIC. 1143 00:50:02,985 --> 00:50:06,422 ONLINE ALSO IF YOU COULD USE THE 1144 00:50:06,422 --> 00:50:09,125 LIVE FEEDBACK I WILL PASS 1145 00:50:09,125 --> 00:50:13,329 QUESTIONS ON TO DR. MAZUNO. 1146 00:50:13,329 --> 00:50:18,434 >> OKAY. 1147 00:50:18,434 --> 00:50:28,911 >> VERY NICE OVERVIEW FROM 1148 00:50:33,015 --> 00:50:34,717 [INDISCERNIBLE] MY QUESTION IS 1149 00:50:34,717 --> 00:50:38,421 ABOUT EPLE B THE DRUG YOU SHOW 1150 00:50:38,421 --> 00:50:40,256 AND IN STRUCTURE YOU SHOW IT IS 1151 00:50:40,256 --> 00:50:44,026 BIND DIRECTLY TO TUBULING ON 1152 00:50:44,026 --> 00:50:45,561 TUBULING FILAMENT FORM AND ON 1153 00:50:45,561 --> 00:50:47,063 FUNCTION YOU MENTIONED IT 1154 00:50:47,063 --> 00:50:51,601 ACTUALLY HELPED TRANSPORTING OF 1155 00:50:51,601 --> 00:50:53,836 TUBULIN OLICCO MERS AND THAT 1156 00:50:53,836 --> 00:50:55,705 DOESN'T SEEM TO BE EXACTLY IN 1157 00:50:55,705 --> 00:50:57,273 THE STRUCTURE THAT DOESN'T 1158 00:50:57,273 --> 00:50:57,807 EXPLAIN THE FUNCTION. 1159 00:50:57,807 --> 00:50:59,875 >> RIGHT. RIGHT. WHAT WE THINK 1160 00:50:59,875 --> 00:51:01,510 IS HAPPENING IS THAT IN THE 1161 00:51:01,510 --> 00:51:04,180 PRESENCE OF EPOB AND THEN WHEN 1162 00:51:04,180 --> 00:51:07,550 THE POLYMER -- DEPOLYMERIZATION 1163 00:51:07,550 --> 00:51:10,086 OF THE MICROTUBULE TO BE 1164 00:51:10,086 --> 00:51:12,688 HAPPENING AND IN VITRO PEOPLE 1165 00:51:12,688 --> 00:51:14,023 HAVE SHOWN ALREADY THAT IT 1166 00:51:14,023 --> 00:51:16,626 BECOMES PIECES OF OLIGOMERS AND 1167 00:51:16,626 --> 00:51:19,829 WHAT WE THINK IS HAPPENING IS 1168 00:51:19,829 --> 00:51:22,665 THAT MAYBE IN E POB AND 1169 00:51:22,665 --> 00:51:25,101 MICROTUBULES THAT TRY TO DO 1170 00:51:25,101 --> 00:51:28,004 DYNAMIC INSTABILITY AND TRY TO 1171 00:51:28,004 --> 00:51:28,838 DO DEPOLYMERIZATION HERE AND 1172 00:51:28,838 --> 00:51:33,409 THERE AND PASSIONATELY 1173 00:51:33,409 --> 00:51:34,910 DEPOLYMERIZE MICROTUBULE 1174 00:51:34,910 --> 00:51:37,813 PRODUCING OLIGOMERS AND WHAT WE 1175 00:51:37,813 --> 00:51:40,016 ARE SEEING AS DOT FORMING WHEN 1176 00:51:40,016 --> 00:51:41,484 FORMING EPOB. 1177 00:51:41,484 --> 00:51:43,386 >> RIGHT. THEN YOU AT THE 1178 00:51:43,386 --> 00:51:45,821 BEGINNING YOU MENTIONED EPOB WAS 1179 00:51:45,821 --> 00:51:48,624 DEVELOPED AS AN ANTICANCER DRUG. 1180 00:51:48,624 --> 00:51:48,891 >> RIGHT. 1181 00:51:48,891 --> 00:51:51,627 >> HOW DID IT WORK AS ANTICANCER 1182 00:51:51,627 --> 00:51:54,430 DRUG IN TERMS OF 1183 00:51:54,430 --> 00:51:56,032 DEPOLYMERIZATION OF MICROTUBULE 1184 00:51:56,032 --> 00:51:57,299 OR MOVING IT. 1185 00:51:57,299 --> 00:52:00,102 >> YEAH. SO I THINK THE DESIGN 1186 00:52:00,102 --> 00:52:04,507 IS THAT IT ARREST THE -- SO 1187 00:52:04,507 --> 00:52:07,076 MICROTUBULE NEED TO REMODEL WHEN 1188 00:52:07,076 --> 00:52:10,780 CELL DIVISION HAPPENED BUT EPOB 1189 00:52:10,780 --> 00:52:14,150 STABILIZED ENOUGH SO IT DOESN'T 1190 00:52:14,150 --> 00:52:16,018 DEPOLYMERIZE COMPLETING AND CELL 1191 00:52:16,018 --> 00:52:17,186 CYCLE CAN'T MOVE TO THE NEXT 1192 00:52:17,186 --> 00:52:19,088 STAGE IS WHAT IS HAPPENING OR IS 1193 00:52:19,088 --> 00:52:19,488 THE DESIGN. 1194 00:52:19,488 --> 00:52:21,357 >> OKAY. TO STABILIZE. 1195 00:52:21,357 --> 00:52:23,859 >> YEAH, YEAH. 1196 00:52:23,859 --> 00:52:28,030 >> THANKS. 1197 00:52:28,030 --> 00:52:30,466 >> THAT WAS BEAUTIFUL. I HAD A 1198 00:52:30,466 --> 00:52:32,835 QUESTION ABOUT YOUR SPIKE 1199 00:52:32,835 --> 00:52:34,003 INDUCED MEMBRANE DEFORMATION AND 1200 00:52:34,003 --> 00:52:36,005 NOW WITH YOUR SYSTEM DO YOU HAVE 1201 00:52:36,005 --> 00:52:38,340 ANY INTENTION OF LOOKING AT 1202 00:52:38,340 --> 00:52:40,242 DIFFERENT AT LIKE FOR EXAMPLE 1203 00:52:40,242 --> 00:52:42,511 DIFFERENT RESIDUES OR SEGMENTS 1204 00:52:42,511 --> 00:52:44,413 OF SPIKE AND HOW THEY CONTRIBUTE 1205 00:52:44,413 --> 00:52:46,849 TO THE MEMBRANE DEFORMATION. 1206 00:52:46,849 --> 00:52:50,619 >> TO BE HONEST, WE HAVE MOVED 1207 00:52:50,619 --> 00:52:51,554 ON FROM SPIKE SO. 1208 00:52:51,554 --> 00:52:54,690 >> NO PROBLEM. 1209 00:52:54,690 --> 00:53:05,234 >> SORRY IT WAS A GOOD CLOSURE. 1210 00:53:06,435 --> 00:53:11,040 >> HI. I'M INTERESTED IN THE 1211 00:53:11,040 --> 00:53:12,808 SPIKE PROTEIN BINDING. YOU SAID 1212 00:53:12,808 --> 00:53:14,844 IT IS NOT REALLY CLEAR IF IT IS 1213 00:53:14,844 --> 00:53:16,545 BINDING TO THE CURVATURE OR 1214 00:53:16,545 --> 00:53:17,880 INDUCING THE CURVATURE. 1215 00:53:17,880 --> 00:53:19,882 DO YOU HAVE ANY SENSE WHICH IS 1216 00:53:19,882 --> 00:53:20,649 MORE DOMINANT? 1217 00:53:20,649 --> 00:53:24,019 >> I HONESTLY THINK IT IS 1218 00:53:24,019 --> 00:53:26,322 INDUCED INDIRECTLY INDUCING THE 1219 00:53:26,322 --> 00:53:26,622 CURVATURE. 1220 00:53:26,622 --> 00:53:29,158 SO IT WOULD BIND. IN MY MIND 1221 00:53:29,158 --> 00:53:31,894 WHAT I THINK IS IT WOULD BIND 1222 00:53:31,894 --> 00:53:33,996 STOKE AFTICALLY AND SOMETIMES IT 1223 00:53:33,996 --> 00:53:36,766 BINDS TO LOCK INTO INTEG GRIN SO 1224 00:53:36,766 --> 00:53:41,170 INTEGRIN GETS ACTIVATED AND DOT 1225 00:53:41,170 --> 00:53:42,438 INDUCES CURVATURE MAKING 1226 00:53:42,438 --> 00:53:45,808 [INDISCERNIBLE] TO BE FORMED. 1227 00:53:45,808 --> 00:53:48,444 SPIKE ITSELF DOES NOT CAUSE 1228 00:53:48,444 --> 00:53:49,945 CURVATURE FORMATION AND 1229 00:53:49,945 --> 00:53:53,082 ACTIVATING INTEG GRIN IT INDUCES 1230 00:53:53,082 --> 00:53:54,216 CURVATURE FORMATION. 1231 00:53:54,216 --> 00:53:56,252 >> MIGHT THERE BE A CERTAIN 1232 00:53:56,252 --> 00:53:58,087 AMOUNT OF FLUCTUATION NORMALLY 1233 00:53:58,087 --> 00:54:00,856 AND WHEN IT HITS A CURVE THING 1234 00:54:00,856 --> 00:54:03,058 SPIKE BINDS IT AND LOCKS IT IN 1235 00:54:03,058 --> 00:54:04,260 OR SOMETHING LIKE THAT. 1236 00:54:04,260 --> 00:54:06,362 >> YEAH INTEGRIN ENHANCES IT. 1237 00:54:06,362 --> 00:54:09,098 >> THANK YOU. 1238 00:54:09,098 --> 00:54:12,034 >> YEAH. 1239 00:54:12,034 --> 00:54:21,710 >> ANY OTHER QUESTIONS? 1240 00:54:21,710 --> 00:54:25,214 OKAY. I WOULD LIKE TO THANK DR. 1241 00:54:25,214 --> 00:54:27,349 MAZUNO FOR YOUR WONDERFUL 1242 00:54:27,349 --> 00:54:28,284 PRESENTATION AND FOR EVERYONE 1243 00:54:28,284 --> 00:54:31,587 COMING TO ATTEND TODAY. 1244 00:54:31,587 --> 00:54:32,388 >> THANK YOU. 1245 00:54:32,388 --> 00:54:42,798 >> AUDIENCE: [APPLAUSE]