1 00:00:05,560 --> 00:00:08,760 WELCOME TO THE WEDNESDAY 2 00:00:08,760 --> 00:00:10,720 AFTERNOON LECTURE SERIES. 3 00:00:10,720 --> 00:00:12,120 I'M ELLA, [INDISCERNIBLE] A 4 00:00:12,120 --> 00:00:13,720 SENIOR INVESTIGATOR LEADING THE 5 00:00:13,720 --> 00:00:17,040 SECTION ON CELLULAR 6 00:00:17,040 --> 00:00:17,560 COMMUNICATION IN NICHD. 7 00:00:17,560 --> 00:00:19,640 ON BEHALF OF THE WOMEN SCIENTIST 8 00:00:19,640 --> 00:00:22,960 ADVISORS, I AM THRILLED TO 9 00:00:22,960 --> 00:00:25,720 INTRODUCE TODAY'S WALS SPEAKER 10 00:00:25,720 --> 00:00:27,600 DR. JESSICA TREISMAN WHO HAS 11 00:00:27,600 --> 00:00:28,640 REVEALED INCREDIBLE INSIGHTS 12 00:00:28,640 --> 00:00:30,600 INTO THE CELLULAR SIGNALING FROM 13 00:00:30,600 --> 00:00:31,240 STUDIES USING THE DROSOPHILA 14 00:00:31,240 --> 00:00:32,920 MODEL SYSTEM. 15 00:00:32,920 --> 00:00:34,360 DR. TREISMAN IS A PROFESSOR IN 16 00:00:34,360 --> 00:00:35,280 THE DEPARTMENT OF CELL BIOLOGY 17 00:00:35,280 --> 00:00:38,000 AT THE NEW YORK UNIVERSITY GROON 18 00:00:38,000 --> 00:00:40,760 SSMAN SCHOOL OF MEDICINE, SHE'S 19 00:00:40,760 --> 00:00:42,720 A REVERED SCIENTIST IN THE 20 00:00:42,720 --> 00:00:44,320 NEUROAND CELLULAR BIOLOGY 21 00:00:44,320 --> 00:00:46,000 BECAUSE SHE UNCOVERED GENERAL 22 00:00:46,000 --> 00:00:47,520 FUNDAMENTAL MECHANISMS BY WHICH 23 00:00:47,520 --> 00:00:48,720 CELLS COMMUNICATE. 24 00:00:48,720 --> 00:00:50,360 USING THE VISUAL SYSTEM OF THE 25 00:00:50,360 --> 00:00:53,240 FRUIT FLY AS PRIMARY MODEL, HER 26 00:00:53,240 --> 00:00:55,480 GROUP HAS IDENTIFIED NOVEL 27 00:00:55,480 --> 00:00:58,080 COMPONENTS OF SEVERAL SIGNALING 28 00:00:58,080 --> 00:00:59,720 PATHWAYS INCLUDING HEDGE HOG, 29 00:00:59,720 --> 00:01:01,800 WIND, NOTCH, AND EPIDERMAL 30 00:01:01,800 --> 00:01:03,520 GROWTH FACTOR, SHE WORKED OUT 31 00:01:03,520 --> 00:01:06,440 THE ROLE OF THESE NEW COMPONENTS 32 00:01:06,440 --> 00:01:07,760 DURING PATTERNING AND SYNAPSE 33 00:01:07,760 --> 00:01:09,160 FORMATION AND CONNECTED THEIR 34 00:01:09,160 --> 00:01:10,880 MISREGULATION TO HUMAN DISEASES. 35 00:01:10,880 --> 00:01:13,520 HER GROUP ALSO STUDIED HOW 36 00:01:13,520 --> 00:01:15,920 VARIOUS SIGNALS ARE TRANSLATED 37 00:01:15,920 --> 00:01:19,000 INTO CELL DECISIONS FOR 38 00:01:19,000 --> 00:01:19,800 TRANSCRIPTIONAL OR POST 39 00:01:19,800 --> 00:01:20,440 TRANSCRIPTIONAL MODULATION OF 40 00:01:20,440 --> 00:01:22,680 GENE EXPRESSION. 41 00:01:22,680 --> 00:01:24,000 FROM SYNAPTIC PARTNERS SELECTION 42 00:01:24,000 --> 00:01:25,680 TO CONTRACTILE FORCES THAT 43 00:01:25,680 --> 00:01:27,520 CONTROL TISSUE MORPH O GENESIS 44 00:01:27,520 --> 00:01:29,720 TO CELL FATE DETERMINE NATION 45 00:01:29,720 --> 00:01:31,000 AND SYNAPSE ASSEMBLY, HER 46 00:01:31,000 --> 00:01:35,120 INSIGHTS ARE TRULY INSPIRING. 47 00:01:35,120 --> 00:01:37,160 DR. TREISMAN RECEIVED HER 48 00:01:37,160 --> 00:01:39,120 UNDERGRAD DEGREE FROM OXFORD 49 00:01:39,120 --> 00:01:41,680 UNIVERSITY AND HER Ph.D. FROM 50 00:01:41,680 --> 00:01:42,520 ROCKEFELLER UNIVERSITY WORKING 51 00:01:42,520 --> 00:01:43,560 WITH [INDISCERNIBLE]. 52 00:01:43,560 --> 00:01:45,840 AFTER A POST DOCTORAL FELLOWSHIP 53 00:01:45,840 --> 00:01:48,120 IN THE LABORATORY OF GENERAL 54 00:01:48,120 --> 00:01:53,280 RUBEIN AT UC BERKELEY, DR. T 55 00:01:53,280 --> 00:01:54,480 REISMAN MOVED BACK TO NEW YORK 56 00:01:54,480 --> 00:01:57,480 IN 1996 TO TAKE UP POSITION AT 57 00:01:57,480 --> 00:02:00,200 FACULTY NYU SCHOOL OF 58 00:02:00,200 --> 00:02:01,160 BIOMOLECULAR MEDICINE. 59 00:02:01,160 --> 00:02:03,800 SHE QUICKLY ROSE THROUGH THE 60 00:02:03,800 --> 00:02:05,480 RANKS AT THE INSTITUTE BECOMING 61 00:02:05,480 --> 00:02:09,240 FULL PROFESSOR THERE IN 2007 AND 62 00:02:09,240 --> 00:02:10,160 DIRECTOR IN TWEBT 20. 63 00:02:10,160 --> 00:02:13,600 IN ADDITION TO HER INSIGHTFUL 64 00:02:13,600 --> 00:02:15,480 RESEARCH, DR. TREISMAN IS A 65 00:02:15,480 --> 00:02:16,560 STELLAR SCIENTIFIC CITIZEN. 66 00:02:16,560 --> 00:02:19,040 SHE SERVED AS NUMEROUS NIH STUDY 67 00:02:19,040 --> 00:02:22,440 SECTION HAS BEEN AN ADHOC MEMBER 68 00:02:22,440 --> 00:02:24,080 OF THE VIEW COMMITTEES, HAS BEEN 69 00:02:24,080 --> 00:02:26,840 A MEMBER OF SEVERAL EDITORIAL 70 00:02:26,840 --> 00:02:29,640 BOARD AND ORGANIZED MANY 71 00:02:29,640 --> 00:02:30,240 IMPORTANT DROSOPHILA 72 00:02:30,240 --> 00:02:30,520 CONFERENCES. 73 00:02:30,520 --> 00:02:32,840 AND I WANT TO LEAVE YOU TODAY 74 00:02:32,840 --> 00:02:37,520 WITH SOMETHING I FOUND 75 00:02:37,520 --> 00:02:42,120 FASCINATING AND INSPIRING. 76 00:02:42,120 --> 00:02:44,200 JESSICA'S MOTHER [INDISCERNIBLE] 77 00:02:44,200 --> 00:02:46,000 WAS A FAMOUS PSYCHOLOGIST WHO 78 00:02:46,000 --> 00:02:46,720 SPECIALIZED IN COGNITIVE 79 00:02:46,720 --> 00:02:47,280 PSYCHOLOGY. 80 00:02:47,280 --> 00:02:50,720 SHE GAVE A WALS TALK IN 81 00:02:50,720 --> 00:02:51,400 JANUARY 1997. 82 00:02:51,400 --> 00:02:54,400 AS FAR AS I AND THE WALS OFFICE 83 00:02:54,400 --> 00:02:56,360 CAN TELL, TODAY MARKS THE FIRST 84 00:02:56,360 --> 00:02:58,800 TIME A DAUGHTER OF A PAST WALS 85 00:02:58,800 --> 00:03:02,160 SPEAKER HAS FOLLOWED IN HER 86 00:03:02,160 --> 00:03:03,040 MOTHER'S FOOT STEPS. 87 00:03:03,040 --> 00:03:05,160 JESSICA THANK YOU SO MUCH FOR 88 00:03:05,160 --> 00:03:05,600 COMING. 89 00:03:05,600 --> 00:03:11,200 WE LOOK FORWARD TO YOUR TALK. 90 00:03:11,200 --> 00:03:15,400 THANK YOU. 91 00:03:15,400 --> 00:03:16,480 [ APPLAUSE ] 92 00:03:16,480 --> 00:03:17,720 >>WELL, THANK YOU VERY MUCH FOR 93 00:03:17,720 --> 00:03:19,320 THE KIND INTRODUCTION AND I AM 94 00:03:19,320 --> 00:03:19,920 ESPECIALLY HONORED TO BE 95 00:03:19,920 --> 00:03:23,520 FOLLOWING IN MY MOTHER'S FOOT 96 00:03:23,520 --> 00:03:24,040 STEPS HERE. 97 00:03:24,040 --> 00:03:25,120 SO TODAY I'M GOING TO TRY TO 98 00:03:25,120 --> 00:03:27,840 TELL YOU ABOUT A QUITE NEW 99 00:03:27,840 --> 00:03:29,400 PROJECT IN MY LAB, IN WHICH 100 00:03:29,400 --> 00:03:32,200 WE'RE TRYING TO UNDERSTAND THE 101 00:03:32,200 --> 00:03:33,920 DEVELOPMENT OF THE DROSOPHILA 102 00:03:33,920 --> 00:03:35,800 CORNEAL LENS AND SEE IF IT CAN 103 00:03:35,800 --> 00:03:37,200 TELL US SOMETHING ABOUT THE 104 00:03:37,200 --> 00:03:38,480 HUMAN CORNEA AND USUALLY I DON'T 105 00:03:38,480 --> 00:03:40,800 HAVE TO SPEND TOO MUCH TIME 106 00:03:40,800 --> 00:03:41,520 CONVINCING A AUDIENCE OF 107 00:03:41,520 --> 00:03:43,040 SCIENTISTS THAT THE CORNEA IS 108 00:03:43,040 --> 00:03:45,560 IMPORTANT BECAUSE I BET THAT IF 109 00:03:45,560 --> 00:03:49,120 MOST OF YOU TOOK OFF YOUR 110 00:03:49,120 --> 00:03:50,520 CORRECTIVE LENSS, YOU WOULD SEE 111 00:03:50,520 --> 00:03:51,560 THIS IMAGE LOOKING SOMETHING 112 00:03:51,560 --> 00:03:54,360 LIKE THIS AND THAT'S BECAUSE THE 113 00:03:54,360 --> 00:03:55,760 CORNEA CONTRIBUTES 70% OF THE 114 00:03:55,760 --> 00:03:57,200 REFRACTIVE POWER OF THE EYE AND 115 00:03:57,200 --> 00:04:01,440 SO DEFECTS IN THE CORNEA, LEAD 116 00:04:01,440 --> 00:04:05,320 TO MANY DIFFERENT KINDS OF 117 00:04:05,320 --> 00:04:05,840 VISUAL REFRACTIVE ERRORS. 118 00:04:05,840 --> 00:04:08,800 SO IF THE CORNEA'S TOO STRONGLY 119 00:04:08,800 --> 00:04:10,280 CURVED, IT FOCUSES THE IMAGE IN 120 00:04:10,280 --> 00:04:14,880 FRONT OF THE RETINA, GIVING RISE 121 00:04:14,880 --> 00:04:16,880 TO MYOPIA, IF IT'S NOT CURVED 122 00:04:16,880 --> 00:04:18,880 ENOUGH, THE IMAGE WILL FOCUS 123 00:04:18,880 --> 00:04:23,040 BEHIND THE RETINA GIVING RISE TO 124 00:04:23,040 --> 00:04:27,400 HYPER OPILOT PROJECTA AND IF 125 00:04:27,400 --> 00:04:30,160 IT'S DISTORTED THEN THIS IS 126 00:04:30,160 --> 00:04:31,040 ASTIGMATISM. 127 00:04:31,040 --> 00:04:33,800 THE FLY CORNEAL LENS ADDED FIRST 128 00:04:33,800 --> 00:04:35,080 GLANCE MIGHT LOOK DIFFERENT FROM 129 00:04:35,080 --> 00:04:36,400 THE HUMAN CORNEA BUT THEY DO 130 00:04:36,400 --> 00:04:38,000 HAVE SOME THINGS IN COMMON AND 1 131 00:04:38,000 --> 00:04:40,560 IS THAT THEY'RE MOSTLY COMPOSED 132 00:04:40,560 --> 00:04:41,640 OF EXTRA CELLULAR MATRIX, SO IF 133 00:04:41,640 --> 00:04:44,920 YOU LOOK AT A SECTION THROUGH 134 00:04:44,920 --> 00:04:46,640 THE HUMAN CORNEA, YOU CAN SEE 135 00:04:46,640 --> 00:04:47,880 THAT ABOUT 90% OF THE THICKNESS 136 00:04:47,880 --> 00:04:51,280 IS THE STROMAL LAYER, WHICH IS 137 00:04:51,280 --> 00:04:53,800 AN EXTRA CELLULAR MATRIX MADE UP 138 00:04:53,800 --> 00:04:58,920 OF COLLAGEN FIBERS AND SMALL 139 00:04:58,920 --> 00:05:00,080 LEUCINE RICH PROTEOGLYCANS WITH 140 00:05:00,080 --> 00:05:01,480 SPORADO SIGHTS AND SEVERAL 141 00:05:01,480 --> 00:05:05,120 LAYERS ABOVE AND BELOW IT AND 142 00:05:05,120 --> 00:05:07,560 THE FLY CORNEAL LENS IS AN 143 00:05:07,560 --> 00:05:08,240 ACELLULAR, EXTRA CELLULAR 144 00:05:08,240 --> 00:05:12,400 MATRIX, IN THIS CASE, THE FIBERS 145 00:05:12,400 --> 00:05:16,240 ARE MADE UP OF KITEN, WHICH IS A 146 00:05:16,240 --> 00:05:18,000 POLYSACCHARIDE AND THEIR 147 00:05:18,000 --> 00:05:18,560 AVAILABLE CUTICLE PROTEINS. 148 00:05:18,560 --> 00:05:19,840 ABOUT YOU WE REALLY KNOW VERY 149 00:05:19,840 --> 00:05:22,360 LITTLE ABOUT THE COMPOSITION OF 150 00:05:22,360 --> 00:05:25,920 THIS CORNEAL LENS IS WHY IT 151 00:05:25,920 --> 00:05:26,800 DEVELOPED SUCH PRECISE 152 00:05:26,800 --> 00:05:28,160 CURVEATURE AND WE BEGAN TO 153 00:05:28,160 --> 00:05:29,160 TACKLE THIS AND 1 OF THE FIRST 154 00:05:29,160 --> 00:05:30,680 STEPS WE TOOK WAS TO TRY TO 155 00:05:30,680 --> 00:05:32,960 UNDERSTAND HOW THE CELLS THAT 156 00:05:32,960 --> 00:05:36,560 SECRETE THIS CORNEAL LENS 157 00:05:36,560 --> 00:05:40,280 DEVELOP ISSUES SO THESE WOULD BE 158 00:05:40,280 --> 00:05:41,840 THE UNDERLYING CO-CELLS AND 159 00:05:41,840 --> 00:05:45,120 PIGMENT CELLS. 160 00:05:45,120 --> 00:05:47,440 SO, THIS IS WHETHER THERE'S A 161 00:05:47,440 --> 00:05:48,720 SIGNIFICANT DIFFERENCE BETWEEN 162 00:05:48,720 --> 00:05:53,120 THE DROSOPHILA AND MAMMALIAN 163 00:05:53,120 --> 00:05:53,360 SYSTEMS. 164 00:05:53,360 --> 00:05:55,680 THE MAMMALIAN DEVELOPED FROM THE 165 00:05:55,680 --> 00:05:57,280 EPITHELIUM AFTER THE LENS IS 166 00:05:57,280 --> 00:05:58,800 INVAGINATED AND IT'S QUITE 167 00:05:58,800 --> 00:06:00,960 SEPARATE FROM THE RETINA WHICH 168 00:06:00,960 --> 00:06:03,040 DEVELOPS FROM THE NEURAL TUBE. 169 00:06:03,040 --> 00:06:08,400 BUT IN FLIES, A SINGLE 170 00:06:08,400 --> 00:06:09,240 EPITHELIUM DISK 171 00:06:09,240 --> 00:06:11,120 THE DISK GIVES RISE FIRST TO 172 00:06:11,120 --> 00:06:15,920 THE 8 PHOTO RECEPTORS IN EACH 173 00:06:15,920 --> 00:06:17,480 UNIT OF THE COMPOUND EYE SO 174 00:06:17,480 --> 00:06:19,200 THESE ARE ALREADY, 2, 5, 3, AND 175 00:06:19,200 --> 00:06:22,240 4, AND 1 AND 6 AND 7. 176 00:06:22,240 --> 00:06:23,520 AND THEN THE SAME EPITHELIUM 177 00:06:23,520 --> 00:06:26,400 WILL GIVE PRIZE TO THE 4 CONE 178 00:06:26,400 --> 00:06:28,000 CELLS AND 2 PRIMARY PIGMENT 179 00:06:28,000 --> 00:06:29,160 CELLS AND A LAT ISOTOPE OF 180 00:06:29,160 --> 00:06:30,640 SECONDARY AND TERTIARY CELLS 181 00:06:30,640 --> 00:06:34,520 WHICH WILL COMPLETE THE CORNEAL 182 00:06:34,520 --> 00:06:35,320 LENS. 183 00:06:35,320 --> 00:06:37,160 AND SO, THIS HAPPENS AS A WAVE 184 00:06:37,160 --> 00:06:42,560 OF DEVELOPMENT PROGRESSES ACROSS 185 00:06:42,560 --> 00:06:45,400 THE IIMAGINAL DISK GIVING RISE 186 00:06:45,400 --> 00:06:46,680 TO CELL RISE AND SUCCESS, AND 187 00:06:46,680 --> 00:06:48,680 THAT MEANS THAT A 188 00:06:48,680 --> 00:06:49,760 UNDIFFERENTIATED PROGENITOR IN 189 00:06:49,760 --> 00:06:51,080 THE INTERIOR OF THIS EYE DISK 190 00:06:51,080 --> 00:06:52,680 CAN GIVE RISE TO A NUMBER OF 191 00:06:52,680 --> 00:06:54,320 DIFFERENT CELL FATES AND THIS IS 192 00:06:54,320 --> 00:06:56,400 A QUESTION MY FORMER STUDENT 193 00:06:56,400 --> 00:06:57,920 CAROLYN MORRISON WAS INTERESTED 194 00:06:57,920 --> 00:07:00,200 IN UNDERSTANDING, HOW IS IT THE 195 00:07:00,200 --> 00:07:01,960 SINGLE PROGENITOR COULD GIVE 196 00:07:01,960 --> 00:07:03,080 RISE TO EITHER PHOTO RECEPTOR 197 00:07:03,080 --> 00:07:06,200 WHICH IS A LIGHT SENSITIVE 198 00:07:06,200 --> 00:07:09,040 NEURON, OR CONE CELL, WHICH IS A 199 00:07:09,040 --> 00:07:12,040 GLIAL CELL THAT CAN SECRETE 200 00:07:12,040 --> 00:07:13,800 CORNEAL LENS COMPONENTS OR A 201 00:07:13,800 --> 00:07:16,200 PIGMENT CELL WHICH WILL PRODUCE 202 00:07:16,200 --> 00:07:20,520 SCREENING PIGMENTS AND RECYCLE 203 00:07:20,520 --> 00:07:21,680 RETINAL INDIVIDUAL CYCLE. 204 00:07:21,680 --> 00:07:22,880 SO CAROLYN HAD 1 CLUE TO START 205 00:07:22,880 --> 00:07:25,200 WITH, WHICH WAS A PROTEIN THAT 206 00:07:25,200 --> 00:07:27,680 WAS DISCOVERED BACK IN 1989 BY 207 00:07:27,680 --> 00:07:29,880 DEVIN MOSES AND THE RUBEIN LAB 208 00:07:29,880 --> 00:07:32,600 AND THIS IS A PROTEIN CALLED 209 00:07:32,600 --> 00:07:33,920 GLASS, IT'S A TRANSCRIPTION 210 00:07:33,920 --> 00:07:35,880 FACTOR AND YOU CAN SEE THAT IN A 211 00:07:35,880 --> 00:07:37,760 GLASS MUTANT THE EYE IS MUCH 212 00:07:37,760 --> 00:07:41,840 SMALLER AND MORE DISORDERED THAN 213 00:07:41,840 --> 00:07:42,480 A WILD-TYPE EYE. 214 00:07:42,480 --> 00:07:45,360 AND IF YOU SECTION THROUGH AN 215 00:07:45,360 --> 00:07:48,080 EYE THAT'S A MOSAIC OF BOTH 216 00:07:48,080 --> 00:07:49,240 WILD-TYPE AND GLASS MUTANT 217 00:07:49,240 --> 00:07:51,400 TISSUE, YOU CAN SEE THAT IN THE 218 00:07:51,400 --> 00:07:55,120 WILD-TYPE REGION, YOU SEE THE 219 00:07:55,120 --> 00:07:57,200 PHOTO RECEPTOR RAB DO MERE SO 220 00:07:57,200 --> 00:08:01,880 THESE BLOCKS ARE THE LIGHT 221 00:08:01,880 --> 00:08:03,120 DETECTING ORGANELLE THAT'S 222 00:08:03,120 --> 00:08:04,320 OBVIOUS TO A WATER CONE OUT OF 223 00:08:04,320 --> 00:08:06,600 SEGMENT BUT IN THE REGION, THESE 224 00:08:06,600 --> 00:08:09,480 RAB DO MERES ARE COMPLETELY 225 00:08:09,480 --> 00:08:14,080 MISSING AND IT'S THOUGHT THAT 226 00:08:14,080 --> 00:08:15,280 THE GLASS RECTIFYS THE CELL 227 00:08:15,280 --> 00:08:15,640 PHOTO MATE. 228 00:08:15,640 --> 00:08:18,400 AND IF YOU LOOK AT THE PHOTO 229 00:08:18,400 --> 00:08:20,040 RECEPTOR GENES THEY SEEM TO BE 230 00:08:20,040 --> 00:08:22,200 COMPLETELY MISSING IN THE 231 00:08:22,200 --> 00:08:22,480 MUTANTS. 232 00:08:22,480 --> 00:08:24,880 SO FOR EXAMPLE, K-ROSETTA AND IN 233 00:08:24,880 --> 00:08:27,360 THE IMAGINAL DISK MISSING IN 234 00:08:27,360 --> 00:08:29,800 GLASS MUTANTS, ROUGH ATOM DOBSIN 235 00:08:29,800 --> 00:08:31,840 1 IN THE ADULT RETINA IS ALSO 236 00:08:31,840 --> 00:08:34,520 COMPLETELY GONE. 237 00:08:34,520 --> 00:08:36,280 SO, THERE'S A COUPLE OF GENES 238 00:08:36,280 --> 00:08:39,360 BUT WE WANT TO GET THEM ALL 239 00:08:39,360 --> 00:08:40,480 COMPLETE PICTURE OF WHAT THIS 240 00:08:40,480 --> 00:08:42,960 WAS COLLECTING AND SO WE 241 00:08:42,960 --> 00:08:46,960 COLLABORATED WITH MARTIN AT 242 00:08:46,960 --> 00:08:48,760 BAYLOR AND KUMA R IN THEIR LAB 243 00:08:48,760 --> 00:08:52,200 AND THEY DID A SINGLE CELL SEQ 244 00:08:52,200 --> 00:08:54,360 ANALYSIS OF WILD-TYPE AND SINGLE 245 00:08:54,360 --> 00:08:55,720 GLASS MUTANT DISKS AND THIS WAS 246 00:08:55,720 --> 00:08:56,680 AT THE END LEVEL STAGE. 247 00:08:56,680 --> 00:08:58,440 SO IN THE WILD-TYPE EYE DISKS 248 00:08:58,440 --> 00:09:00,600 THEY COULD SEE SEPARATE CLUSTERS 249 00:09:00,600 --> 00:09:04,040 REPRESENTING EACH OF THE PHOTO 250 00:09:04,040 --> 00:09:06,240 RECEPTOR CELL TYPES, SO R8, 2 251 00:09:06,240 --> 00:09:08,240 AND 5, 3 AND 4 AND 1 AND 6 AND 7 252 00:09:08,240 --> 00:09:11,120 AS WELL AS THE CURRENT CELLS, 253 00:09:11,120 --> 00:09:12,880 AND EACH OF THESE CLUSTER SYSTEM 254 00:09:12,880 --> 00:09:14,200 ACTUALLY A TRAJECTORY FROM LESS 255 00:09:14,200 --> 00:09:15,840 DIFFERENTIATED ON THE LEFT TO 256 00:09:15,840 --> 00:09:22,200 MORE DIFFERENTIATED ON THE 257 00:09:22,200 --> 00:09:22,400 RIGHT. 258 00:09:22,400 --> 00:09:25,240 IN GLASS MUTANT DISKSOT OTHER 259 00:09:25,240 --> 00:09:26,800 HAND THE ONLY RAJECTORY FOR THE 260 00:09:26,800 --> 00:09:28,680 OTHER WAS THE R8 FOR TRAJECTORY 261 00:09:28,680 --> 00:09:33,080 AND A SMALLER CONE TRAJECTORY. 262 00:09:33,080 --> 00:09:35,320 SO THERE ARE EARLY MARKERS OF 263 00:09:35,320 --> 00:09:37,360 R2, 3, AND 4, FOR THE PHOTO 264 00:09:37,360 --> 00:09:40,000 RECEPTORS BUT NO EXTENDED 265 00:09:40,000 --> 00:09:40,880 TRAJECTORY OF DIFFERENTIATION 266 00:09:40,880 --> 00:09:43,520 AND A 1, 6, AND 7 SEEM TO BE 267 00:09:43,520 --> 00:09:48,080 COLONEL PLETELY MISSING. 268 00:09:48,080 --> 00:09:49,160 --COMPLETELY MISSING. 269 00:09:49,160 --> 00:09:54,080 SO I WILL SHOW A FEW GENES THAT 270 00:09:54,080 --> 00:09:54,400 IS BASED ON. 271 00:09:54,400 --> 00:09:55,840 SO [INDISCERNIBLE] IS A 272 00:09:55,840 --> 00:09:57,680 PARTICULAR GENE, AND WE KNOW 273 00:09:57,680 --> 00:09:59,760 THAT REPRESENTS THE R8 IN 274 00:09:59,760 --> 00:10:00,920 WILD-TYPE AND GLASS MUTANT AND 275 00:10:00,920 --> 00:10:03,240 IF WE LOOK AT EARLY PHOTO 276 00:10:03,240 --> 00:10:04,440 RECEPTOR SPECIFIC GENES SUCH AS 277 00:10:04,440 --> 00:10:06,160 SCRATCH, YOU CAN SEE THAT THIS 278 00:10:06,160 --> 00:10:09,320 IS COMING ON IN ALL THE PHOTO 279 00:10:09,320 --> 00:10:10,680 RECEPTOR TRAJECTORIES AND 280 00:10:10,680 --> 00:10:12,680 WILD-TYPE AND IN SCRATCH, WE SEE 281 00:10:12,680 --> 00:10:14,680 IT IN R8 AND IT ALSO TURNS ON IN 282 00:10:14,680 --> 00:10:16,480 THIS REGION WHERE OUR R23, 5, 283 00:10:16,480 --> 00:10:20,360 AND 4 ARE JUST BEGINNING TO 284 00:10:20,360 --> 00:10:20,720 DIFFERENTIATE. 285 00:10:20,720 --> 00:10:22,280 BUT LATER PHOTO RECEPTOR 286 00:10:22,280 --> 00:10:26,840 SPECIFIC GENES SUCH AS RETAINED 287 00:10:26,840 --> 00:10:29,080 IN THE GLASS MUTANT ARE ONLY 288 00:10:29,080 --> 00:10:29,680 PRESENT IN R8. 289 00:10:29,680 --> 00:10:30,880 SO GLASS SEEMS TO BE NECESSARY 290 00:10:30,880 --> 00:10:32,760 FOR THE DIFREBTIATION OF ALL THE 291 00:10:32,760 --> 00:10:38,200 PHOTO RECEPTORS OTHER THAN R8. 292 00:10:38,200 --> 00:10:40,040 SO THAT'S WHAT GLASS IS 293 00:10:40,040 --> 00:10:40,400 NECESSARY FOR. 294 00:10:40,400 --> 00:10:41,440 CAROLYN WANTED TO SEE WHAT IT 295 00:10:41,440 --> 00:10:43,720 WAS SUFFICIENT TO DO. 296 00:10:43,720 --> 00:10:46,760 AND SO SHE MISS EXPRESSED GLASS 297 00:10:46,760 --> 00:10:49,920 IN A DIFFERENT TISSUE, THE WING 298 00:10:49,920 --> 00:10:52,600 DISK, TO SEE WHAT IT COULD 299 00:10:52,600 --> 00:10:57,640 INDUCE IN THE ECTOPIC SITUATION. 300 00:10:57,640 --> 00:11:00,280 SO HERE THE GREEN CELLS 301 00:11:00,280 --> 00:11:03,520 EXPRESSING BOTH GLASS AND GFP AS 302 00:11:03,520 --> 00:11:05,520 AN IMAGINAL DISK AND SHE SAW 303 00:11:05,520 --> 00:11:06,640 THAT KOPTICAL IMAGESIN, THAT THE 304 00:11:06,640 --> 00:11:10,400 SPECIFIC GENE WAS TURNED ON BY 305 00:11:10,400 --> 00:11:12,080 GLASS. 306 00:11:12,080 --> 00:11:16,440 IN A WILD-TYPE EYE DISK, KOPTIN 307 00:11:16,440 --> 00:11:18,440 WOULD COME ON IF THE CELLS HAD 308 00:11:18,440 --> 00:11:21,640 ALREADY BEEN EXPRESSING NEURONAL 309 00:11:21,640 --> 00:11:24,160 MARKERS SUCH AS ELAV FOR QUITE A 310 00:11:24,160 --> 00:11:26,960 WHILE BUT IN THE WING DISK ELAV 311 00:11:26,960 --> 00:11:28,360 WAS NOT INDUCED BY A TOPIC 312 00:11:28,360 --> 00:11:30,760 GLASS, SO THIS CAN INDUCE THE 313 00:11:30,760 --> 00:11:32,600 PHOTO RECEPTOR GENES BUT IT'S 314 00:11:32,600 --> 00:11:33,800 NOT TRANSFORMING THE CELLS INTO 315 00:11:33,800 --> 00:11:35,200 PHOTO RECEPTORS BECAUSE THEY'RE 316 00:11:35,200 --> 00:11:38,800 NOT TAKING ON A NEURONAL 317 00:11:38,800 --> 00:11:39,080 IDENTITY. 318 00:11:39,080 --> 00:11:40,800 AND THIS ISN'T COMPLETELY 319 00:11:40,800 --> 00:11:41,760 SURPRISING BECAUSE GLASS 320 00:11:41,760 --> 00:11:42,960 EXPRESSION IN THE EYE DISK IS 321 00:11:42,960 --> 00:11:46,840 NOT RESTRICTED TO THE FET O 322 00:11:46,840 --> 00:11:47,240 RECEPTOR NEURONS. 323 00:11:47,240 --> 00:11:48,960 IT'S ACTUALLY EXPRESSED IN ALL 324 00:11:48,960 --> 00:11:51,840 THE CELLS OF THE POSTERIOR 325 00:11:51,840 --> 00:11:52,440 DIFFERENTIATING REGION OF THE 326 00:11:52,440 --> 00:11:53,920 DISK AS YOU CAN SEE HERE. 327 00:11:53,920 --> 00:11:56,720 SO GLASS CAN'T BE SUFFICIENT TO 328 00:11:56,720 --> 00:11:58,360 INDUCE THE PHOTO RECEPTOR NEURON 329 00:11:58,360 --> 00:11:59,200 IDENTITY, THERE MUST BE 330 00:11:59,200 --> 00:12:05,000 SOMETHING ELSE WORKING TOGETHER 331 00:12:05,000 --> 00:12:05,680 WITH IT. 332 00:12:05,680 --> 00:12:07,080 AND WE HAD A GOOD CANDIDATE FOR 333 00:12:07,080 --> 00:12:09,640 THAT WHICH IS THE GROWTH 334 00:12:09,640 --> 00:12:11,240 RECEPTOR SIGNALING PATHWAY WHICH 335 00:12:11,240 --> 00:12:12,800 MATTHEW FREEMAN AND MATTHEW 336 00:12:12,800 --> 00:12:14,880 ROSES AND OTHERS HAD SHOWED 337 00:12:14,880 --> 00:12:17,760 PHOTO RECEPTORS OTHER THAN R8 TO 338 00:12:17,760 --> 00:12:18,240 DIFFERENTIATE. 339 00:12:18,240 --> 00:12:20,400 SO AS THIS DIFFERENTIATION WAVE 340 00:12:20,400 --> 00:12:25,160 MOVES ACROSS THE EYE DISK, TURNS 341 00:12:25,160 --> 00:12:27,360 ON A TONAL WHICH IS PRONEURAL 342 00:12:27,360 --> 00:12:29,720 FACTOR AND THAT SPECIFIES THE R8 343 00:12:29,720 --> 00:12:33,680 PHOTO RECEPTOR AND R8 THEN 344 00:12:33,680 --> 00:12:36,360 PRODUCES SWITS WHICH IS EGF 345 00:12:36,360 --> 00:12:41,600 RECEPTOR AND ITS EGF RECEPTOR 346 00:12:41,600 --> 00:12:43,040 SIGNALING SO IF YOU'RE NOT GOING 347 00:12:43,040 --> 00:12:45,760 TO KNOCK OUT EGFR SIGNALING, YOU 348 00:12:45,760 --> 00:12:47,760 LOSE THE PHOTO RECEPTORS OTHER 349 00:12:47,760 --> 00:12:48,320 THAN R8. 350 00:12:48,320 --> 00:12:50,200 SIMILAR TO THE GLASS MUTANT 351 00:12:50,200 --> 00:12:50,640 PHENOTYPE. 352 00:12:50,640 --> 00:12:52,600 SO THIS IS JUST A SIMPLIFIED 353 00:12:52,600 --> 00:12:54,520 VERSION OF THE EGF RECEPTOR 354 00:12:54,520 --> 00:12:55,480 SIGNALING PATHWAY AND I JUST 355 00:12:55,480 --> 00:13:00,280 WANT TO DRAW YOUR ATTENTION TO 356 00:13:00,280 --> 00:13:00,920 TRANSCRIPTION DEFACTOR DOWN 357 00:13:00,920 --> 00:13:03,880 STREAM OF THE PATHWAY WHICH IS 358 00:13:03,880 --> 00:13:05,920 CALLED POINTED AND AN EARLY STEP 359 00:13:05,920 --> 00:13:10,360 IN THE PATHWAY, AT THE GTP 360 00:13:10,360 --> 00:13:12,240 BINDING REZ WHICH WE CAN USE TO 361 00:13:12,240 --> 00:13:13,640 ACTIVATE THE PATHWAY BECAUSE 362 00:13:13,640 --> 00:13:15,840 THERE'S A ACTIVE FORM OF IT. 363 00:13:15,840 --> 00:13:19,920 SO, WE LOOKEDDA THE WHETHER THIS 364 00:13:19,920 --> 00:13:24,680 PATHWAY COULD SYNERGIZE WITH 365 00:13:24,680 --> 00:13:25,720 GLASS TO INDUCE NEURONAL 366 00:13:25,720 --> 00:13:27,120 DIFFERENTIATION IN THE WING DISK 367 00:13:27,120 --> 00:13:28,880 AGAIN, SO GLASS ALONE DOES NOT 368 00:13:28,880 --> 00:13:33,680 TURN ON THE NEURONAL MARKER 369 00:13:33,680 --> 00:13:35,920 ELAV, IT TURNS ON ELAV IN A FEW 370 00:13:35,920 --> 00:13:38,800 CELLS BUT NOT SIGNIFICANTLY BUT 371 00:13:38,800 --> 00:13:44,920 IF WE EXPRESS BOTH TOGETHER NOW 372 00:13:44,920 --> 00:13:47,960 A LOT OF CELLS TAKE ON THE 373 00:13:47,960 --> 00:13:48,480 IDENTITY AND EXPRESS ELAV. 374 00:13:48,480 --> 00:13:50,600 AND WE CAN SHOW THIS IS 375 00:13:50,600 --> 00:13:52,000 DEPENDENT ON THE TRANSCRIPTION 376 00:13:52,000 --> 00:13:53,640 FACTOR POINTED BECAUSE IF WE 377 00:13:53,640 --> 00:13:55,560 MISS EXPRESS GLASS AND ACTIVATED 378 00:13:55,560 --> 00:13:57,640 REZ IN CELLS THAT ARE MUTANT FOR 379 00:13:57,640 --> 00:14:02,640 POINTED WE LOSE MOST OF THAT 380 00:14:02,640 --> 00:14:03,120 ECTOPIC ELAV EXPRESSION. 381 00:14:03,120 --> 00:14:05,080 SO THERE SEEMS TO BE SYNERGY 382 00:14:05,080 --> 00:14:08,480 BETWEEN IT IS 2 TRANSCRIPTION 383 00:14:08,480 --> 00:14:11,040 FACTORS GLASS AND POINTED. 384 00:14:11,040 --> 00:14:13,200 SO ELAV IS JUST 1 GENE SO MAYBE 385 00:14:13,200 --> 00:14:15,720 I SHOULDN'T MAKE SUCH A BROAD 386 00:14:15,720 --> 00:14:17,560 CONCLUSION BASED ON THAT BUT MY 387 00:14:17,560 --> 00:14:19,880 STUDENT WANTED TO KNOW WHO ELSE 388 00:14:19,880 --> 00:14:22,040 GLASS AND ACTIVATED RAS COULD 389 00:14:22,040 --> 00:14:25,840 INDUCE AND SO SHE DID RNA SEQ ON 390 00:14:25,840 --> 00:14:26,920 THOSE WING DISKS I JUST SHOWED 391 00:14:26,920 --> 00:14:29,920 YOU TO LOOK AT WHAT GENES WHEN 392 00:14:29,920 --> 00:14:31,880 DUCED BY THE SYNERGISTIC 393 00:14:31,880 --> 00:14:34,760 INTERACTION OF GLASS AND RAS, 394 00:14:34,760 --> 00:14:40,400 AND SHE FOUND 256 GENES THAT 395 00:14:40,400 --> 00:14:41,720 WERE SYNERGISTICALLY INDUCED BY 396 00:14:41,720 --> 00:14:45,040 RAS, AND THAT WERE MORE LONE AND 397 00:14:45,040 --> 00:14:47,160 WERE POINTED DEPENDENT AND AT 398 00:14:47,160 --> 00:14:49,320 LEAST 40% OF THOSE GENES HAVE NO 399 00:14:49,320 --> 00:14:50,800 NEURONAL FUNCTIONS SO THIS IS 400 00:14:50,800 --> 00:14:55,560 JUST A FEW EXAMPLES, NUEVERIN 1 401 00:14:55,560 --> 00:14:58,560 AND SCRATCHED AND NEURON 402 00:14:58,560 --> 00:15:00,200 SPECIFIC TRANSCRIPT FACTORS AND 403 00:15:00,200 --> 00:15:03,920 TAG MIN 4 AND DRIVEN, AND 404 00:15:03,920 --> 00:15:04,560 SYNAPTIC VESICLE PROTEINS, A 405 00:15:04,560 --> 00:15:06,800 TRIP IS A CHANNEL, IT'S 406 00:15:06,800 --> 00:15:08,880 IMPORTANT FOR PHOTO 407 00:15:08,880 --> 00:15:11,640 TRANSDUCTION, AND FOR THE 408 00:15:11,640 --> 00:15:12,840 MICROTUBIAL PROTEIN SPECIFIC TO 409 00:15:12,840 --> 00:15:14,480 NEURONS, SO ALL OF THESE ARE 410 00:15:14,480 --> 00:15:16,240 INDUCED MUCH MORE STRONGLY BY 411 00:15:16,240 --> 00:15:18,520 THE COMBINATION OF GLASS AND 412 00:15:18,520 --> 00:15:21,160 ACTIVATED RAS THAN BY OTHER 413 00:15:21,160 --> 00:15:21,480 ALONE. 414 00:15:21,480 --> 00:15:23,200 AND THIS EXPRESSION IS MUCH 415 00:15:23,200 --> 00:15:29,000 REDUCED IN POINTED MUTANT CELLS. 416 00:15:29,000 --> 00:15:31,600 SO NEXT, SUH WANTED TO KNOW HOW 417 00:15:31,600 --> 00:15:32,720 THIS WAS HAPPENING? 418 00:15:32,720 --> 00:15:34,320 IN A MECHANISTIC WAY AND THERE 419 00:15:34,320 --> 00:15:36,960 ARE 2 SORT OF EXTREME 420 00:15:36,960 --> 00:15:38,320 POSSIBILITIES FOR THAT, SO 421 00:15:38,320 --> 00:15:40,320 EITHER THEY COULD BE A DIRECT 422 00:15:40,320 --> 00:15:41,840 SYNERGY, WHICH BOTH GLASS AND 423 00:15:41,840 --> 00:15:42,880 POINTED WOULD BE DIRECTLY 424 00:15:42,880 --> 00:15:45,360 BINDING TO THE PROMOTERS OF ALL 425 00:15:45,360 --> 00:15:49,200 OF THESE DOWN STREAM GENES. 426 00:15:49,200 --> 00:15:50,240 OR ALTERNATIVELY, EITHER GLASS 427 00:15:50,240 --> 00:15:52,560 OR POINTED OR BOTH COULD ACT 428 00:15:52,560 --> 00:15:54,840 PRIMARILY BY INDUCING OTHER 429 00:15:54,840 --> 00:15:57,360 TRANSCRIPTION FACTORS AND THOSE 430 00:15:57,360 --> 00:15:58,840 TRANSCRIPTION FACTORS WOULD THEN 431 00:15:58,840 --> 00:16:02,720 ACT DIRECTLY ON THESE DOWN 432 00:16:02,720 --> 00:16:03,760 STREAM TARGET GENES. 433 00:16:03,760 --> 00:16:05,280 SO TO ADDRESS THIS QUESTION, SHE 434 00:16:05,280 --> 00:16:08,080 USED A TECHNIQUE CALLED TARGETED 435 00:16:08,080 --> 00:16:13,960 DEM ID, THIS IS A WAY TO LOOK AT 436 00:16:13,960 --> 00:16:14,560 TRANSCRIPTION FACTOR BINDING 437 00:16:14,560 --> 00:16:17,680 SETS IN A WAY, SO THE WHYED IS 438 00:16:17,680 --> 00:16:19,120 THAT YOU FUSE THE FACTOR THAT 439 00:16:19,120 --> 00:16:25,280 YOU'RE INTERESTED IN TO THE DAN 440 00:16:25,280 --> 00:16:28,240 E.COLI AND THAT CAN THEN 441 00:16:28,240 --> 00:16:29,200 METHALATE WHERE THE PROTEIN IS 442 00:16:29,200 --> 00:16:30,720 BOUND AND YOU CAN EXPRESS THIS 443 00:16:30,720 --> 00:16:32,000 IN SPECIFIC CELL TYPESSA THE A 444 00:16:32,000 --> 00:16:34,160 VERY LOW LEVEL SO IT DOESN'T 445 00:16:34,160 --> 00:16:37,920 DISRUPT A CELL FATE OR ANYTHING. 446 00:16:37,920 --> 00:16:40,440 AND YOU CAN USE METHYLATION TO 447 00:16:40,440 --> 00:16:46,240 THEN ISOLATE AND SEQUENCE THAT 448 00:16:46,240 --> 00:16:46,440 DNA. 449 00:16:46,440 --> 00:16:49,240 SO SUH DID THIS WITH BOTH GLASS 450 00:16:49,240 --> 00:16:51,160 AND POINTED SO SHE FUSED EACH OF 451 00:16:51,160 --> 00:16:54,920 THOSE TO THE METHYLASE AS WELL 452 00:16:54,920 --> 00:16:57,520 AS EXPRESSING DAM METHYLASE OPEN 453 00:16:57,520 --> 00:16:59,920 CONTROL AND HE DID THIS IN 3 454 00:16:59,920 --> 00:17:02,560 DIFFERENT CELL TYPES, SO 455 00:17:02,560 --> 00:17:04,120 INTERNAL IS EXPRESSED IN THE 456 00:17:04,120 --> 00:17:09,320 FIRST CELLS TO DIFFERENTIATE IN 457 00:17:09,320 --> 00:17:10,800 THE EYE DISK. 458 00:17:10,800 --> 00:17:13,560 LV4S IN THE PHOTO RECEPTORS AND 459 00:17:13,560 --> 00:17:16,000 THE SPARK IN THE CONE CELLS, AND 460 00:17:16,000 --> 00:17:18,200 TO SEE WHAT GENES CAN FIND TO IN 461 00:17:18,200 --> 00:17:20,640 ALL OF THESE DIFFERENT CELL 462 00:17:20,640 --> 00:17:21,320 TYPES. 463 00:17:21,320 --> 00:17:22,880 SO WE JUST TAKE OUT THESE DATA A 464 00:17:22,880 --> 00:17:24,240 COUPLE WEEKS -ING AND SO I CAN'T 465 00:17:24,240 --> 00:17:26,240 REALLY TELL YOU THE FULL 466 00:17:26,240 --> 00:17:27,480 ANALYSIS YET, BUT I JUST WANTED 467 00:17:27,480 --> 00:17:32,680 TO SHOW A FEW PRELIMINARY 468 00:17:32,680 --> 00:17:33,960 CONCLUSIONS SO FIRST WE CAN SAY 469 00:17:33,960 --> 00:17:35,840 THAT GLASS AND POINTED HAVE BOTH 470 00:17:35,840 --> 00:17:44,560 SHARED AND DISTINCT TARGETS SO 471 00:17:44,560 --> 00:17:47,400 FOR INSTANCE BOTH GLASS AND CONE 472 00:17:47,400 --> 00:17:49,600 BIND TO THE LIGAND GENE, AND 473 00:17:49,600 --> 00:17:52,440 THEY CONTAIN AND SHOW NECESSARY 474 00:17:52,440 --> 00:17:53,200 THE SUFFICIENT ENHANCEMENT TO 475 00:17:53,200 --> 00:18:00,560 THE EYE DISK, SO THAT WAS NICE. 476 00:18:00,560 --> 00:18:04,160 SOME GENES LIKE [INDISCERNIBLE] 477 00:18:04,160 --> 00:18:04,760 POINTED BY NOT GLASS. 478 00:18:04,760 --> 00:18:06,880 ALSO IF YOU LOOK AT THE 479 00:18:06,880 --> 00:18:08,640 DIFFERENT CELL TYPES, THERE ARE 480 00:18:08,640 --> 00:18:10,240 SOME GENES THAT ARE REGULATED OR 481 00:18:10,240 --> 00:18:12,560 BOUND BY GLASS IN ALL CELL 482 00:18:12,560 --> 00:18:17,240 TYPES, THE EARLY DIFIENTERATING 483 00:18:17,240 --> 00:18:18,760 CELLS, THE DIFFERENTIATORS AND 484 00:18:18,760 --> 00:18:20,360 THE CONE CELLS, THOSE ARE ONLY 485 00:18:20,360 --> 00:18:22,280 IN THE EARLY CELLS OR ONLY IN 486 00:18:22,280 --> 00:18:25,200 THE NEURONS OR ONLY IN THE CONE 487 00:18:25,200 --> 00:18:25,440 CELLS. 488 00:18:25,440 --> 00:18:26,600 AND SIMILARLY FOR POINTED THERE 489 00:18:26,600 --> 00:18:28,480 ARE SOME GENES THAT ARE BOUND BY 490 00:18:28,480 --> 00:18:31,080 POINTED IN ALL 3 CELL TYPES OR 491 00:18:31,080 --> 00:18:37,560 ONLY IN EARLY CELLS, NEURONS OR 492 00:18:37,560 --> 00:18:38,320 CONE CELLS. 493 00:18:38,320 --> 00:18:41,640 DSM AND TO COME BACK TO THOSE 494 00:18:41,640 --> 00:18:42,280 SYNERGISTICALLY INDUCED GENES, 495 00:18:42,280 --> 00:18:45,200 THERE ARE SOME THAT SEEM TO BE 496 00:18:45,200 --> 00:18:46,520 DIRECTLY REGULATED BY GLASS AND 497 00:18:46,520 --> 00:18:49,600 POINTED BINDING TO THE SAME SITE 498 00:18:49,600 --> 00:18:50,600 SUCH AS [INDISCERNIBLE]. 499 00:18:50,600 --> 00:18:52,480 YOU CAN SEE BINDING BY BOTH 500 00:18:52,480 --> 00:18:55,320 GLASS AND POINTED, AND THERE ARE 501 00:18:55,320 --> 00:18:57,640 OTHERS THAT APPEAR TO BE 502 00:18:57,640 --> 00:18:58,520 INDIRECTLY REGULATE SAID BECAUSE 503 00:18:58,520 --> 00:19:00,640 WE DON'T SEE BINDING OF EITHER 504 00:19:00,640 --> 00:19:02,160 GLASS OR POINTED SO THERE MUST 505 00:19:02,160 --> 00:19:04,160 BE A SECONDARY TRANSCRIPTION 506 00:19:04,160 --> 00:19:04,960 FACTOR THAT'S MEDIATING THE 507 00:19:04,960 --> 00:19:08,640 EFFECT OF GLASS AND ACTIVATED 508 00:19:08,640 --> 00:19:19,160 RAS ON EXPRESSION OF NERFIN 1. 509 00:19:20,880 --> 00:19:23,200 SO THESE ARE ALSO EXPRESSED BY 510 00:19:23,200 --> 00:19:24,440 OTHER CELL TYPES IN THE DISKS 511 00:19:24,440 --> 00:19:25,560 AND WE WONDER WHETHER IT MIGHT 512 00:19:25,560 --> 00:19:26,800 HAVE A FUNCTION AS OTHER CELL 513 00:19:26,800 --> 00:19:29,040 TYPES AS WELL AND WE COULD SHOW 514 00:19:29,040 --> 00:19:30,400 THAT IT DOES SEEM TO SO IF WE 515 00:19:30,400 --> 00:19:33,440 LOOK AT THE GENES THAT ARE 516 00:19:33,440 --> 00:19:35,920 INDUCED BY GLASS WHEN THIS TAKES 517 00:19:35,920 --> 00:19:37,880 PLACE IN THE WING DISK, WE CAN 518 00:19:37,880 --> 00:19:40,240 FIND GENES THAT ARE SPECIFIC FOR 519 00:19:40,240 --> 00:19:41,920 CONE CELLS OR PRIMARY PIGMENT 520 00:19:41,920 --> 00:19:43,280 CELLS OR SECONDARY AND TERTIARY 521 00:19:43,280 --> 00:19:45,240 PIGMENT CELLS AND THESE ARE ALL 522 00:19:45,240 --> 00:19:49,400 INDUCED BY GLASS INDEPENDENTLY 523 00:19:49,400 --> 00:19:51,480 FOR RAS ACTIVATION, SO RAS 524 00:19:51,480 --> 00:19:53,760 ACTIVATION DOES NOT FURTHER 525 00:19:53,760 --> 00:19:55,000 INDUCE THEIR EXPRESSION. 526 00:19:55,000 --> 00:19:58,240 AND WE ALSO NEW THAT GLASS HAS 527 00:19:58,240 --> 00:20:00,040 BIOLOGICAL EFFECTS ON THESE 528 00:20:00,040 --> 00:20:03,680 NONNEURONAL CELL TYPES, SO IF WE 529 00:20:03,680 --> 00:20:05,760 KNOCK OUT GLASS AUTONOMOUSLY IN 530 00:20:05,760 --> 00:20:11,280 THE PIGMENT CELLS WE LOSE THE 531 00:20:11,280 --> 00:20:11,640 RED EYE PIGMENT. 532 00:20:11,640 --> 00:20:14,480 AND IF WE MISS EXPRESS GLASSOT 533 00:20:14,480 --> 00:20:16,720 SIGHTS OR BODY, WE CAN SEE 534 00:20:16,720 --> 00:20:18,360 INDUCTION OF RED EYE OR PIGMENT. 535 00:20:18,360 --> 00:20:19,960 SO GLASS SEEMS TO BE SPECIFIC TO 536 00:20:19,960 --> 00:20:26,760 INDUCE SOME ASPECT OF PIGMENT 537 00:20:26,760 --> 00:20:28,240 CELL IDENTITY. 538 00:20:28,240 --> 00:20:29,680 SO AN TONIA [INDISCERNIBLE] A 539 00:20:29,680 --> 00:20:31,720 MASTER STUDENT WANTED TO KNOW IF 540 00:20:31,720 --> 00:20:33,000 GLASS WAS DIRECTLY REGULATING 541 00:20:33,000 --> 00:20:34,800 THESE GENES THAT ARE SPECIFIC 542 00:20:34,800 --> 00:20:36,320 FOR THE NONNEURONAL CELL TYPES 543 00:20:36,320 --> 00:20:39,600 AND SO SHE TRIED TO CLEAR OUT 544 00:20:39,600 --> 00:20:40,440 SOME ENHANCERS UP STREAM EVER 545 00:20:40,440 --> 00:20:43,520 GENES THAT WE KNEW WERE GLASS 546 00:20:43,520 --> 00:20:45,760 DEPENDENT AND HAD PREDICTED 547 00:20:45,760 --> 00:20:46,480 GLASS BINDING SITES. 548 00:20:46,480 --> 00:20:48,920 AND SHE WAS ABLE TO FIND SOME 549 00:20:48,920 --> 00:20:51,080 THAT WERE SPECIFICALLY EXPRESSED 550 00:20:51,080 --> 00:20:53,120 OR SPECIFICALLY DROVE FOR 551 00:20:53,120 --> 00:20:54,520 REPORTER EXPRESSION IN CONE 552 00:20:54,520 --> 00:20:56,720 CELLS, AND OTHERS THAT 553 00:20:56,720 --> 00:20:58,200 SPECIFICALLY DROVE REPORTER 554 00:20:58,200 --> 00:20:58,800 EXPRESSION IN PIGMENT CELLS. 555 00:20:58,800 --> 00:21:02,000 SO HERE WE SEE THE PRIMARY 556 00:21:02,000 --> 00:21:04,320 PIGMENT CELLS IN THE SECONDARY 557 00:21:04,320 --> 00:21:06,720 AND SERBIARY PIGMENT CELLS AND 558 00:21:06,720 --> 00:21:08,440 THIS EXPRESSION IS GLASS 559 00:21:08,440 --> 00:21:09,960 DEPENDENT BECAUSE IF WE MAKE 560 00:21:09,960 --> 00:21:11,680 GLASS MUTANT CLONES AND THESE 561 00:21:11,680 --> 00:21:13,440 RETINAS AND THE GLASS MUTANT 562 00:21:13,440 --> 00:21:16,520 REGIONS ARE MARKED WITH GFP, WE 563 00:21:16,520 --> 00:21:18,360 SEE LOTS OF REPORTER EXPRESSION 564 00:21:18,360 --> 00:21:21,400 DRIVEN BY THESE ENHANCERS. 565 00:21:21,400 --> 00:21:24,000 SO WE STILL NEED TO MUTATE THESE 566 00:21:24,000 --> 00:21:25,840 GLASS SITES TO SHOW DEFINITIVELY 567 00:21:25,840 --> 00:21:27,720 THAT GLASS IS DIRECTLY 568 00:21:27,720 --> 00:21:28,800 REGULATING THESE ENHANCERS BUT 569 00:21:28,800 --> 00:21:30,120 WE THINK THAT THEY WILL BE 570 00:21:30,120 --> 00:21:32,600 USEFUL TO HELP US UNDERSTAND WHY 571 00:21:32,600 --> 00:21:33,960 GLASS WOULD TURN ON DIFFERENT 572 00:21:33,960 --> 00:21:35,280 GENES IN DIFFERENT CELL TYPES. 573 00:21:35,280 --> 00:21:37,160 SO WHY DOES IT TURN DOWN THIS 1 574 00:21:37,160 --> 00:21:39,240 ONLY IN CONE CELLS, THIS 1 ONLY 575 00:21:39,240 --> 00:21:41,200 IN PIGMENT CELLS AND OTHER GENES 576 00:21:41,200 --> 00:21:43,440 ONLY IF PHOTO RECEPTORS. 577 00:21:43,440 --> 00:21:45,720 AND WE MENTIONED THAT THERE ARE 578 00:21:45,720 --> 00:21:46,640 OTHER TRANSCRIPTION FACTORS THAT 579 00:21:46,640 --> 00:21:49,560 BIND TO THESE ENHANCERS AND 580 00:21:49,560 --> 00:21:53,280 ACCOUNT FOR THESE DIFFERENCES. 581 00:21:53,280 --> 00:21:55,920 AND WE DO KNOW A FEW OF THOSE 582 00:21:55,920 --> 00:21:57,320 TRANSCRIPTION FACTORS JUST BASED 583 00:21:57,320 --> 00:22:00,560 ON OUR GUESSES, SO, FOR 584 00:22:00,560 --> 00:22:03,400 INSTANCE, ESCARGO IS A SPECIFIC 585 00:22:03,400 --> 00:22:05,800 CELL TRANSCRIPTION FACTOR, AND 586 00:22:05,800 --> 00:22:06,560 IT SEEMS TO SYNERGIZE WITH GLASS 587 00:22:06,560 --> 00:22:07,960 TO TURN ON 8 HOURS APPROXIMATE 588 00:22:07,960 --> 00:22:10,320 MENTORSHIP SKILL CELL GENES, SO 589 00:22:10,320 --> 00:22:12,960 IF WE MESS EXPRESSION GLASS, IT 590 00:22:12,960 --> 00:22:16,040 CAN INDUCE RED PIGMENTOT 591 00:22:16,040 --> 00:22:17,560 ABDOMEN, BUT IF WE MISS EXPRESS 592 00:22:17,560 --> 00:22:22,240 GLASS IN CELLS THAT ARE MUTANT 593 00:22:22,240 --> 00:22:23,520 FOR ESCAGOT, IT IS NOT ABLE TO 594 00:22:23,520 --> 00:22:25,320 INDUCE THAT MEG MENTORSHIP SKILL 595 00:22:25,320 --> 00:22:29,440 AND ECTOPIC GLASS IN THE WING 596 00:22:29,440 --> 00:22:31,720 INDUCES ONLY A LITTLE BIT OF 597 00:22:31,720 --> 00:22:34,800 PIGMENT BUT IF WE INDUCE IT WITH 598 00:22:34,800 --> 00:22:37,480 GLASS MORE, WE SEE INDUCTION. 599 00:22:37,480 --> 00:22:42,440 SO WE THINK THAT GLASS AND 600 00:22:42,440 --> 00:22:44,560 ESG CELLS MAY GO TOGETHER AND WE 601 00:22:44,560 --> 00:22:45,720 ALSO IDENTIFIED OTHER 602 00:22:45,720 --> 00:22:47,000 TRANSCRIPTION FACTORS THAT MAY 603 00:22:47,000 --> 00:22:49,680 CONTROL PHOTO REPRESENT 604 00:22:49,680 --> 00:22:51,960 SEPTORSOR SPECIFIC OR CONE CELL 605 00:22:51,960 --> 00:22:54,120 SPECIFIC GENES TOGETHER WITH 606 00:22:54,120 --> 00:22:54,400 GLASS. 607 00:22:54,400 --> 00:22:56,880 SO, THAT'S WHERE WE ARE, WITH 608 00:22:56,880 --> 00:22:57,920 THE CELL FATE QUESTION AND NOW I 609 00:22:57,920 --> 00:22:59,760 WOULD LIKE TO TURN TO THE 610 00:22:59,760 --> 00:23:02,200 QUESTION OF HOW THE CORNEAL LENS 611 00:23:02,200 --> 00:23:06,640 ITSELF IS ACTUALLY PRODUCED BY 612 00:23:06,640 --> 00:23:07,160 THESE CELLS. 613 00:23:07,160 --> 00:23:09,520 SO I REALIZE THIS IS QUITE AN 614 00:23:09,520 --> 00:23:10,880 UNDERSTUDIED AREA, WE ACTUALLY 615 00:23:10,880 --> 00:23:14,160 KNOW VERY LITTLE ABOUT THIS 616 00:23:14,160 --> 00:23:15,440 CORNEAL LENS, AND IT IS A 617 00:23:15,440 --> 00:23:17,160 PARTICULAR STRUCTURE SO WE CAN 618 00:23:17,160 --> 00:23:19,000 GET SOME CLUES FROM HOW IT MIGHT 619 00:23:19,000 --> 00:23:20,760 BE COMPOSED BY OTHER CUTICLES 620 00:23:20,760 --> 00:23:23,480 AND THE LARVA CUTICLE, THE BODY 621 00:23:23,480 --> 00:23:25,040 COVERING, AND THAT CUTICLE IS 622 00:23:25,040 --> 00:23:28,280 MADE UP OF 3 CELL LAYERS, SO 623 00:23:28,280 --> 00:23:30,200 THERE'S AN OUTER LAYER ENVELOPE 624 00:23:30,200 --> 00:23:31,720 THAT HAS WAXES AND PROTEINS, 625 00:23:31,720 --> 00:23:35,640 IT'S A PROTECTIVE LAYER AS IN A 626 00:23:35,640 --> 00:23:37,000 PROCUTICLE THAT CONTAINS THE 627 00:23:37,000 --> 00:23:40,680 CHITIN FIBERS AND IN BETWEEN 628 00:23:40,680 --> 00:23:42,320 THERE'S A EPICUTICLE LAYER 629 00:23:42,320 --> 00:23:43,400 MARKED BY PROTEINS AND OTHERS. 630 00:23:43,400 --> 00:23:45,240 NOW IF WE LOOK AT THESE MARKERS 631 00:23:45,240 --> 00:23:53,680 IN THE CORNEAL LENS, CHITEIN, 632 00:23:53,680 --> 00:23:55,640 SEEMS TO BE ON THE LENS, WE 633 00:23:55,640 --> 00:23:58,240 THINK THAT'S WHERE IT MIGHT BE 634 00:23:58,240 --> 00:24:00,280 ON THE CUTICLE TO BE FOUND BUT 635 00:24:00,280 --> 00:24:04,600 WHEN WE MARKED AN ENDOGENOUS 636 00:24:04,600 --> 00:24:07,200 CUTICLE PROTEIN, WE FOUND IT WAS 637 00:24:07,200 --> 00:24:08,640 IN THE CORNEAL LENS AND NOT IN A 638 00:24:08,640 --> 00:24:09,680 CENTRAL LAYER AS WE THOUGHT IT 639 00:24:09,680 --> 00:24:10,520 MIGHT BE. 640 00:24:10,520 --> 00:24:14,480 SO IT'S REALLY NOT CLEAR, 641 00:24:14,480 --> 00:24:16,320 WHETHER THE CORNEAL LENS HAS THE 642 00:24:16,320 --> 00:24:17,600 SAME POSITION AS THE CUTICLE OR 643 00:24:17,600 --> 00:24:21,080 WHAT THE BULK OF THIS LENS IS 644 00:24:21,080 --> 00:24:25,000 ACTUALLY COMPOSED OF. 645 00:24:25,000 --> 00:24:26,560 SO, WITH THAT IT'S HARD TO TELL 646 00:24:26,560 --> 00:24:28,960 HOW IT'S DEVELOPED, THE 647 00:24:28,960 --> 00:24:30,040 PRECISELY CURVED SHAPE AND OUR 648 00:24:30,040 --> 00:24:37,280 FIRST HANDLE ON THIS CAME WHEN 649 00:24:37,280 --> 00:24:38,960 HONG SUH WAS INVESTIGATING 650 00:24:38,960 --> 00:24:40,040 ANOTHER TRANSCRIPTION FACTOR 651 00:24:40,040 --> 00:24:41,520 CALLED BLIMP 1, QUITE RELATED TO 652 00:24:41,520 --> 00:24:44,800 GLASS AND I SHOULD SAY JAIRED 653 00:24:44,800 --> 00:24:46,640 CALL AT HIS UNIVERSITY AND HIS 654 00:24:46,640 --> 00:24:47,640 OPPORTUNITY DEVELOPED SOME OF 655 00:24:47,640 --> 00:24:49,240 THESE RESULTS AT THE SAME TIME 656 00:24:49,240 --> 00:24:50,960 AND WE COLLABORATED WITH THEM. 657 00:24:50,960 --> 00:24:55,000 SO WHEN THEY KNOCKED OUT BLIMP1 658 00:24:55,000 --> 00:24:58,800 BY MAKING THE MUTATION IN CRSPR, 659 00:24:58,800 --> 00:25:00,880 THEY FOUND THAT BLMP 1 MUTANT 660 00:25:00,880 --> 00:25:02,640 CLONES IN THE ADULT EYE HAD A 661 00:25:02,640 --> 00:25:05,400 RATHER SMOOTH AND GLASSY 662 00:25:05,400 --> 00:25:07,680 SURFACE, WHERE YOU COULDN'T 663 00:25:07,680 --> 00:25:09,480 REALLY DISTINGUISH INDIVIDUAL 664 00:25:09,480 --> 00:25:12,160 CORNEAL LENSS AS YOU CAN IN A 665 00:25:12,160 --> 00:25:12,480 WILD-TYPE EYE. 666 00:25:12,480 --> 00:25:19,560 AND IF YOU CUT SECTIONS THROUGH 667 00:25:19,560 --> 00:25:20,960 THIS EYE, YOU CAN SEE THAT 668 00:25:20,960 --> 00:25:23,360 THROUGH THE WILD-TYPE REGIONS, 669 00:25:23,360 --> 00:25:25,880 THE LENS IS CURVED ON BOTH SIDES 670 00:25:25,880 --> 00:25:28,400 SO IT'S BI CONVEX, BUT IN THE 671 00:25:28,400 --> 00:25:30,040 BLIMP 1 MUTANT REGION WHICH IS 672 00:25:30,040 --> 00:25:31,960 MARK INDEED RED HERE, THE 673 00:25:31,960 --> 00:25:33,640 EXTERNAL SURFACE IS COMPLETELY 674 00:25:33,640 --> 00:25:37,200 FLAT, SO THERE'S A CONVEX LENS. 675 00:25:37,200 --> 00:25:40,400 AND ONCE YOU CALCULATED THE 676 00:25:40,400 --> 00:25:42,000 CHANGE IN FOCAL LENGTH THAT THE 677 00:25:42,000 --> 00:25:43,640 SHAPE CHANGE WOULD CAUSE, AND 678 00:25:43,640 --> 00:25:47,480 FOUND THAT THE FOCAL LENGTH IS 679 00:25:47,480 --> 00:25:48,440 VERY SIGNIFICANTLY INCREASED SO 680 00:25:48,440 --> 00:25:49,840 MUCH SO THAT THESE LENSES WOULD 681 00:25:49,840 --> 00:25:51,280 NOT BE ABLE TO FOCUS LIGHT 682 00:25:51,280 --> 00:25:56,280 WITHIN THE WHOLE THICKNESS OF 683 00:25:56,280 --> 00:25:56,800 THE RETINA. 684 00:25:56,800 --> 00:25:59,680 SO A FLY THAT HAD COMPLETELY 685 00:25:59,680 --> 00:26:01,640 BLIMP 1 EYES WOULD NOT BE ABLE 686 00:26:01,640 --> 00:26:04,760 TO SEE WITH MUCH PRECISION. 687 00:26:04,760 --> 00:26:06,600 SO, WE LOOKEDDA THE WHERE BLIMP 688 00:26:06,600 --> 00:26:08,600 1 IS EXPRESSED AND WE FOUND IT'S 689 00:26:08,600 --> 00:26:10,560 MOST HIGHLY EXPRESS INDEED THE 690 00:26:10,560 --> 00:26:12,720 NONNEURONAL CELL TYPES IN THE 691 00:26:12,720 --> 00:26:15,120 RETINA, THE CONE CELLS PRIMARY 692 00:26:15,120 --> 00:26:17,560 PIGMENT CELLS AND THE SECONDARY 693 00:26:17,560 --> 00:26:19,480 AND TERTIARY PIGMENT CELLS AND 694 00:26:19,480 --> 00:26:21,400 IT HAS A PEEK OF EXPRESSION 695 00:26:21,400 --> 00:26:22,800 ABOUT HALFWAY THROUGH THE PUPIL 696 00:26:22,800 --> 00:26:24,640 STAGE. THIS IS THE TIME AT 697 00:26:24,640 --> 00:26:28,480 WHICH THERE'S A BIG PEEK OF 698 00:26:28,480 --> 00:26:31,760 ECDICEIN, IT'S A STEROID HORMONE 699 00:26:31,760 --> 00:26:33,960 THAT CONTROLS DEVELOPMENTAL TIME 700 00:26:33,960 --> 00:26:34,400 NOTHING DROSOPHILA. 701 00:26:34,400 --> 00:26:39,480 AND WE CAN SHOW THAT BLIMP 1 IS 702 00:26:39,480 --> 00:26:42,000 DEPENDENT ON ECDICEIN, SO IF WE 703 00:26:42,000 --> 00:26:43,280 EXPRESS ECDICEIN RNAi TO KNOCK 704 00:26:43,280 --> 00:26:44,920 DOWN THE SIGNALING IN CLONES IN 705 00:26:44,920 --> 00:26:48,120 THE RETINA MARKED WITH GFP, WE 706 00:26:48,120 --> 00:26:51,560 SAW LOSS OF BLIMP 1 EXPRESSION. 707 00:26:51,560 --> 00:26:53,880 OKAY, SO, IT'S A DEVELOPMENTAL 708 00:26:53,880 --> 00:27:00,720 TIMING SIGNAL IN THESE CELLS. 709 00:27:00,720 --> 00:27:02,000 SO WE THOUGHT THAT FLINT 1 WOULD 710 00:27:02,000 --> 00:27:03,880 BE QUIET IN THE CELLS THAT ARE 711 00:27:03,880 --> 00:27:05,160 KNOWN TO SECRETE MAJOR 712 00:27:05,160 --> 00:27:07,560 COMPONENTS OF THE CORNEAL LENS, 713 00:27:07,560 --> 00:27:08,920 SO THE CURRENT CELLS AND PRIMARY 714 00:27:08,920 --> 00:27:10,520 PEG MENTORSHIP SKILL CELLS ARE 715 00:27:10,520 --> 00:27:11,520 DIRECTLY UPDATERNEATH THE CENTER 716 00:27:11,520 --> 00:27:14,080 OF THE LENS AND THEY ARE KNOWN 717 00:27:14,080 --> 00:27:16,240 TO SECRETE CRYSTALLINE AND OTHER 718 00:27:16,240 --> 00:27:18,240 MAJOR CORNEAL LENS COMPONENTS. 719 00:27:18,240 --> 00:27:20,120 BUT ACTUALLY IF WE KNOCK OUT 720 00:27:20,120 --> 00:27:23,200 BLIMP 1 EXPRESSION SPECIFICALLY 721 00:27:23,200 --> 00:27:26,680 IN THOSE CELLS BY EXPRESSING 722 00:27:26,680 --> 00:27:30,520 BLIMP 1 RANAI FOR THAT'S--RNAi 723 00:27:30,520 --> 00:27:32,080 WITH THE CODE IN THE REGION, 724 00:27:32,080 --> 00:27:35,160 THERE IS NO MAJOR CHANGE IN THE 725 00:27:35,160 --> 00:27:38,840 SHAPE OF THE LENS. 726 00:27:38,840 --> 00:27:41,600 SO WE FOUND THAT THE CELLS THAT 727 00:27:41,600 --> 00:27:43,400 ACTUALLY DO REQUIRE BLIMP 1 TO 728 00:27:43,400 --> 00:27:46,040 CONTROL THE CORNEAL LENS SHAPE 729 00:27:46,040 --> 00:27:47,680 WITH THESE SECONDARY AND 730 00:27:47,680 --> 00:27:49,640 TERTIARY PIGMENT CELLS WHICH ARE 731 00:27:49,640 --> 00:27:51,800 FOUND OF THE PERIPHERY OF 732 00:27:51,800 --> 00:27:53,560 CORNEAL LENS, SO IF WE XOK 733 00:27:53,560 --> 00:27:54,760 KNOCKED DOWN 1 SPHSKLY IN THOSE 734 00:27:54,760 --> 00:27:57,400 CELLS, NOW WE DO SEE CORNEAL 735 00:27:57,400 --> 00:27:59,360 LENSES WITH A FLAT EXTERNAL 736 00:27:59,360 --> 00:28:01,360 SURFACE. 737 00:28:01,360 --> 00:28:02,600 AND YOU CAN SEE THIS. 738 00:28:02,600 --> 00:28:05,560 CLEARLY IF WE DO IT IN THE WHOLE 739 00:28:05,560 --> 00:28:07,800 EYE, SO THE OUTER SURFACE OF THE 740 00:28:07,800 --> 00:28:11,400 LENS IS REALLY COMPLETELY FLAT. 741 00:28:11,400 --> 00:28:13,600 SO WE WERE WONDERING HOW IS IT 742 00:28:13,600 --> 00:28:15,000 THAT THESE CELLS THAT RIGHT AT 743 00:28:15,000 --> 00:28:17,200 THE EDGE OF THE LENS COULD HAVE 744 00:28:17,200 --> 00:28:18,920 AN INFERENCE ON ITS SHAPE AND 1 745 00:28:18,920 --> 00:28:21,360 IDEA WE HAD WAS THAT THESE CELLS 746 00:28:21,360 --> 00:28:23,040 THAT ACTUALLY ATTACH TO THE 747 00:28:23,040 --> 00:28:24,920 PERIPHERY OF THE LENS, AND MAYBE 748 00:28:24,920 --> 00:28:28,480 THEY COULD BE EXERTING SOME KIND 749 00:28:28,480 --> 00:28:30,200 OF MECHANICAL FORCE ON THE LENS 750 00:28:30,200 --> 00:28:33,440 THAT WOULD CHANGE ITS SHAPE. 751 00:28:33,440 --> 00:28:38,000 AND IF WE ZOOM IN ON THIS REGION 752 00:28:38,000 --> 00:28:39,840 WHERE THE PIGMENT CELL ATTACHES 753 00:28:39,840 --> 00:28:42,360 TO THE CORNEAL CELL LENSES AND 754 00:28:42,360 --> 00:28:44,360 LOOK AT THE ELECTRONIC 755 00:28:44,360 --> 00:28:45,680 MICROSCOPY, WE CAN SEE THAT 756 00:28:45,680 --> 00:28:47,400 THERE DOES SEEM TO BE A CHANGE 757 00:28:47,400 --> 00:28:48,960 AT WHICH THE POINTS THAT THE 758 00:28:48,960 --> 00:28:50,920 PIGMENT CELLS ARE LATCHED TO THE 759 00:28:50,920 --> 00:28:52,680 CORNEAL LENS SO IT SEEMS TO BE 760 00:28:52,680 --> 00:28:54,800 MORE OF THESE ATTACHMENT SITES 761 00:28:54,800 --> 00:28:56,240 WHEN WE BEING IMOD DOWN 1 IN THE 762 00:28:56,240 --> 00:28:57,160 PIG M-TEBURKEULOSEIS CELLS SO 763 00:28:57,160 --> 00:28:59,000 THAT MAKE ITS PLAUSIBLE THERE 764 00:28:59,000 --> 00:29:01,840 COULD BE CHANGE IN THE FORCES 765 00:29:01,840 --> 00:29:10,320 THAT APPLY TO THE CORNEAL LENS. 766 00:29:10,320 --> 00:29:11,960 BUT IN ORDER TO GET THESE CELLS, 767 00:29:11,960 --> 00:29:14,360 WE NEED TO KNOW WHAT IS 768 00:29:14,360 --> 00:29:16,000 REGULATING THE PEG MINT IN THE 769 00:29:16,000 --> 00:29:18,760 SECONDARY AND PRIME AREAY CELLS 770 00:29:18,760 --> 00:29:21,240 AND SO SUH DID ANOTHER ANALYSIS 771 00:29:21,240 --> 00:29:23,360 IN WHICH BLIMP 1 WAS KNOCKED 772 00:29:23,360 --> 00:29:24,480 DOWN OR OVEREXPRESSED AND SHE 773 00:29:24,480 --> 00:29:26,000 FOUND MANY DIFFERENT TYPES OF 774 00:29:26,000 --> 00:29:30,000 GENES THAT WERE REGULATED BY 775 00:29:30,000 --> 00:29:32,040 BLIMP 1, BUT 1 CLASS OF GENES 776 00:29:32,040 --> 00:29:35,000 THAT CAUGHT OUR EYE WAS A SET OF 777 00:29:35,000 --> 00:29:40,280 GENES THAT [INDISCERNIBLE] 778 00:29:40,280 --> 00:29:40,680 DOMAIN PROTEINS. 779 00:29:40,680 --> 00:29:43,240 SO SOME OF THESE ARE UPREGULATED 780 00:29:43,240 --> 00:29:44,600 WHEN THESE ARE KNOCKED DOWN AND 781 00:29:44,600 --> 00:29:46,160 OTHERS ARE DOWN REGULATED SO 782 00:29:46,160 --> 00:29:47,000 THESE PROTEINS WERE INTERESTING 783 00:29:47,000 --> 00:29:48,800 BECAUSE THEY WERE KNOWN TO 784 00:29:48,800 --> 00:29:50,480 CONTROL THE SHAPE OF OTHER 785 00:29:50,480 --> 00:29:53,000 CUTICLE STRUCTURES, SUCH AS THE 786 00:29:53,000 --> 00:29:54,000 DENTICLES THAT ARE FOUNDOT 787 00:29:54,000 --> 00:29:57,720 VENTRAL SITE OF THE EMBRYO. 788 00:29:57,720 --> 00:29:58,200 SO [INDISCERNIBLE]'S LAB 789 00:29:58,200 --> 00:30:00,440 RETURNED THAT A NUMBER OF 790 00:30:00,440 --> 00:30:04,120 DIFFERENT CP DOMAIN PROTEINS 791 00:30:04,120 --> 00:30:05,760 EXPRESSED IN THESE DENTICLE 792 00:30:05,760 --> 00:30:07,600 PRODUCING CELLS AND EACH 1 HAS A 793 00:30:07,600 --> 00:30:08,880 DIFFERENT SHAPE ON THE EFFECT OF 794 00:30:08,880 --> 00:30:16,080 THE DEPTICLE WHEN YOU MOVE IT. 795 00:30:16,080 --> 00:30:17,040 SO, [INDISCERNIBLE] TARTED 796 00:30:17,040 --> 00:30:19,720 LOOKAL IF THESE PROTEINS 797 00:30:19,720 --> 00:30:21,480 PHENOTYPES AFFECTING THE SHAPE 798 00:30:21,480 --> 00:30:23,680 OF THE CORNEAL LENS, AND THE 799 00:30:23,680 --> 00:30:24,760 MOST STRIKING 1, SHE'S FOUND SO 800 00:30:24,760 --> 00:30:27,640 FAR IS FOR A GENE CALLED DUSKY 801 00:30:27,640 --> 00:30:29,520 LIKE, OR DYL, AND THIS IS A 802 00:30:29,520 --> 00:30:31,280 PROTEIN THAT'S FOUND AT THE VERY 803 00:30:31,280 --> 00:30:33,400 TIP OF THOSE EMBRYONIC DENTICLES 804 00:30:33,400 --> 00:30:36,360 SO EACH PROTEIN IS LOCALIZED TO 805 00:30:36,360 --> 00:30:42,240 A DIFFERENT REGION OF THE APICAL 806 00:30:42,240 --> 00:30:42,680 EXTRA CELLULAR MATRIX. 807 00:30:42,680 --> 00:30:45,480 SO SHE FOUND WHEN SHE MADE DYL 808 00:30:45,480 --> 00:30:47,440 MUTANT CLONES IN THE RETINA 809 00:30:47,440 --> 00:30:50,120 MARKED BY GFP HERE, THE CORNEAL 810 00:30:50,120 --> 00:30:53,320 LENS IS AGAIN CHANGE THE SHAPE 811 00:30:53,320 --> 00:30:54,840 TO THE EXTERNAL SURFACES PLATTER 812 00:30:54,840 --> 00:31:02,840 AND THE INTERNAL SURFACES MORE 813 00:31:02,840 --> 00:31:03,880 DEEPLY CURVED. 814 00:31:03,880 --> 00:31:05,760 AND WE CAN QUANTIFY THIS BY 815 00:31:05,760 --> 00:31:08,200 MEASURING THE OUTER ANGLE BY 816 00:31:08,200 --> 00:31:10,080 MEASURING 2 ADJACENT CORNEAL 817 00:31:10,080 --> 00:31:13,520 LENSES, SO IN BOTH BLIMP 1 AND 818 00:31:13,520 --> 00:31:14,800 DYL MUTANT REGIONS THAT ANKLE 819 00:31:14,800 --> 00:31:19,240 GOES UP TO ALMOST 180-DEGREES SO 820 00:31:19,240 --> 00:31:20,320 ALMOST COMPLETELY FLAT. 821 00:31:20,320 --> 00:31:23,240 AND THE DYL MUTANT PHENOTYPE CAN 822 00:31:23,240 --> 00:31:25,760 BE RESCUED BY REEXPRESSING DYL 823 00:31:25,760 --> 00:31:29,440 IN THE MUTANT CELLS. 824 00:31:29,440 --> 00:31:32,000 AND I DIDN'T SHOW IT HERE BUT HE 825 00:31:32,000 --> 00:31:33,080 ALWAYS MEASURED THE INNER ANGLE 826 00:31:33,080 --> 00:31:35,160 AND SHOWED THAT IT'S DECREASED 827 00:31:35,160 --> 00:31:39,160 IN THE DYL MUTANT TERMS. 828 00:31:39,160 --> 00:31:42,040 SO TO UNDERSTAND HOW THIS CHANGE 829 00:31:42,040 --> 00:31:43,640 AND CHANGE IN SHAPE IS HAPPENING 830 00:31:43,640 --> 00:31:45,400 I WENT BACK TO EARLIER STAGES OF 831 00:31:45,400 --> 00:31:48,880 DEVELOPMENT, SO SHE LOOKED AT 832 00:31:48,880 --> 00:31:51,880 THE MIDPUPIL RETINA, AND AT THIS 833 00:31:51,880 --> 00:31:54,720 STAGE, THESE CELLS FORM A 834 00:31:54,720 --> 00:31:56,120 BEAUTIFUL REPEATING PATTERN AT 835 00:31:56,120 --> 00:31:57,120 THE APICAL SURFACE OF THE 836 00:31:57,120 --> 00:31:59,120 RETINA, SO YOU SEE 4 CONE CELLS 837 00:31:59,120 --> 00:32:02,920 IN THE MIDDLE, SURROUNDED BY 2 838 00:32:02,920 --> 00:32:04,880 PRIMARY PIGMENT CELLS, AND THEN 839 00:32:04,880 --> 00:32:07,000 SECONDARY PIGMENT CELLS ON EACH 840 00:32:07,000 --> 00:32:08,920 EDGE AND TERTIARY PIGMENT 841 00:32:08,920 --> 00:32:12,160 CELLINGS OR BRISTLES AT THE 842 00:32:12,160 --> 00:32:12,400 CORNERS. 843 00:32:12,400 --> 00:32:14,840 SO WHEN SHE MADE DYL MUTANT 844 00:32:14,840 --> 00:32:18,280 CLONES, IT ARE MARKED WITH GFP 845 00:32:18,280 --> 00:32:20,840 HERE, AND DYL PROTEIN IS MARKED 846 00:32:20,840 --> 00:32:23,440 IN RED, YOU CAN SEE THAT DYL 847 00:32:23,440 --> 00:32:25,280 PROTEIN IS LOCALIZED TO THE 848 00:32:25,280 --> 00:32:27,480 APICKA SURFACES OF THESE CELLS, 849 00:32:27,480 --> 00:32:30,960 AND IN THE DYL MUTANT REGIONS, 850 00:32:30,960 --> 00:32:32,240 THOSE APICAL SURFACES ARE ARE 851 00:32:32,240 --> 00:32:32,480 SMALLER. 852 00:32:32,480 --> 00:32:34,880 SO CAN YOU SEE FOR INSTANCE 853 00:32:34,880 --> 00:32:37,160 HERE, THAT THIS MUTANT PRIMARY 854 00:32:37,160 --> 00:32:38,760 PIGMENT CELL IS SMALLER THAN THE 855 00:32:38,760 --> 00:32:41,440 WILD-TYPE 1 ON THE OTHER SIDE OF 856 00:32:41,440 --> 00:32:43,720 THE [INDISCERNIBLE], AND THE 857 00:32:43,720 --> 00:32:45,480 SAME THING HERE, AND IN OTHER 858 00:32:45,480 --> 00:32:51,240 PLACES AND THE COMPLETELY DYL IS 859 00:32:51,240 --> 00:32:52,000 A SMALLER OVERALL. 860 00:32:52,000 --> 00:32:53,960 AND A FEW HOURS LATER, THIS 861 00:32:53,960 --> 00:32:56,240 EFFECT THAT COMES MUCH MORE 862 00:32:56,240 --> 00:32:59,120 PRONOUNCED SO THE DYL MUTANT 863 00:32:59,120 --> 00:33:04,840 GREENE, OR SIGNIFICANTLY SMALLER 864 00:33:04,840 --> 00:33:05,680 THAN WILD-TYPE MIRROR IMAGE 865 00:33:05,680 --> 00:33:07,880 TIDDIA IN THE RETINA AND THEY'RE 866 00:33:07,880 --> 00:33:10,040 NO LONGER IN THE SAME BASAL 867 00:33:10,040 --> 00:33:17,160 PLAIN AND THAT'S BECAUSE APICKA 868 00:33:17,160 --> 00:33:18,360 CONSTRICTION USUALLY GOES LONG 869 00:33:18,360 --> 00:33:19,800 WITH THE CELLS, SOPHISTICATEDY 870 00:33:19,800 --> 00:33:21,560 IF WE CUT ACROSS SECTION AT THE 871 00:33:21,560 --> 00:33:23,480 RETINA AT A SLIGHTLY LATER 872 00:33:23,480 --> 00:33:26,040 STAGE, YOU CAN SEE THE BASAL 873 00:33:26,040 --> 00:33:28,640 SHORTENING OF THE GFP LABELED 874 00:33:28,640 --> 00:33:31,000 MUTANT CELLS MORE CLIERLY SO 875 00:33:31,000 --> 00:33:32,880 THEY'RE BECOMING SHORTER IN THE 876 00:33:32,880 --> 00:33:34,800 APICKA BASAL THAT I MENTIONED IN 877 00:33:34,800 --> 00:33:36,240 THE NEARBY WILE TYPE CELLS, AND 878 00:33:36,240 --> 00:33:38,680 YOU CAN ALSO SEE, AN 879 00:33:38,680 --> 00:33:40,640 ACCUMULATION OF ACTIN, AT THE 880 00:33:40,640 --> 00:33:45,360 APICAL SURFACE SO THIS IS 881 00:33:45,360 --> 00:33:46,640 LABELED WITH MAURICEIN, AN 882 00:33:46,640 --> 00:33:47,880 ACTIVE BINDING PROTEIN AND IT 883 00:33:47,880 --> 00:33:49,560 CAN ALSO SEE THAT THERE'S A 884 00:33:49,560 --> 00:33:53,520 CHANGE IN THE SHAPE OF THE 885 00:33:53,520 --> 00:33:55,040 DEVELOPING CORNEAL LENS WHICH IS 886 00:33:55,040 --> 00:33:56,760 BULGING MORE INWARD THAN THE 887 00:33:56,760 --> 00:33:58,120 WILD-TYPE LENSES WHICH ARE 888 00:33:58,120 --> 00:33:58,800 BULGING OUTWARD. 889 00:33:58,800 --> 00:34:01,480 SO WE THINK THAT THIS SHORTENING 890 00:34:01,480 --> 00:34:06,040 OF THE OMA TIDIUM WILL PULLOT 891 00:34:06,040 --> 00:34:11,480 LENS TO MECHANICALLY CHANGE ITS 892 00:34:11,480 --> 00:34:11,920 SHAPE. 893 00:34:11,920 --> 00:34:13,240 SO OUR MODEL BASED ON THIS IS 894 00:34:13,240 --> 00:34:15,880 THAT DYL AT THE APICAL SURFACE 895 00:34:15,880 --> 00:34:18,080 OF THESE CONE AND PRIMARY 896 00:34:18,080 --> 00:34:19,320 PIGMENT CELLS MAY BE ATTACHING 897 00:34:19,320 --> 00:34:22,880 IT TO THE DEVELOPING CORNEAL 898 00:34:22,880 --> 00:34:24,760 LENS, AND THAT THESE APICAL 899 00:34:24,760 --> 00:34:27,720 SURFACES WOULD BE EXPANDED AND 900 00:34:27,720 --> 00:34:28,920 KIND OF UNDERTENSION, SO THAT IF 901 00:34:28,920 --> 00:34:32,040 YOU GET RID OF DYL, THOSE CELLS 902 00:34:32,040 --> 00:34:33,640 WOULD SHRINK THE APICKA SURFACES 903 00:34:33,640 --> 00:34:36,480 THAT DETACH FROM THE LENS AND 904 00:34:36,480 --> 00:34:37,600 KIND OF CONTRACT DOWNWARDS. 905 00:34:37,600 --> 00:34:39,920 AND WE THINK THAT THE SECONDARY 906 00:34:39,920 --> 00:34:41,080 AND TERTIARY PIGMENT CELLS ARE 907 00:34:41,080 --> 00:34:42,840 STILL ATTACHED TO THE CORNEAL 908 00:34:42,840 --> 00:34:46,800 LENS AND SO THEY WOULD PULL THE 909 00:34:46,800 --> 00:34:49,360 CORNEAL LENS INWARD AS THE 910 00:34:49,360 --> 00:34:53,880 OMIRROR IMAGE TIDDIA CONTRACT. 911 00:34:53,880 --> 00:34:55,160 SO WE WANTED TO TRY AND TEST 912 00:34:55,160 --> 00:34:59,280 THIS MODEL AND TO DO THAT WE 913 00:34:59,280 --> 00:35:00,440 INDUCED APICAL CONSTRICTION IN 914 00:35:00,440 --> 00:35:01,720 AN ARTIFICIAL WAY THAT HAD 915 00:35:01,720 --> 00:35:04,920 NOTHING TO DO WITH DYL SO SHE 916 00:35:04,920 --> 00:35:07,320 MISS EXPRESSED AN ACTIVATED FORM 917 00:35:07,320 --> 00:35:09,000 OF LIGHT CHAIN KINASE IN CLONES 918 00:35:09,000 --> 00:35:13,080 OF CELLS AND MARKED WITH GFP. 919 00:35:13,080 --> 00:35:15,720 AND YOU CAN SEE THE SAME KIND OF 920 00:35:15,720 --> 00:35:16,480 APICAL CONSTRICTION IN NEEZ 921 00:35:16,480 --> 00:35:18,920 CLONES SO THAT THE APICKA 922 00:35:18,920 --> 00:35:21,240 SURFACES OF THE OMATIDDIA THAT 923 00:35:21,240 --> 00:35:22,960 ARE EXPRESSING IN THE MICE IN 924 00:35:22,960 --> 00:35:26,600 LIGHT CHAIN KINASE ARE SMALLER 925 00:35:26,600 --> 00:35:28,080 THAN NEARBY WILD-TYPE OMATIDDIA, 926 00:35:28,080 --> 00:35:29,800 AND WHEN SHE LOOKEDDA THE ADULTS 927 00:35:29,800 --> 00:35:31,560 THAT DEVELOPED, SHE SAW THAT 928 00:35:31,560 --> 00:35:33,720 AGAIN IN THESE MUTANT REGIONS, 929 00:35:33,720 --> 00:35:36,280 THE SHAPE OF THE CORNEAL LENS 930 00:35:36,280 --> 00:35:38,680 WAS CHANGED, SO AGAIN IT WAS 931 00:35:38,680 --> 00:35:40,840 BULGING INWARD MUCH MORE THAN 932 00:35:40,840 --> 00:35:43,480 THE NEARBY WILD-TYPE REGIONS, SO 933 00:35:43,480 --> 00:35:45,320 APICKA CONSTRUCTION IN ITSELF IS 934 00:35:45,320 --> 00:35:46,360 SUFFICIENT TO CHANGE THE SHAPE 935 00:35:46,360 --> 00:35:52,320 OF THE LENS AND INCREASE THE 936 00:35:52,320 --> 00:35:53,920 INNER CURVEATURE. 937 00:35:53,920 --> 00:35:55,000 SO HOWEVER, THERE'S STILL A 938 00:35:55,000 --> 00:35:56,880 PROBLEM WITH OUR MODEL AND THAT 939 00:35:56,880 --> 00:35:59,520 IS THAT DYL IS NOT THERE FOR 940 00:35:59,520 --> 00:36:03,800 VERY LONG, SO IT HAS THIS VERY 941 00:36:03,800 --> 00:36:06,200 SHARP PEAK OF MRNA SUGGESTION 942 00:36:06,200 --> 00:36:08,840 ACCORDING TO FLY BASE AT THE 943 00:36:08,840 --> 00:36:09,240 MIDPUPIL STAGE. 944 00:36:09,240 --> 00:36:10,440 AND WE CAN SEE THE SAME THING, 945 00:36:10,440 --> 00:36:12,760 IF WE LOOK AT THE PROTEIN SO 946 00:36:12,760 --> 00:36:14,240 IT'S HIGHEST LEVEL OF EXPRESSION 947 00:36:14,240 --> 00:36:19,280 IS AT 50 HOURS OF PUPIL 948 00:36:19,280 --> 00:36:21,280 DEVELOPMENT AND JUST A COUPLE 949 00:36:21,280 --> 00:36:22,200 HOURS LATER THERE'S MUCH LESS 950 00:36:22,200 --> 00:36:24,400 PROTEIN AND THEN IT DISAPPEARS 951 00:36:24,400 --> 00:36:26,280 SO DYL ITSELF CAN'T REALLY BE 952 00:36:26,280 --> 00:36:27,360 FORMING A REALLY LONG LASTING 953 00:36:27,360 --> 00:36:29,000 ASHES TACHMENT OF THE CORNEAL 954 00:36:29,000 --> 00:36:34,280 LENS TO THE UNDERLYING CELLS. 955 00:36:34,280 --> 00:36:36,960 SO WE KNOW THAT CP DOMAIN 956 00:36:36,960 --> 00:36:38,240 PROTEINS CAN POLYMERIZE INTO 957 00:36:38,240 --> 00:36:40,200 LONG FILAMENTS, SO THIS IS 958 00:36:40,200 --> 00:36:43,280 SHARING THE STRUCTURE IF YOU'RE 959 00:36:43,280 --> 00:36:45,160 MODULAR PT PROTEIN AND EM 960 00:36:45,160 --> 00:36:46,880 STRUCTURE OF THOSE FILAMENTS 961 00:36:46,880 --> 00:36:49,320 THAT IT FORMS AND WE ALSO KNOW 962 00:36:49,320 --> 00:36:54,400 THAT DIFFERENT CP PROTEINS CAN 963 00:36:54,400 --> 00:36:54,920 FORM FILAMENTS TOGETHER. 964 00:36:54,920 --> 00:36:57,320 AND SO WE THOUGHT THAT PERHAPS 965 00:36:57,320 --> 00:36:59,360 THE DYL MIGHT AFFECT THE 966 00:36:59,360 --> 00:37:00,440 ORGANIZATION OF OTHER DOMAIN 967 00:37:00,440 --> 00:37:02,200 PROTEINS SO WE LOOKED AT A FEW 968 00:37:02,200 --> 00:37:04,720 OF THEM AND I SAY 1, THAT WAS 969 00:37:04,720 --> 00:37:07,080 KNOWN TO FORM FILAMENTS IS 970 00:37:07,080 --> 00:37:09,160 DUMPY, THIS IS A REALLY ENORMOUS 971 00:37:09,160 --> 00:37:16,960 PROTEIN, THAT HAS A CP DOMAIN, 972 00:37:16,960 --> 00:37:18,640 AND [INDISCERNIBLE] SHARING THAT 973 00:37:18,640 --> 00:37:21,720 AT MUSCLE ATTACHMENT SITES IT 974 00:37:21,720 --> 00:37:23,240 FORMS VERY NICE FILAMENTS THAT 975 00:37:23,240 --> 00:37:25,320 CONNECT THE MUSCLE TO THE 976 00:37:25,320 --> 00:37:25,680 TENDON. 977 00:37:25,680 --> 00:37:27,280 SO [INDISCERNIBLE] FOUND THAT IF 978 00:37:27,280 --> 00:37:30,920 SHE LOOKED AT DUMPING IN THE 979 00:37:30,920 --> 00:37:32,240 CORNEAL LENS IT ALSO SEEMED TO 980 00:37:32,240 --> 00:37:34,240 BE FORMING SOME KIND OF 981 00:37:34,240 --> 00:37:35,880 FILAMENTAL STRUCTURE AND THIS 982 00:37:35,880 --> 00:37:38,760 ORGANIZATION WAS DEPENDENT ON 983 00:37:38,760 --> 00:37:42,240 DYL, SO IN DYL MUTANT REGIONS, 984 00:37:42,240 --> 00:37:46,560 WE DON'T SEE THESE FILAMENTS 985 00:37:46,560 --> 00:37:47,360 FORMING. 986 00:37:47,360 --> 00:37:51,800 AND SHE SAW SIMILAR THING WITH 2 987 00:37:51,800 --> 00:37:55,080 OTHER DOMAIN PROTEINS SO 988 00:37:55,080 --> 00:37:57,680 POPOAGAIN FORMS FILMAMENTS IN 989 00:37:57,680 --> 00:37:59,560 WILD-TYPE REGIONS BUT IN THIS 990 00:37:59,560 --> 00:38:01,080 DYL MUTANT REGIONS AND THE SAME 991 00:38:01,080 --> 00:38:02,920 THING FOR TRINITY, WHICH IS 992 00:38:02,920 --> 00:38:04,280 ANOTHER CP DOMAIN PROTEIN, SO 993 00:38:04,280 --> 00:38:07,560 AGAIN THIS SEEMS TO BE FILAMENTS 994 00:38:07,560 --> 00:38:09,920 IN THE WILD-TYPE BUT NOT IN THE 995 00:38:09,920 --> 00:38:16,240 DYL MUTANT REGIONS OF THE 996 00:38:16,240 --> 00:38:16,480 RETINA. 997 00:38:16,480 --> 00:38:17,960 SO THIS MAY BE PART OF THE 998 00:38:17,960 --> 00:38:21,000 ANSWER TO WHAT DYL IS DOING BUT 999 00:38:21,000 --> 00:38:21,600 IT'S--LIKELY MORE COMPLICATED 1000 00:38:21,600 --> 00:38:24,600 THAN THAT BECAUSE IF WE ELECTRIC 1001 00:38:24,600 --> 00:38:25,960 AT ANOTHER NONCP DOMAIN OPPOSITE 1002 00:38:25,960 --> 00:38:29,840 BEHAVIORIAL PHENOTYPENT AS THE 1003 00:38:29,840 --> 00:38:31,320 CORNEAL LENS SO THE 1004 00:38:31,320 --> 00:38:33,080 [INDISCERNIBLE] LENS THAT IS 1005 00:38:33,080 --> 00:38:34,960 FOUND AT THE PERIPHERY OF THE 1006 00:38:34,960 --> 00:38:35,640 LENS, THAT DOESN'T SEEM FOCUSED 1007 00:38:35,640 --> 00:38:38,840 ON BE A SECRETED OR DEPOSITED 1008 00:38:38,840 --> 00:38:49,320 PROPERLY IN MUTANT CLONES SO 1009 00:38:51,520 --> 00:38:52,480 IT'S IS STUCK IN THE ZONES. 1010 00:38:52,480 --> 00:38:54,800 SO WE THINK THAT DYL IS HELPING 1011 00:38:54,800 --> 00:39:01,040 TO FORM A NETWORK OF CPL PROTEIN 1012 00:39:01,040 --> 00:39:02,160 FILAMENTS INCLUDING PPOT TRINITY 1013 00:39:02,160 --> 00:39:05,160 AND IT MAY ALSO BE FORMING A 1014 00:39:05,160 --> 00:39:07,040 CONDUIT FOR THE DEPOSITION OF 1015 00:39:07,040 --> 00:39:08,280 OTHER COMOPPOSITE BEHAVIORIAL 1016 00:39:08,280 --> 00:39:15,040 PHENOTYPENTS SUCH AS SWEDELE E. 1017 00:39:15,040 --> 00:39:18,360 OKAY SO THAT'S ALL WELL AND GOOD 1018 00:39:18,360 --> 00:39:20,560 BUT WE HAVEN'T COMPLETELY SOLVED 1019 00:39:20,560 --> 00:39:22,040 THE PROBLEM OF THE THIS YET 1020 00:39:22,040 --> 00:39:24,840 BECAUSE EVEN THOUGH IT'S 1021 00:39:24,840 --> 00:39:27,040 REGULATED BY DYL 1, IT DOESN'T 1022 00:39:27,040 --> 00:39:29,440 ACCOUNT FOR THE LENGTH OF 1 ON 1023 00:39:29,440 --> 00:39:29,840 THE CORNEAL SHAPE. 1024 00:39:29,840 --> 00:39:33,440 BECAUSE IF WE LOOK AT BLIMP 1 1025 00:39:33,440 --> 00:39:35,280 CLEARANCE THE SAME STAGE, THE 1026 00:39:35,280 --> 00:39:38,480 BLIMP 1 MUTANT CELLS WHICH ARE 1027 00:39:38,480 --> 00:39:41,040 MARKED IN RED HERE EXPAND THE 1028 00:39:41,040 --> 00:39:44,040 IPICKAL PROFILES SO THAT BIGGER 1029 00:39:44,040 --> 00:39:48,080 THAN NEIGHBORING WILD-TYPE CELLS 1030 00:39:48,080 --> 00:39:50,160 AND ALSO RATHER THAN JUST 1031 00:39:50,160 --> 00:39:52,840 CHANGING, IT DOESN'T SEEM TO 1032 00:39:52,840 --> 00:39:54,360 SECRETE MUCH DUMPY AT ALL. 1033 00:39:54,360 --> 00:39:57,280 SO BLIMP 1 IS DOING OTHER THINGS 1034 00:39:57,280 --> 00:40:02,080 HAN JUST REGULATING DYL. 1035 00:40:02,080 --> 00:40:04,240 SO 1 CLASS OF BLIMP 1 TARGET 1036 00:40:04,240 --> 00:40:05,400 GENES THAT WE'RE INTERESTED IN 1037 00:40:05,400 --> 00:40:07,160 LOOKING AT IS WHETHER THOSE ARE 1038 00:40:07,160 --> 00:40:10,000 IMPRESSED BY BLIMP 1. 1039 00:40:10,000 --> 00:40:13,200 SO BLIMP 1 IS KNOWN TO ACT AS A 1040 00:40:13,200 --> 00:40:16,320 SUPPRESSOR IN SOME OTHER 1041 00:40:16,320 --> 00:40:17,080 SYSTEMS. 1042 00:40:17,080 --> 00:40:18,920 SO IN BLIMP 1 NUCLEOTIDESITANT 1043 00:40:18,920 --> 00:40:21,240 CELLS THESE REPRESSED TARGETS 1044 00:40:21,240 --> 00:40:23,200 WOULD BE UPREGULATED AND WE 1045 00:40:23,200 --> 00:40:27,280 REASONED THAT IF WE WERE TO DOWN 1046 00:40:27,280 --> 00:40:28,360 REGULATE THOSE MESS EXPRESSED 1047 00:40:28,360 --> 00:40:29,640 PROTEINS WE MIGHT BE ABLE TO 1048 00:40:29,640 --> 00:40:32,280 RESCUE THE FINE O TYPE OF BLIMP 1049 00:40:32,280 --> 00:40:32,600 1. 1050 00:40:32,600 --> 00:40:36,720 AND YOU KNOW THIS WAS PROBABLY, 1051 00:40:36,720 --> 00:40:38,120 TOO SIMPLISTIC BECAUSE THERE'S 1052 00:40:38,120 --> 00:40:40,040 SO MANY TARGETS OF BLIMP 1, IT 1053 00:40:40,040 --> 00:40:41,600 WAS PERHAPS UNLIKELY WE WOULD 1054 00:40:41,600 --> 00:40:43,280 FIND A SIPPING SINGLE 1 THAT 1055 00:40:43,280 --> 00:40:44,560 WOULD RESCUE THE PHENOTYPE BUT 1056 00:40:44,560 --> 00:40:45,840 WE ACTUALLY DID FIND 1 THAT HAD 1057 00:40:45,840 --> 00:40:48,320 A PRETTY STRONG EFFECT. 1058 00:40:48,320 --> 00:40:51,320 SO THIS SAY PROTEIN CALLED 1059 00:40:51,320 --> 00:40:51,560 CT433. 1060 00:40:51,560 --> 00:40:53,960 WE HAVE TO GIVE IT A BETTER NAME 1061 00:40:53,960 --> 00:40:58,040 SOON BUT IT WAS UPREGULATED IN 1062 00:40:58,040 --> 00:41:01,040 BLIMP 1 KNOCK DOWN RETINAS AND 1063 00:41:01,040 --> 00:41:04,240 IT SEEMS TO RESCUE THE PHENOTYPE 1064 00:41:04,240 --> 00:41:06,600 SO AGAIN BLIMP 1 CLONES HAVE 1065 00:41:06,600 --> 00:41:09,400 FLAT CORNEAL LENSES, BUT IF WE 1066 00:41:09,400 --> 00:41:10,600 MAKE BLIMP 1 MUTANT CLONES IN 1067 00:41:10,600 --> 00:41:13,400 WHICH WE KNOCK DOWN CT4333, THEY 1068 00:41:13,400 --> 00:41:16,880 HAVE MUCH MORE NORMAL CURVEATURE 1069 00:41:16,880 --> 00:41:18,280 FROM THE OUTSIDE, THE EYE 1070 00:41:18,280 --> 00:41:19,800 DOESN'T HAVE THOSE SMOOTH GLASSY 1071 00:41:19,800 --> 00:41:22,640 PATCHES, AND IF WE MEASURE THAT 1072 00:41:22,640 --> 00:41:24,160 OUTER LENS ANGLE IT'S RESTORED 1073 00:41:24,160 --> 00:41:29,560 TO A MORE NORMAL ANGLE. 1074 00:41:29,560 --> 00:41:36,200 AND THIS WAS ESPECIALLY 1075 00:41:36,200 --> 00:41:38,600 INTERESTING BECAUSE CT4333, IS 1076 00:41:38,600 --> 00:41:41,600 USED TO PROMOTE GTFBI, GIVE RISE 1077 00:41:41,600 --> 00:41:48,280 TO MANY HUMAN CORNEAL 1078 00:41:48,280 --> 00:41:48,600 DYSTROPHIES. 1079 00:41:48,600 --> 00:41:51,560 SO TGFBI AND CT4333 ARE BOTH 1080 00:41:51,560 --> 00:41:53,480 CLASSIC DOMAIN PROTEINS AND 1081 00:41:53,480 --> 00:41:55,680 WITHIN THOSE CLASSIC DOMAINS 1082 00:41:55,680 --> 00:41:58,040 THERE ARE MANY AMINO ACIDS THAT 1083 00:41:58,040 --> 00:41:59,160 ARE BOTH CONSERVED BETWEEN THESE 1084 00:41:59,160 --> 00:42:01,720 2 PROTEINS AND CAN MUTATE TO 1085 00:42:01,720 --> 00:42:04,920 GIVE HUMAN CORNEAL DYSTROPHY. 1086 00:42:04,920 --> 00:42:06,920 AND OUR TECHNICIAN IN THE LAB IS 1087 00:42:06,920 --> 00:42:08,760 NOW WORKING TO KNOCK IN SOME OF 1088 00:42:08,760 --> 00:42:10,400 THESE POINT MUTATIONS THAT GIVE 1089 00:42:10,400 --> 00:42:12,720 CORNEAL DYSTROPHY IN THE CT4333 1090 00:42:12,720 --> 00:42:17,920 TO SEE IF WE CAN GET AN EYE FINE 1091 00:42:17,920 --> 00:42:18,520 O TYPE. 1092 00:42:18,520 --> 00:42:22,040 BUT ANOTHER THING WE DO KNOW IS 1093 00:42:22,040 --> 00:42:23,520 THAT MANY OF THESE MUTATIONS 1094 00:42:23,520 --> 00:42:25,160 THAT GIVE RISE TO CORNEAL 1095 00:42:25,160 --> 00:42:26,760 DYSTROPHY HAVE THAT EFFECT 1096 00:42:26,760 --> 00:42:28,360 BECAUSE THEY CAUSE ACCUMULATION 1097 00:42:28,360 --> 00:42:30,120 OR AGGREGATION OF THE PROTEIN, 1098 00:42:30,120 --> 00:42:35,080 SO, THEY CAN EITHER EFFECT ITS 1099 00:42:35,080 --> 00:42:39,240 SOLUBLE OR TURNOVER BY UBIQUITIN 1100 00:42:39,240 --> 00:42:39,960 MEDIATION OR DISINDUCED 1101 00:42:39,960 --> 00:42:42,600 AGGREGATION OF THE PROTEIN AND 1102 00:42:42,600 --> 00:42:45,600 CAUSES ACCUMULATION OF THE 1103 00:42:45,600 --> 00:42:46,000 CORNEAL DYSTROPHY. 1104 00:42:46,000 --> 00:42:50,520 AND SO, WE WANTED TO SEE WHETHER 1105 00:42:50,520 --> 00:42:51,440 JUST OVEREXPRESSING CT4333 MIGHT 1106 00:42:51,440 --> 00:42:54,280 HAVE AN EFFECT AND IN FACT IT 1107 00:42:54,280 --> 00:42:56,000 DOES APPEAR TO FLATTEN THE 1108 00:42:56,000 --> 00:42:58,320 EXTERNAL SURFACE OF THE LENS. 1109 00:42:58,320 --> 00:43:01,920 SO WE'RE HOPEFUL THAT THERE 1110 00:43:01,920 --> 00:43:04,240 MIGHT BE MORE HOMOLOGY BETWEEN 1111 00:43:04,240 --> 00:43:06,520 THE CORNEAL LENS OF THE HUMAN 1112 00:43:06,520 --> 00:43:07,720 CORNEA THAN MIGHT A--BITS PEESH 1113 00:43:07,720 --> 00:43:09,160 AT FIRST GLASS. 1114 00:43:09,160 --> 00:43:10,760 AND THAT THINKING ABOUT 1115 00:43:10,760 --> 00:43:13,000 CONSERVATION MADE US GO BACK AND 1116 00:43:13,000 --> 00:43:16,280 LOOK AT BLIMP 1 BECAUSE THE 1117 00:43:16,280 --> 00:43:17,480 [INDISCERNIBLE] LAB IN BLIMP 1 1118 00:43:17,480 --> 00:43:19,000 IS EXPRESSED IN THE HUMAN 1119 00:43:19,000 --> 00:43:20,640 CORNEA, THIS IS MOUSE CORNEA, 1120 00:43:20,640 --> 00:43:25,360 SORRY, BUT IT'S SPECIFICALLY 1121 00:43:25,360 --> 00:43:26,840 LOCALIZED TO THE LIMBUS AND THE 1122 00:43:26,840 --> 00:43:28,200 THAT'S THE REGION WHERE THE 1123 00:43:28,200 --> 00:43:30,120 SURVEATURE OF THE CORNEA 1124 00:43:30,120 --> 00:43:32,680 INITIATES, IT'S ALSO WELL ADULT 1125 00:43:32,680 --> 00:43:35,880 EPITHELIAL STEM CELLS ARE FOUND. 1126 00:43:35,880 --> 00:43:37,920 SO, WE ARE INTERESTED IN 1127 00:43:37,920 --> 00:43:39,800 LEARNING MORE ABOUT THE FUNCTION 1128 00:43:39,800 --> 00:43:41,880 OF BLIMP 1 IN THE CORNEA, AND SO 1129 00:43:41,880 --> 00:43:45,200 WE STARTED TO COLLABORATION WITH 1130 00:43:45,200 --> 00:43:46,800 CHECKING AT NYU, AND SHE'S 1131 00:43:46,800 --> 00:43:51,120 INTERESTED IN DISEASES OF 1132 00:43:51,120 --> 00:43:52,600 CORNEAL CURVEATURE SUCH AS 1133 00:43:52,600 --> 00:43:53,600 [INDISCERNIBLE] IN WHICH THE 1134 00:43:53,600 --> 00:43:55,440 CORNEA IS TOO CURVED AND SHE 1135 00:43:55,440 --> 00:43:57,840 USES BOTH MICE AND HUMAN CORNEAL 1136 00:43:57,840 --> 00:43:59,000 ORGANNITES IN ORDER TO STUDY 1137 00:43:59,000 --> 00:44:00,040 THIS SO WE THOUGHT THIS WOULD BE 1138 00:44:00,040 --> 00:44:02,000 A GREAT SYSTEM IN WHICH WE CAN 1139 00:44:02,000 --> 00:44:05,800 LOOK AT THE FUNCTION OF BLIMP 1. 1140 00:44:05,800 --> 00:44:08,080 SO WE HAVEN'T GOT VERY FAR WITH 1141 00:44:08,080 --> 00:44:09,840 THIS BUT SHE HAS FOUND THAT FROM 1142 00:44:09,840 --> 00:44:12,080 HER SINGLE CELL RNA SEQ ANALYSIS 1143 00:44:12,080 --> 00:44:16,280 OF BOTH HUMAN CORNEA, AND HUMAN 1144 00:44:16,280 --> 00:44:17,040 CORNEAL ORGANOIDS THAT BLIMP 1 1145 00:44:17,040 --> 00:44:19,400 IS EXPRESSED IN A SUBSET OF THE 1146 00:44:19,400 --> 00:44:21,600 EPITHELIAL CELLS, IN BOTH THE 1147 00:44:21,600 --> 00:44:27,000 CORNEAS AND THE ORGANOIDS. 1148 00:44:27,000 --> 00:44:28,440 AND HER POST DOC [INDISCERNIBLE] 1149 00:44:28,440 --> 00:44:31,440 SHAZ SHATTERED TO LOOK AT BLIMP 1150 00:44:31,440 --> 00:44:34,200 1 TARGET GENES BEGINNING WITH A 1151 00:44:34,200 --> 00:44:35,320 HUMAN CORNEAL EPITHELIAL CELL 1152 00:44:35,320 --> 00:44:36,720 LINE AND THEY'RE STILL WAITING 1153 00:44:36,720 --> 00:44:39,400 FOR THE GENOME WIDE ANALYSIS, 1154 00:44:39,400 --> 00:44:42,080 BUT HE HAS FOUND AT LEAST 1 GENE 1155 00:44:42,080 --> 00:44:44,440 THAT'S REGULATED BY BLIMP 1 IN 1156 00:44:44,440 --> 00:44:46,720 BOTH SYSTEMS WHICH IS 1157 00:44:46,720 --> 00:44:48,080 [INDISCERNIBLE] AND THAT'S 1158 00:44:48,080 --> 00:44:50,160 ENCODED BY HSPT2 IN THE HUMAN 1159 00:44:50,160 --> 00:44:52,160 CORNEA AND THAT'S REDUCED WHEN 1160 00:44:52,160 --> 00:44:56,920 HE KNOCKS DOWN BLIMP 1 WITH 1161 00:44:56,920 --> 00:45:00,120 SiRNA, AND THE GENE IS TROLL, 1162 00:45:00,120 --> 00:45:01,960 AND WE ALSO SAW A RIDUKS IN 1163 00:45:01,960 --> 00:45:03,440 TROLL WHEN WE KNOCKED DOWN BLIMP 1164 00:45:03,440 --> 00:45:05,520 1 IN THE FLY RETINAMENT SO WE 1165 00:45:05,520 --> 00:45:06,880 ARE HOPING THERE WILL BE MORE 1166 00:45:06,880 --> 00:45:09,160 COMMON TARGETS THAT WILL HELP US 1167 00:45:09,160 --> 00:45:12,480 UNDERSTAND WHETHER WE CAN USE 1168 00:45:12,480 --> 00:45:17,320 THE CORNEAL LENS TO MODEL THE 1169 00:45:17,320 --> 00:45:19,600 HUMAN CORNEA. 1170 00:45:19,600 --> 00:45:21,680 SO I HOPE THAT--IT'S STILL EARLY 1171 00:45:21,680 --> 00:45:24,880 DAYS BUT WE DO HOPE THAT WE CAN 1172 00:45:24,880 --> 00:45:26,760 USE THIS CORNEAL LENS, YOU KNOW 1173 00:45:26,760 --> 00:45:27,800 NOT ONLY TO UNDERSTAND HOW WE 1174 00:45:27,800 --> 00:45:30,120 CAN MAKE A CURVED STRUCTURE OUT 1175 00:45:30,120 --> 00:45:32,720 OF EXTRA CELLULAR MATRIX, BUT 1176 00:45:32,720 --> 00:45:34,720 ALSO HOW THE CURVEATURE OF THE 1177 00:45:34,720 --> 00:45:36,360 HUMAN CORNEA IS GENERATED AND 1178 00:45:36,360 --> 00:45:38,840 WE'RE HOPING TO FIND SOME MORE 1179 00:45:38,840 --> 00:45:42,320 COMMON FEATURES, PERHAPS IT WILL 1180 00:45:42,320 --> 00:45:45,000 TURN OUT THAT TGF BI IN THE 1181 00:45:45,000 --> 00:45:47,760 CORNEA IS REPRESSED BY BLIMP 1. 1182 00:45:47,760 --> 00:45:50,800 THERE MIGHT BE RULES FOR 1183 00:45:50,800 --> 00:45:51,920 CP-DOMAIN PROTEINS PERHAPS AS CP 1184 00:45:51,920 --> 00:45:53,720 IS EXPRESS INDEED THE CORNEAL 1185 00:45:53,720 --> 00:45:54,760 ENDOTHELIUM AND IT WILL BE 1186 00:45:54,760 --> 00:45:56,040 INTERESTING IF IT TURNED OUT TO 1187 00:45:56,040 --> 00:46:00,480 ALSO HAVE A ROLE IN ATTACHMENT 1188 00:46:00,480 --> 00:46:02,320 OF THE CORNEA, AND YOU KNOW 1189 00:46:02,320 --> 00:46:04,440 WE'RE LOOKING FORWARD TO 1190 00:46:04,440 --> 00:46:05,120 INVESTIGATING, POSSIBLY 1191 00:46:05,120 --> 00:46:09,880 CONSERVED MECHANISMS IN CORNEAL 1192 00:46:09,880 --> 00:46:10,560 DEVELOPMENT AND DISEASE. 1193 00:46:10,560 --> 00:46:11,280 SOPHISTICATED WITH THAT I WOULD 1194 00:46:11,280 --> 00:46:12,640 LIKE TO THANK MY LAB MEMBERS SO 1195 00:46:12,640 --> 00:46:21,000 I TALKED ABOUT THE WORK OF 1196 00:46:21,000 --> 00:46:26,800 [LISTING STUDENTS ] AND OUR 1197 00:46:26,800 --> 00:46:27,320 COLLABORATORS] LISTING 1198 00:46:27,320 --> 00:46:28,320 COLLABORATORS ] AND OF COURSE I 1199 00:46:28,320 --> 00:46:30,120 WOULD REALLY LIKE TO THANK NIH 1200 00:46:30,120 --> 00:46:32,760 FOR FUNDING THESE PROGECS. 1201 00:46:32,760 --> 00:46:35,400 SO THANK YOU. 1202 00:46:35,400 --> 00:46:39,400 [ APPLAUSE ] 1203 00:46:39,400 --> 00:46:41,280 >>--THAT WAS QUITE A STORY. 1204 00:46:41,280 --> 00:46:51,320 I GOT THE FEELING THAT 1205 00:46:51,320 --> 00:46:52,080 [INDISCERNIBLE] IS A PROTEIN 1206 00:46:52,080 --> 00:46:54,120 THAT MUST BE TIGHTLY REGULATED. 1207 00:46:54,120 --> 00:46:56,560 TOO MUCH AND TOO MUCH CAN CREATE 1208 00:46:56,560 --> 00:46:59,120 HAVOC IN FIBER FORMATION OF THE 1209 00:46:59,120 --> 00:47:00,720 OTHER ZONE [INDISCERNIBLE] 1210 00:47:00,720 --> 00:47:03,680 PROTEINS, I WAS WONDERING YOU 1211 00:47:03,680 --> 00:47:05,800 SHOWED US HOW THE LOSS OF 1212 00:47:05,800 --> 00:47:09,120 FUNCTION CREATE MORE 1213 00:47:09,120 --> 00:47:13,560 CONSTRICTION, COULD WE ASSUME 1214 00:47:13,560 --> 00:47:16,760 THAT OVEREXPRESSION OF IT WILL 1215 00:47:16,760 --> 00:47:17,040 RELAX? 1216 00:47:17,040 --> 00:47:17,840 >>SO THAT'S INTERESTING. 1217 00:47:17,840 --> 00:47:19,080 THAT IS SOMETHING THAT WE SHOULD 1218 00:47:19,080 --> 00:47:21,640 PROBABLY LOOK AT SO WE HAVEN'T 1219 00:47:21,640 --> 00:47:24,920 REALLY DONE MUCH ANALYSIS OF DYL 1220 00:47:24,920 --> 00:47:25,960 OVEREXPRESSION BUT THAT WOULD 1221 00:47:25,960 --> 00:47:27,040 CERTAINLY BE INTERESTING TO LOOK 1222 00:47:27,040 --> 00:47:28,200 AT BECAUSE IT DOES SEEM TO BE 1223 00:47:28,200 --> 00:47:30,360 EXTREMELY TIGHTLY REGULATED. 1224 00:47:30,360 --> 00:47:31,880 SO, YEAH, THANKS. 1225 00:47:31,880 --> 00:47:33,840 >>THANK YOU FOR YOUR VERY NICE 1226 00:47:33,840 --> 00:47:34,760 AND VERY CLEAR TALK. 1227 00:47:34,760 --> 00:47:36,760 THIS IS NOT MY AREA OF EXPERTISE 1228 00:47:36,760 --> 00:47:39,200 BUT I WAS WONDING YOU TALKED A 1229 00:47:39,200 --> 00:47:41,360 LOT ABOUT HOW THESE PROTEIN 1230 00:47:41,360 --> 00:47:43,880 SHAPED THE CORNEA DURING REGULAR 1231 00:47:43,880 --> 00:47:44,880 DEVELOPMENT, DO THEY CHANGE IN 1232 00:47:44,880 --> 00:47:46,320 THE ADULT AFTER DEVELOPMENT 1233 00:47:46,320 --> 00:47:47,280 BECAUSE OBVIOUSLY THERE SEEM TO 1234 00:47:47,280 --> 00:47:50,280 BE CHANGES IN THE CORNEAL 1235 00:47:50,280 --> 00:47:51,760 STRUCTURE IN HUMANS ANYHOW AFTER 1236 00:47:51,760 --> 00:47:55,080 BIRTH AND SO, ARE THIS CHANGES 1237 00:47:55,080 --> 00:47:59,200 IN THESE PROTEINS IN THE ADULT 1238 00:47:59,200 --> 00:47:59,360 FLY. 1239 00:47:59,360 --> 00:48:00,520 >>YEAH, THAT'S ANOTHER THING 1240 00:48:00,520 --> 00:48:03,720 THAT WE HAVEN'T REALLY LOOKED AT 1241 00:48:03,720 --> 00:48:05,400 MUCH YET, SO, I DON'T THINK 1242 00:48:05,400 --> 00:48:07,600 ANYONE HAS MUCH STUDIED HOW THE 1243 00:48:07,600 --> 00:48:08,720 CORNEAL LENS MIGHT CHANGE WITH 1244 00:48:08,720 --> 00:48:11,080 AGE BECAUSE IT IS JUST THIS 1245 00:48:11,080 --> 00:48:13,160 COMPLETELY ACELLULAR, EXTRA 1246 00:48:13,160 --> 00:48:13,960 CELLULAR MATRIX STRUCTURE. 1247 00:48:13,960 --> 00:48:16,040 AND SO I DON'T KNOW HOW MUCH 1248 00:48:16,040 --> 00:48:18,280 TURNOVER THERE IS OF THE 1249 00:48:18,280 --> 00:48:20,040 COMPONENTS AND WHETHER THEY KEEP 1250 00:48:20,040 --> 00:48:22,080 BEING DEPOSITED IN THE ADULT OR 1251 00:48:22,080 --> 00:48:23,280 NOT, THAT IS SOMETHING THAT 1252 00:48:23,280 --> 00:48:26,400 WE'VE CONSIDERED LOOKING AT WITH 1253 00:48:26,400 --> 00:48:29,320 THIS CT433, 3 MUTATIONS BECAUSE 1254 00:48:29,320 --> 00:48:31,160 CORNEAL DYSTROPHY IS SOMETHING 1255 00:48:31,160 --> 00:48:33,560 THAT'S DEFINITELY AN AGE RELATED 1256 00:48:33,560 --> 00:48:36,560 DISORDER THAT GETS WORSE WITH 1257 00:48:36,560 --> 00:48:37,880 AGE AND DIFFERENT AGE HAS 1258 00:48:37,880 --> 00:48:40,080 DIFFERENT ONSETS IN HUMANS SO 1259 00:48:40,080 --> 00:48:41,200 HERE WE ARE CONSIDERING AGING 1260 00:48:41,200 --> 00:48:42,720 THOSE FLIES ONCE WE MAKE THE 1261 00:48:42,720 --> 00:48:46,760 MUTATIONS AND SEEING IF AGE HAS 1262 00:48:46,760 --> 00:48:47,840 ANY PHENOTYPE. 1263 00:48:47,840 --> 00:48:48,640 NTHAT WAS A FANTASTIC TALK. 1264 00:48:48,640 --> 00:48:50,880 I HAD A COUPLE QUESTIONS ABOUT 1265 00:48:50,880 --> 00:48:53,720 YOUR GLASS BINDING RESULTS. 1266 00:48:53,720 --> 00:48:55,280 ONE OF THEM WAS DOES THE LEG 1267 00:48:55,280 --> 00:48:56,600 EXPRESS THE CO FACTOR THAT'S IN 1268 00:48:56,600 --> 00:48:59,000 THE PIGMENT CELLS FOR THE 1269 00:48:59,000 --> 00:49:00,880 PIGMENT SPECIFIC GENES IS THAT 1270 00:49:00,880 --> 00:49:02,720 WHY THE TRICK OF THE GLASS WORKS 1271 00:49:02,720 --> 00:49:04,240 TO TURN ON THE PIG 1272 00:49:04,240 --> 00:49:04,760 M-TEBURKEULOSEIS GENES? 1273 00:49:04,760 --> 00:49:06,160 AND THE RPGHT 1 IS I THINK YOU 1274 00:49:06,160 --> 00:49:07,560 HAD SEVERAL GLASS TARGETS SHARED 1275 00:49:07,560 --> 00:49:10,200 ACROSS ALL CELL TYPES DO YOU 1276 00:49:10,200 --> 00:49:12,080 THINK THERE'S A UBIQUITOUSLY 1277 00:49:12,080 --> 00:49:13,000 EXPRESSED CO FACTOR ARE FOR 1278 00:49:13,000 --> 00:49:14,080 THOSE GENES OR A DIFFERENT IN 1279 00:49:14,080 --> 00:49:15,840 THE NATURE OF THE BINDING SITES 1280 00:49:15,840 --> 00:49:17,440 THAT LETS GLASS BIND ON ITS OWN 1281 00:49:17,440 --> 00:49:20,440 SOMETIMES BUT NEED A CO FACTOR 1282 00:49:20,440 --> 00:49:20,840 OTHER TIMES. 1283 00:49:20,840 --> 00:49:25,960 >>THOSE ARE GREAT QUESTIONS, SO 1284 00:49:25,960 --> 00:49:27,000 ESG IS THE PIGMENT CELL 1285 00:49:27,000 --> 00:49:28,000 TRANSCRIPTION FACTOR I MENTIONED 1286 00:49:28,000 --> 00:49:31,840 SO THAT IS EXPRESSED IN THE LEG. 1287 00:49:31,840 --> 00:49:37,240 IT'S MOSTLY IN THE MOST PROXIMAL 1288 00:49:37,240 --> 00:49:38,560 LEG, BUT IT DOES VERY EARLY ON, 1289 00:49:38,560 --> 00:49:40,320 IT WOULD BE IN THE LARGER REGION 1290 00:49:40,320 --> 00:49:42,040 OF THE LEG DISK AND THAT MAY BE 1291 00:49:42,040 --> 00:49:43,920 WHY WE DON'T SEE THIS MEG 1292 00:49:43,920 --> 00:49:45,040 MENTORSHIP SKILL NECESSARILY 1293 00:49:45,040 --> 00:49:47,120 RESTRICTED TO THE MOST PROXIMAL 1294 00:49:47,120 --> 00:49:48,840 PART OF THE LEG, AND YEAH, AS 1295 00:49:48,840 --> 00:49:50,160 FOR THE GENES THAT REGULATE THE 1296 00:49:50,160 --> 00:49:51,720 GLASS IN ALL CELL TYPES WE 1297 00:49:51,720 --> 00:49:52,920 DEFINITELY LOOK MORE CLOSELY AT 1298 00:49:52,920 --> 00:49:54,120 WHAT THEY ARE, WE'RE JUST 1299 00:49:54,120 --> 00:50:00,080 GETTING THESE DATA RIGHT NOW AND 1300 00:50:00,080 --> 00:50:02,120 SEE WHETHER--SO I DON'T KNOW 1301 00:50:02,120 --> 00:50:03,280 WHETHER OTHER TRANSCRIPTION 1302 00:50:03,280 --> 00:50:04,320 FACTORS WOULD BE NECESSARY TO 1303 00:50:04,320 --> 00:50:05,600 BIND GLASS OR WHETHER THEY WOULD 1304 00:50:05,600 --> 00:50:06,680 JUST BE NECESSARY FOR 1305 00:50:06,680 --> 00:50:09,200 TRANSCRIPTION TO BE ACTIVATED SO 1306 00:50:09,200 --> 00:50:10,920 BINDING DOESN'T NECESSARILY MEAN 1307 00:50:10,920 --> 00:50:11,640 THAT THERE'S AN EFFECT ON THE 1308 00:50:11,640 --> 00:50:12,720 TRANSCRIPTION SO WE'RE GOING TO 1309 00:50:12,720 --> 00:50:15,480 HAVE TO REALLY CORRELATE THOSE 1310 00:50:15,480 --> 00:50:17,640 WITH OUR DATA ON WHAT GENES 1311 00:50:17,640 --> 00:50:18,720 REQUIRE GLASS FOR THEIR 1312 00:50:18,720 --> 00:50:26,040 EXPRESSION, YEAH, TO BE ABLE TO 1313 00:50:26,040 --> 00:50:26,520 FIGURE THAT OUT. 1314 00:50:26,520 --> 00:50:29,200 >>I'VE BEEN ASKED TO REPEAT THE 1315 00:50:29,200 --> 00:50:33,680 CME CODE, IT'S 444 PIEF 3. 1316 00:50:33,680 --> 00:50:33,960 --44453. 1317 00:50:33,960 --> 00:50:37,320 AND DOCTOR IF YOU HAVE ANY 1318 00:50:37,320 --> 00:50:37,720 CLOSING REMARKS? 1319 00:50:37,720 --> 00:50:38,720 >>WELL IF THERE ARE MOW MORE 1320 00:50:38,720 --> 00:50:41,360 QUESTIONS I WOULD LIKE TO THANK 1321 00:50:41,360 --> 00:50:45,720 JESSICA ONCE MORE FOR A 1322 00:50:45,720 --> 00:50:46,400 FANTASTIC TALK. 1323 00:50:46,400 --> 00:50:49,840 [ APPLAUSE ] 1324 00:50:49,840 --> 00:50:51,440 AND INVITE ALL OF YOU TO STAY 1325 00:50:51,440 --> 00:50:52,640 AROUND A LITTLE BIT FOR MORE 1326 00:50:52,640 --> 00:51:00,640 QUESTIONS AND COFFEE AND COOKIES 1327 00:51:00,640 --> 00:51:01,240 RIGHT OUTSIDE THE AMPHITHEATRE. 1328 00:51:01,240 --> 00:00:00,000 THANK YOU VERY MUCH.