Integration of Parallel Opposing Memories Underlies Memory Extinction

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yes a) No extinction. 7 na b) 2 CS-, 15 min ITI. 7 na c) 2 CS+, 15 min ITI. 7 na a) R58E02-GAL4 control CS- exposed. 8 yes b) R58E02-GAL4 control CS+ ...
Cell, Volume 175

Supplemental Information

Integration of Parallel Opposing Memories Underlies Memory Extinction Johannes Felsenberg, Pedro F. Jacob, Thomas Walker, Oliver Barnstedt, Amelia J. Edmondson-Stait, Markus W. Pleijzier, Nils Otto, Philipp Schlegel, Nadiya Sharifi, Emmanuel Perisse, Carlas S. Smith, J. Scott Lauritzen, Marta Costa, Gregory S.X.E. Jefferis, Davi D. Bock, and Scott Waddell

Table  1.  Statistical  details  related  to  Figure  1-­6  and  S1-­S7 Experiment  /  Figure

n

normally  distributed

Figure  1A

a)  No  extinction b)  2  CS+,  1  min  ITI c)  2  CS+,  15  min  ITI d)  5  CS+,  1  min  ITI

group/treatment

6 10 10 10

na yes yes yes

statistical  test

ANOVA  =  0.0025,  Turkey's:  a  vs  b  =   ANOVA  with  Tukey's  multiple  comparisons   0.99,  a  vs  c  =  0.02,  a  vs  d  =  0.04,  b  vs  c   test =  0.01,  b  vs  d  =  0.03,  c  vs  d  =  0.98

p  value

Figure  1B

a)  No  extinction b)  2  CS-­,  15  min  ITI c)  2  CS+,  15  min  ITI

7 7 7

na na na

ANOVA  with  Tukey's  multiple  comparisons   test

Figure  1C

a)  R58E02-­GAL4  control  CS-­  exposed b)  R58E02-­GAL4  control  CS+  exposed c)  UAS-­shits  control  CS-­  exposed d)  UAS-­shits  control  CS+  exposed e)  R58E02-­GAL4;;  UAS-­Shits1    CS-­  exposed f)  R58E02-­GAL4;;  UAS-­Shits1    CS+  exposed

8 7 8 7 8 7

yes na yes na yes na

ANOVA  (CS-­)  =  0.0081,  Turkey's:  a  vs   unpaired  t-­test  for  comparision  between   c  =  0.0292,  a  vs  e  =  0.0104,  c  vs  e  =   same  genotype  but  different  treatment,   0.8768,  ANOVA  (CS+)  =  0.0046,   ANOVA  with  Tukey's  multiple  comparisons   Turkey's:  b  vs  d  =  0.88,  b  vs  f  =  0.02,  d   test  for  comparision  between  different   vs  f  =  0.006,  unpaired  t-­test:  a  vs  b  <   genotype,  same  treatment 0.0001,  c  vs  d  =  0.02,  e  vs  f  =  0.4298

Figure  1  D

a)  MB504B-­GAL4  control  CS-­  exposed b)  MB504B-­GAL4  control  CS+  exposed c)  UAS-­Shits  control  CS-­  exposed d)  UAS-­Shits  control  CS+  exposed e)  MB504B-­GAL4;;  UAS-­Shits1    CS-­  exposed f)  MB504B-­GAL4;;  UAS-­Shits1    CS+  exposed

6 6 6 6 6 6

na na na na na na

unpaired  t-­test  for  comparision  between   same  genotype  but  different  treatment,   ANOVA  (CS-­)  =  0.8581,  ANOVA  (CS+)   ANOVA  with  Tukey's  multiple  comparisons   =  0.9763,  unpaired  t-­test:  a  vs  b  =   test  for  comparision  between  different   0.001,  c  vs  d  =  0.01,  e  vs  f  =  0.03 genotype,  same  treatment

g  5 g  5  control g4 b'2p b'2m

5 4 5 4 4

na na na na na

a)  VT1211-­GAL4  control  CS+  exposed b)  UAS-­Shits1  control  CS+  exposed c)  VT1211-­GAL4;;  UAS-­Shits1    CS+  exposed a)  VT1211-­GAL4  control  CS-­  exposed c)  UAS-­Shits1  control  CS-­  exposed e)  VT1211-­GAL4;;  UAS-­Shits1    CS-­  exposed

16 16 16 9 9 9

yes no yes yes yes yes

Figure  3A Figure  3B Figure  3C Figure  3D Figure  3E Figure  3F

MB112C-­GAL4  dendrites  training  no  extinction  MCH  as  CS+ R39A05-­GAL4  dendrites  training  no  extinction  MCH  as  CS+ R66C08-­GAL4  dendrites  training  no  extinction  MCH  as  CS+ MB112C-­GAL4  dendrites  training  extinction  MCH  as  CS+ R39A05-­GAL4  dendrites  training  extinction  MCH  as  CS+ R66C08-­GAL4  dendrites  training  extinction  MCH  as  CS+

11 12 14 15 10 12

Figure  6A Figure  6B Figure  6C Figure  6D Figure  6E Figure  6F

VT1211-­GAL4  axons  no  extinction  MCH  as  CS+ R39A05-­GAL4  axons  training  no  extinction  MCH  as  CS+ R66C08-­GAL4  axons  training  no  extinction  MCH  as  CS+ VT1211-­GAL4  axons  training  extinction  MCH  as  CS+ R39A05-­GAL4  axons  training  extinction  MCH  as  CS+ R66C08-­GAL4  axons  training  extinction  MCH  as  CS+

Figure  S1A

ANOVA  =  0.0025,  Turkey's:  a  vs  b  =   0.97,  a  vs  c  =  0.003,  b  vs  c  =  0.004

paired  t-­test paired  t-­test paired  t-­test paired  t-­test paired  t-­test

p  =  0.0086 p  =  0.2664 p  =  0.2059 p  =  0.061 p  =  0.1805

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  =  0.0108,  Turkey's:  a  vs  b  =   0.99,  a  vs  e  =  0.02,  b  vs  c  =  0.02

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  <  0.0001,  Turkey's:  a  vs  b  =   0.88,  a  vs  e  <  0.0001,  b  vs  c  =  0.0003

no no no no yes yes

Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test paired  t-­test

p=0.0186 p=0.8501 p=0.6698 p=0.0125 p=0.3506 p=0.0019

15 14 18 14 16 11

no yes yes no no yes

Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test

p=0.0151 p=0.0378 p=0.0299 p=0.2166 p=0.0250 p=0.2594

 a)  2  pre-­exposures  1  min  ITI,  10-­3    b)  2  pre-­exposures  15  min  ITI,  10-­3    c)  15  pre-­exposures  1  min  ITI,  10-­3    d)  2  pre-­exposures  15  min  ITI,  10-­4  a)  2  pre-­exposures  1  min  ITI,  10-­6  b)  2  pre-­exposures  15  min  ITI,  10-­6  c)  5  pre-­exposures  1  min  ITI,  10-­6

7 8 8 8 11 11 11

na yes yes yes yes yes yes

One  sample  t  test  (theoretical  mean  =  0),   ANOVA  with  Tukey's  multiple  comparisons   test

t-­test:    p  <  0.0001,  ANOVA  <  0.0001,   Turkey's:  a  vs  b  <  0.0001,  a  vs  c  <   0.0001,  b  vs  c  =  0.246

10-­3 10-­6

6 6

na na

a)  training/extinction/retraining/immediate  test b)  training/-­-­-­/retraining/immediate  test c)  -­-­-­/-­-­-­/retraining/immediate  test

9 8 9

yes yes yes

a)  training/extinction/retraining/  90  min  test b)  training/-­-­-­/retraining/  90  min  test c)  -­-­-­/-­-­-­/retraining/  90  min  test

9 9 9

yes yes yes

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  =  0.0137,  Turkey's:  a  vs  b  =   0.975,  a  vs  c  =  0.02,  b  vs  c  =  0.03

Figure  S1D

a)  R58E02-­GAL4  control  CS-­  exposed b)  R58E02-­GAL4  control  CS+  exposed c)  UAS-­Shits1  control  CS-­  exposed d)  UAS-­Shits1  control  CS+  exposed e)  R58E02-­GAL4;;  UAS-­Shits1    CS-­  exposed f)  R58E02-­GAL4;;  UAS-­Shits1    CS+  exposed

6 6 6 6 6 6

na na na na na na

unpaired  t-­test  for  comparision  between   same  genotype  but  different  treatment,   ANOVA  (CS-­)  =  0.1770,  ANOVA  (CS+)   ANOVA  with  Tukey's  multiple  comparisons   =  0.9490,  unpaired  t-­test:    a  vs  b  =   test  for  comparision  between  different   0.0003,  c  vs  d  =  0.0012,  e  vs  f  =  0.02 genotype,  same  treatment

Figure  S1E

a)  R58E02-­GAL4  CS+  exposed c)  UAS-­shits  CS+  exposed e)  R58E02-­GAL4;;  UAS-­Shits1  CS+  exposed

9 9 9

yes yes yes

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  =  0.411

Figure  S2A

a)  VT1211-­GAL4   c)  UAS-­Shits1   b)  VT1211-­GAL4

9 9 9

yes yes yes

ANOVA  with  Tukey's  multiple  comparisons   test  for  comparision

ANOVA  =  0.3359

a)  VT1211-­GAL4  control  CS-­  exposed c)  UAS-­Shits1  control  CS-­  exposed b)  VT1211-­GAL4  control  CS-­  exposed

11 11 10

yes yes yes

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  =  0.5785

d)  VT1211-­GAL4  control  CS+  exposed e)  UAS-­Shits1  control  CS+  exposed f)  VT1211-­GAL4  control  CS+  exposed

11 11 10

yes no yes

ANOVA  with  Tukey's  multiple  comparisons   test  for  comparision

ANOVA  =  0.6085

Figure  S3A Figure  S3B Figure  S3C Figure  S3D Figure  S3E Figure  S3F Figure  S3G Figure  S3H Figure  S3I Figure  S3J Figure  S3K Figure  S3L Figure  S3M Figure  S3N Figure  S3O Figure  S3P Figure  S3Q Figure  3R

R66C08-­GAL4  dendrites  training  no  extinction  OCT  as  CS+ R66C08-­GAL4  dendrites  training  extinction  OCT  as  CS+ MB112C-­GAL4  dendrites  training  no  extinction  OCT  as  CS+ MB112C-­GAL4  dendrites  training  extinction  OCT  as  CS+ VT1211-­GAL4  dendrites  training  no  extinction  MCH  as  CS+ VT1211-­GAL4  dendrites  training  extinction  MCH  as  CS+ VT1211-­GAL4  dendrites  training  no  extinction  OCT  as  CS+ VT1211-­GAL4  dendrites  training  extinction  OCT  as  CS+ R39A05-­GAL4  dendrites  training  no  extinction  OCT  as  CS+ R39A05-­GAL4  dendrites  training  extinction  OCT  as  CS+ MB112C-­GAL4  dendrites  mock  no  extinction  OCT  as  CS+ MB112C-­GAL4  dendrites  mock  extinction  OCT  as  CS+ MB112C-­GAL4  dendrites  mock  no  extinction  MCH  as  CS+ MB112C-­GAL4  dendrites  mock  extinction  MCH  as  CS+ VT1211-­GAL4  dendrites  mock  no  extinction  MCH  as  CS+ VT1211-­GAL4  dendrites  mock  extinction  MCH  as  CS+ VT1211-­GAL4  dendrites  mock  no  extinction  OCT  as  CS+ VT1211-­GAL4  dendrites  mock  extinction  OCT  as  CS+

11 13 17 15 14 13 14 11 16 12 11 12 18 15 11 11 11 10

yes yes yes no no no yes yes yes yes no no no no no no yes no

paired  t-­test paired  t-­test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test paired  t-­test paired  t-­test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test

p=0.9359 p=0.0417 p=0.0107 p=0.0302 p=0.3258 p=0.0215 p=0.3803 p=0.0104 p=0.1284 p=0.0934 p>0.999 p=0.7334 p=0.7987 p=0.9341 p=0.7002 P=0.8984 p=0.8764 p=0.7695

Figure  S6A Figure  S6B Figure  S6C Figure  S6D Figure  S6E Figure  S6F Figure  S6G Figure  S6H Figure  S6I Figure  S6J

VT1211-­GAL4  axons  training  no  extinction  OCT  as  CS+ VT1211-­GAL4  axons  training  extinction  OCT  as  CS+ R66C08-­GAL4  axons  training  no  extinction  OCT  as  CS+ R66C08-­GAL4  axons  training  extinction  OCT  as  CS+ R39A05-­GAL4  axons  training  no  extinction  OCT  as  CS+ R39A05-­GAL4  axons  training  extinction  OCT  as  CS+ VT1211-­GAL4  axons  mock  no  extinction  MCH  as  CS+ VT1211-­GAL4  axons  mock  extinction  MCH  as  CS+ VT1211-­GAL4  axons  mock  no  extinction  OCT  as  CS+ VT1211-­GAL4  axons  mock  no  extinction  OCT  as  CS+

11 10 24 12 14 16 12 11 10 11

yes yes no no yes no no yes no yes

paired  t-­test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test Wilcoxon  matched-­pairs  signed  rank  test paired  t-­test

p=0.0065 p=0.7695 p=0.0457 p=0.9697 p=0.0378 p=0.0250 p=0.1099 p=0.2848 p=0.7695 p=0.1923

Figure  2  A  and  B Figure  2  C Figure  2  D Figure  2  E

Figure  2  F

Figure  S1B

Figure  S1C  left

Figure  S1C  right

Figure  S2B

One  sample  t  test  (theoretical  mean  =  0)

p  =  0.843

One  sample  t  test  (theoretical  mean  =  0)

t-­test:  a  =  0.883,  b  =  0.290,  c  =  0.22,   ANOVA:  p  =  0.675

paired  t-­test

p  =  0.4852

ANOVA  with  Tukey's  multiple  comparisons   test

ANOVA  =0.7637