learning & memory in flies environmental sources of plasticity

1. PART 4: BEHAVIORAL PLASTICITY #23: MOLECULAR GENETICS OF LEARNING & MEMORY IN DROSOPHILA II • learning & memory in flies • environmental sources of plasticity • genetic dissection of learning & memory • anatomical mutants • biochemical mutants • molecular dissection of memory • summary

2. PART 4: BEHAVIORAL PLASTICITY #23: MOLECULAR GENETICS OF LEARNING & MEMORY IN DROSOPHILA II • learning & memory in flies • environmental sources of plasticity • genetic dissection of learning & memory • anatomical mutants • biochemical mutants • molecular dissection of memory • summary

3. PAVLOV’S FLIES

4. 2 CLASSES OF L & M MUTANTS • “anatomical” mutants • screens for anatomical defects & • “biochemical” mutants • screens for behavioral defects

5. 2 CLASSES OF L & M MUTANTS • “anatomical” mutants • screens for anatomical defects & • “biochemical” mutants • screens for behavioral defects

6. BIOCHEMICAL MUTANTS

7. MEMORY PHASES • genetically distinct memory phases J. Dubnau & T. Tully

8. ARM   LTM ... GENETIC DISSECTION massed training rsh PKC cer stau N pum ADF-1 neb CREB CXM spaced training PKA-C1 PKA-R1 mbm Gs fas II leo vol lat lio dnc rut amn PKA-C1  ACQ STM  MTM adapted from Margulies et al. 2005

9. MEMORY GENES & SIGNALING S. Waddell

10. DROSOPHILA vs APLYSIA

11. MEMORY GENE EXPRESSION  IN MBs Crittenden et al. 1998

12. cAMP-DEPENDENT PROTEIN KINASE (PKA) • one of first transgenic experiments in flies • transform flies with • gene for peptide inhibitor of PKA • under control of heat-shock promoter... • temporal resolution

13. hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

15. hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

16. hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

17. cAMP-DEPENDENT PROTEIN KINASE (PKA) • one of first transgenic experiments in flies • transform flies with • gene for peptide inhibitor of PKA • under control of heat-shock promoter... • temporal resolution • test associative learning • permissive temperature: normal learning

18. hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)  • permissive temperature: normal learning

19. cAMP-DEPENDENT PROTEIN KINASE (PKA) • one of first transgenic experiments in flies • transform flies with • gene for peptide inhibitor of PKA • under control of heat-shock promoter... • temporal resolution • test associative learning • permissive temperature: normal learning • restrictive temperature: blocked learning

20. hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA) • restrictive temperature: learning blocked 

21. CREB cAMP-RESPONSE ELEMENT BINDING PROTEIN • single cycle training not protein synthesis dependent • ARM resistant to cold shock • ARM not protein synthesis dependent

22. CREB cAMP-RESPONSE ELEMENT BINDING PROTEIN • long-lasting memory in radish mutants • radish encodes some aspect of ARM

23. CREB cAMP-RESPONSE ELEMENT BINDING PROTEIN • transgenic flies: heat-shock activated dominant negative CREB isoform blocks LTM

24. CREB cAMP-RESPONSE ELEMENT BINDING PROTEIN • 2 kinds of long-lasting memory • massed training  ARM, resistant to cold shock • spaced training  LTM, protein synthesis dependent

25. CREB cAMP-RESPONSE ELEMENT BINDING PROTEIN • transgenic conditional CREB blockers LTM • radish mutants have no ARM

26. / w– E X GAL4 ENHANCER TRAP SYSTEM

28. GAL4 w+ / w– E X GAL4 ENHANCER TRAP SYSTEM

29. GAL4 w+ / w– E X GAL4 ENHANCER TRAP SYSTEM

31. LacZ w+ GAL4 ENHANCER TRAP SYSTEM / w–

32. LacZ w+ / w– GAL4 ENHANCER TRAP SYSTEM

33. GAL4 w+ LacZ w+ GAL4 ENHANCER TRAP SYSTEM /   w– E X x / w–

34. GAL4 w+ LacZ w+ ß-galactosidase GAL4 ENHANCER TRAP SYSTEM / w– E X / w–

35. MB-SPECIFIC CELL MARKERS

36. G-PROTEINS & cAMP PATHWAY

37. GAL4 w+ Gs w+ Gs MB-TARGETED Gs / w– E X / w–

38. MB-TARGETED Gs Connolly et al.

39. ADENYLYL CYCLASE & cAMP PATHWAY

40. GAL4 w+ rut + w+ adenylate cyclase MB TARGETED rut RESCUE / w– E X / w– rut –

41. MB TARGETED rut RESCUE Zars et al

42. GAL4 w+ shits w+ aberrant dynamin dynamin MB NEURON SIGNALING & LEARNING / w– E X / w–

43. MB NEURON SIGNALING & LEARNING Dubnau et al.

44. MB NEURON SIGNALING & RETRIEVAL Dubnau et al.

45. ARM   LTM ... GENETIC DISSECTION massed training rsh PKC cer stau N pum ADF-1 neb CREB CXM spaced training PKA-C1 PKA-R1 mbm Gs fas II leo vol lat lio dnc rut amn PKA-C1  ACQ STM  MTM adapted from Margulies et al. 2005

46. MEMORY GENES & SIGNALING S. Waddell

47. BUILDING A MEMORY CIRCUIT A.S. Chiang

48. SUMMARY • flies can learn a variety of tasks • classical differential conditioning of odor avoidance • genetic dissection: mutants impaired in • cAMP cascade • mushroom body neuroanatomy • ... all have correlated learning / memory defects • homology with other species / model systems • memory phases • single cycle training • massed & spaced training

49. SUMMARY • transgenic studies • PKA • CREB • molecular biology & anatomy: functions defined at • cellular level – neurons are equivalent • or • structural level – neurons are different

50. CS CR UR US    SUMMARY • integrated research approach in flies • convergence... • behavior • anatomy • cell biology • at all levels of organization... • components characterized • plasticity • prospects • homology • applications