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

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

PAVLOV’S FLIES

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

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

BIOCHEMICAL MUTANTS

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

massed training rsh PKC cer stauN pumADF-1 nebCREB CXM spaced training adapted from Margulies et al GENETIC DISSECTION  ACQ  STM  MTM   ARM LTM PKA-C1 PKA-R1 mbm G  s fas II leovol latlio dnc rut amn PKA-C1

S. Waddell MEMORY GENES & SIGNALING

DROSOPHILA vs APLYSIA

Crittenden et al MEMORY GENE EXPRESSION  IN MBs

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

hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

hsp70 XPKA cAMP-DEPENDENT PROTEIN KINASE (PKA)

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

hsp70 XPKA E permissive temperature: normal learning cAMP-DEPENDENT PROTEIN KINASE (PKA)

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

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

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

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

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

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

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

GAL4 ENHANCER TRAP SYSTEM w – E X /

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

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

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

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

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

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

MB-SPECIFIC CELL MARKERS

G-PROTEINS & cAMP PATHWAY

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

Connolly et al. MB-TARGETED G  s

ADENYLYL CYCLASE & cAMP PATHWAY

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

Zars et al MB TARGETED rut RESCUE

/ GAL4 w + w – E X / w– w– shi ts w + dynamin aberrant dynamin MB NEURON SIGNALING & LEARNING

Dubnau et al. MB NEURON SIGNALING & LEARNING

Dubnau et al. MB NEURON SIGNALING & RETRIEVAL

massed training rsh PKC cer stauN pumADF-1 nebCREB CXM spaced training adapted from Margulies et al GENETIC DISSECTION  ACQ  STM  MTM   ARM LTM PKA-C1 PKA-R1 mbm G  s fas II leovol latlio dnc rut amn PKA-C1

S. Waddell MEMORY GENES & SIGNALING

A.S. Chiang BUILDING A MEMORY CIRCUIT

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

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

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