f learning & memory in flies f environmental sources of plasticity f genetic dissection of...
TRANSCRIPT
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learning & memory in flies environmental sources of plasticity genetic dissection of learning & memory
anatomical mutants biochemical mutants
molecular dissection of memory summary
PART 4: BEHAVIORAL PLASTICITY#23: MOLECULAR GENETICS OF LEARNING
& MEMORY IN DROSOPHILA II
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learning & memory in flies environmental sources of plasticity genetic dissection of learning & memory
anatomical mutants biochemical mutants
molecular dissection of memory summary
PART 4: BEHAVIORAL PLASTICITY#23: MOLECULAR GENETICS OF LEARNING
& MEMORY IN DROSOPHILA II
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PAVLOPAVLOV’S V’S
FLIESFLIES
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2 CLASSES OF L & M MUTANTS
“anatomical” mutants screens for anatomical defects
& “biochemical” mutants
screens for behavioral defects
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2 CLASSES OF L & M MUTANTS
“anatomical” mutants screens for anatomical defects
& “biochemical” mutants
screens for behavioral defects
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BIOCHEMICAL MUTANTS
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MEMORY PHASES
genetically distinct memory phasesJ. Dubnau & T. Tully
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massed trainingrshPKC
cerstau Npum ADF-1 neb CREBCXMspaced trainingadapted from Margulies et al. 2005
... GENETIC DISSECTION
ACQ STM MTM
ARM
LTM
PKA-C1PKA-R1 mbmGs
fas IIleo vollat lio
dnc
rut
amn
PKA-C1
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S. Waddell
MEMORY GENES & SIGNALING
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DROSOPHILA vs APLYSIA
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Crittenden et al. 1998
MEMORY GENE EXPRESSION IN MBs
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one of first transgenic experiments in flies transform flies with
gene for peptide inhibitor of PKA under control of heat-shock promoter... temporal resolution
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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hsp70 XPKA
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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hsp70 XPKA
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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hsp70 XPKA
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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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
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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hsp70 XPKA
permissive temperature: normal learning
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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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
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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hsp70 XPKA
restrictive temperature: learning blocked
cAMP-DEPENDENT PROTEIN KINASE (PKA)
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single cycle training not protein synthesis dependent ARM resistant to cold shock ARM not protein synthesis dependent
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
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long-lasting memory in radish mutants radish encodes some aspect of ARM
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
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transgenic flies: heat-shock activated dominant negative CREB isoform blocks LTM
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
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2 kinds of long-lasting memory massed training ARM, resistant to cold shock spaced training LTM, protein synthesis dependent
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
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transgenic conditional CREB blockers LTM radish mutants have no ARM
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
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GAL4 ENHANCER TRAP SYSTEM
w– E X/
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GAL4 w+
/ w– E X
GAL4 ENHANCER TRAP SYSTEM
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/ GAL4 w+ w– E X
GAL4 ENHANCER TRAP SYSTEM
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/
LacZ w+
w–
GAL4 ENHANCER TRAP SYSTEM
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/ w– LacZ w+
GAL4 ENHANCER TRAP SYSTEM
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/ GAL4 w+ w– E X
/ w– LacZ w+
x
GAL4 ENHANCER TRAP SYSTEM
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/ GAL4 w+ w– E X
/ w– LacZ w+
ß-galactosidase
GAL4 ENHANCER TRAP SYSTEM
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MB-SPECIFIC CELL MARKERS
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G-PROTEINS & cAMP PATHWAY
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/ GAL4 w+ w– E X
/ w– Gs w+
Gs
MB-TARGETED Gs
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Connolly et al.
MB-TARGETED Gs
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ADENYLYL CYCLASE & cAMP PATHWAY
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MB TARGETED rut RESCUE
/ GAL4 w+ w– E X
/ w– rut + w+
adenylate cyclase
rut –
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Zars et al
MB TARGETED rut RESCUE
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/ GAL4 w+ w– E X
/ w– shits w+
dynaminaberrantdynamin
MB NEURON SIGNALING & LEARNING
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Dubnau et al.
MB NEURON SIGNALING & LEARNING
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Dubn
au e
t al.
MB NEURON SIGNALING & RETRIEVAL
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massed trainingrshPKC
cerstau Npum ADF-1 neb CREBCXMspaced trainingadapted from Margulies et al. 2005
... GENETIC DISSECTION
ACQ STM MTM
ARM
LTM
PKA-C1PKA-R1 mbmGs
fas IIleo vollat lio
dnc
rut
amn
PKA-C1
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S. Waddell
MEMORY GENES & SIGNALING
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A.S. Chiang
BUILDING A MEMORY CIRCUIT
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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
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transgenic studies PKA CREB
molecular biology & anatomy: functions defined at cellular level – neurons are equivalent or structural level – neurons are different
SUMMARY
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integrated research approach in fliesCS
CR UR
US
convergence... behavior anatomy cell biology
at all levels of organization... components characterized plasticity
prospects homology applications
SUMMARY