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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
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
PAVLOPAVLOV’S V’S
FLIESFLIES
2 CLASSES OF L & M MUTANTS
“anatomical” mutants screens for anatomical defects
& “biochemical” mutants
screens for behavioral defects
2 CLASSES OF L & M MUTANTS
“anatomical” mutants screens for anatomical defects
& “biochemical” mutants
screens for behavioral defects
BIOCHEMICAL MUTANTS
MEMORY PHASES
genetically distinct memory phasesJ. Dubnau & T. Tully
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
S. Waddell
MEMORY GENES & SIGNALING
DROSOPHILA vs APLYSIA
Crittenden et al. 1998
MEMORY GENE EXPRESSION IN MBs
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)
hsp70 XPKA
cAMP-DEPENDENT PROTEIN KINASE (PKA)
hsp70 XPKA
cAMP-DEPENDENT PROTEIN KINASE (PKA)
hsp70 XPKA
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
cAMP-DEPENDENT PROTEIN KINASE (PKA)
hsp70 XPKA
permissive temperature: normal learning
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
cAMP-DEPENDENT PROTEIN KINASE (PKA)
hsp70 XPKA
restrictive temperature: learning blocked
cAMP-DEPENDENT PROTEIN KINASE (PKA)
single cycle training not protein synthesis dependent ARM resistant to cold shock ARM not protein synthesis dependent
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
long-lasting memory in radish mutants radish encodes some aspect of ARM
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
transgenic flies: heat-shock activated dominant negative CREB isoform blocks LTM
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
2 kinds of long-lasting memory massed training ARM, resistant to cold shock spaced training LTM, protein synthesis dependent
CREBcAMP-RESPONSE ELEMENT BINDING PROTEIN
transgenic conditional CREB blockers LTM radish mutants have no ARM
CREBcAMP-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–
GAL4 ENHANCER TRAP SYSTEM
/ w– LacZ w+
GAL4 ENHANCER TRAP SYSTEM
/ GAL4 w+ w– E X
/ w– LacZ w+
x
GAL4 ENHANCER TRAP SYSTEM
/ GAL4 w+ w– E X
/ w– LacZ w+
ß-galactosidase
GAL4 ENHANCER TRAP SYSTEM
MB-SPECIFIC CELL MARKERS
G-PROTEINS & cAMP PATHWAY
/ GAL4 w+ w– E X
/ w– Gs w+
Gs
MB-TARGETED Gs
Connolly et al.
MB-TARGETED Gs
ADENYLYL CYCLASE & cAMP PATHWAY
MB TARGETED rut RESCUE
/ GAL4 w+ w– E X
/ w– rut + w+
adenylate cyclase
rut –
Zars et al
MB TARGETED rut RESCUE
/ GAL4 w+ w– E X
/ w– shits w+
dynaminaberrantdynamin
MB NEURON SIGNALING & LEARNING
Dubnau et al.
MB NEURON SIGNALING & LEARNING
Dubn
au e
t al.
MB NEURON SIGNALING & RETRIEVAL
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
S. Waddell
MEMORY GENES & SIGNALING
A.S. Chiang
BUILDING A MEMORY CIRCUIT
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
transgenic studies PKA CREB
molecular biology & anatomy: functions defined at cellular level – neurons are equivalent or structural level – neurons are different
SUMMARY
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
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