Identification and localization of the Protein Kinase C requirement for operant self-learning in Drosophila
Julien Colomb
1, Björn Brembs
21 FU Berlin, Institut für Biologie - Neurobiologie, Königin-Luise-Strasse 28/30, 14195 Berlin, Germany.
2 Universität Regensburg, Institute of Zoology - Neurogenetics, Universitätsstrasse 31, 93040 Regensburg, Germany
http://lab.brembs.net
SfN meeting 2012 New Orleans
582.23/QQ9
Putative mutants
Putative mutants RNAi using TARGET RNAi using TARGET
−1.0
−0.5 0.0 0.5 1.0
U-PKC53e_RNAi CS
U-PKC53e_RNAielavG4,tubG80ts U-PKCInac_RNAielavG4,tubG80ts
learning Index
Wtb Pinac PKC53e PKCdelta
−1.0−0.50.00.51.0learning Index
finding a working heat shock protocol:
tub-Gal80ts;UAS-PKCi x elav-Gal4 lines
finding a working heat shock protocol:
tub-Gal80ts;UAS-PKCi x elav-Gal4 lines
Candidate screen Candidate screen motorneuron line
tub-Gal80ts;UAS-PKCi x c380-Gal4, chaGal80
motorneuron line
tub-Gal80ts;UAS-PKCi x c380-Gal4, chaGal80
* *
gen. controls MB,
central complex
d42-Gal4 pan-neuronalglutamatergic neurons
n>25
CS
c380G4,chaG80
32
11 13
CS
elavGal4
U-PKCi, tGal80ts CS
U-PKCi, tGal80tselavGal4
36° heatshock, 1-2h, 34° 4Hh
17 15 32
−1
−0.5 0 0.5 1
35.5° heatshock , 4h
6 10 11
CS
elavGal4
U-PKCi, tGal80ts CS
U-PKCi, tGal80tselavGal4
learning Index
−1
−0.5 0 0.5 1
learning Index
−1
−0.5 0 0.5 1
U-PKCi, tGal80ts CS
learning Index
−1
−0.5 0 0.5 1
U-PKCi, tGal80tsc380G4,chaG80
MB: H24
central complex: c232 and 007y;c819 pan-neuronal: elav glutamatergic: OK371
19°
>33°
enhancer tubulin UAS
promoter
UAS Gal 4
PKCi/RNAi Gal80 ts
PKCi/RNAi
-Outlook
Conclusions Conclusions
Using an inhibitory peptide of Protein Kinase C (PKCi) we have been able to discover a novel learning mechanism underlying operant self-learning. This mechanism appears to be conserved in the bilaterian clade and also involves the Drosophila orthologue of the human 'language gene' FOXP2, dFoxP. Experiments using the TARGET system to temporally restrict RNAi expression to adulthood yielded PKC53e as the putative PKC isoform involved in self-learning. Spatially restricting PKCi expression with the TARGET system revealed that PKC activity is required in glutamatergic neurons. Data are currently ambiguous if motorneurons are the responsible glutamatergic subpopulation and further, potentially disambiguating studies are currently ongoing.
Introduction Introduction
−1.5
−1.0
−0.5 0.0 0.5 1.0 1.5
Self-learning: control flies, n=104
training test training training
pretest training test
lamp
shutter
laser
light guide & arena camera
Did language evolve using an
ancestral self-learning mechanism?
dFoxP-dependent dFoxP-dependent
Colomb J, Brembs B. The biology of psychology: “Simple” conditioning? Communicative & integrative biology. 2010;3(2):142-5
operant behavior Behavior
“US”
world learning self
learning
PKC cAMP
“CS”
dependentMB
Goal directed Behavior Crystallized
Behavior
Habit formation
over-training
world- and self-learning interact during operant conditioning
torque meter
yaw torque signal
diffusor light guides light source
IR laser diode
solenoid
color filter
Control Behavior
heat color
world l.
torque meter
yaw torque signal
diffusor light guides light source
IR laser diode
Control Behavior
heat
self l.
PKC-dependent
adenylate cyclase type I dependentself l.
8 min 8 min
torque meter
yaw torque signal
diffusor light guides light source
IR laser diode
solenoid
color filter
Control Behavior
heat color
world l.
self l.
16 min
FoxP1 FoxP2 FoxP3 FoxP4 FoxP
ancestral Fox P
Urbilaterian (>530 Mya)
Vertebr ates
Invertebr ates
Vocalizations Babble modifies
cause
Vocalizations Subsong modifies
cause
Balancing Falling
modifies causes
Turning Heat
modifies causes
