title: Olfactory conditioning in the third instar larvae of Drosophila melanogaster using heat shock reinforcement.
abstract: Adult Drosophila melanogaster has long been a popular model for learning and memory studies. Now the larval stage of the fruit fly is also being used in an increasing number of classical conditioning studies. In this study, we employed heat shock as a novel negative reinforcement for larvae and obtained high learning scores following just one training trial. We demonstrated heat-shock conditioning in both reciprocal and non-reciprocal paradigms and observed that the time window of association for the odor and heat shock reinforcement is on the order of a few minutes. This is slightly wider than the time window for electroshock conditioning reported in previous studies, possibly due to lingering effects of the high temperature. To test the utility of this simplified assay for the identification of new mutations that disrupt learning, we examined flies carrying mutations in the dnc gene. While the sensitivity to heat shock, as tested by writhing, was similar for wild type and dnc homozygotes, dnc mutations strongly diminished learning. We confirmed that the learning defect in dnc flies was indeed due to mutation in the dnc gene using non-complementation analysis. Given that heat shock has not been employed as a reinforcement for larvae in the past, we explored learning as a function of heat shock intensity and found that optimal learning occurred around 41 °C, with higher and lower temperatures both resulting in lower learning scores. In summary, we have developed a very simple, robust paradigm of learning in fruit fly larvae using heat shock reinforcement.
authors.affiliation: Section of Neurobiology and Institute for Neuroscience, University of Texas at Austin, Austin, TX, USA. sukantkhurana@gmail.com
authors.forename: Sukant
authors.initials: S
authors.lastname: Khurana
authors.forename: Brooks G
authors.initials: BG
authors.lastname: Robinson
authors.forename: Zihe
authors.initials: Z
authors.lastname: Wang
authors.forename: William C
authors.initials: WC
authors.lastname: Shropshire
authors.forename: Allen C
authors.initials: AC
authors.lastname: Zhong
authors.forename: Laura E
authors.initials: LE
authors.lastname: Garcia
authors.forename: Jonathan
authors.initials: J
authors.lastname: Corpuz
authors.forename: Jonathan
authors.initials: J
authors.lastname: Chow
authors.forename: Michael M
authors.initials: MM
authors.lastname: Hatch
authors.forename: Eric F
authors.initials: EF
authors.lastname: Precise
authors.forename: Amanda
authors.initials: A
authors.lastname: Cady
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Godinez
authors.forename: Terapat
authors.initials: T
authors.lastname: Pulpanyawong
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Nguyen
authors.forename: Wen-Ke
authors.initials: WK
authors.lastname: Li
authors.forename: Max
authors.initials: M
authors.lastname: Seiter
authors.forename: Kambiz
authors.initials: K
authors.lastname: Jahanian
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Sun
authors.forename: Ruchita
authors.initials: R
authors.lastname: Shah
authors.forename: Sunaina
authors.initials: S
authors.lastname: Rajani
authors.forename: William Y
authors.initials: WY
authors.lastname: Chen
authors.forename: Sofia
authors.initials: S
authors.lastname: Ray
authors.forename: Natalie V
authors.initials: NV
authors.lastname: Ryazanova
authors.forename: Dorah
authors.initials: D
authors.lastname: Wakou
authors.forename: Rohith K
authors.initials: RK
authors.lastname: Prabhu
authors.forename: Nigel S
authors.initials: NS
authors.lastname: Atkinson
mesh_terms: D000818:Animals; D003213:Conditioning, Psychological; D004331:Drosophila melanogaster / Q000196:embryology*; D005816:Genetic Complementation Test; D005824:Genetics, Behavioral / Q000379:methods*; D018869:Heat-Shock Response / Q000235:genetics*; D006358:Hot Temperature; D007814:Larva / Q000235:genetics; D007858:Learning; D008568:Memory; D008957:Models, Genetic; D009154:Mutation; D009812:Odorants; D012054:Reinforcement, Psychology; D012903:Smell / Q000235:genetics*