Thermogenetic activation of P-Fr neurons via expression of TrpA1^UAS.(B).cKa under the control of Scer\GAL4^GMR37G12 or P-Fr split-Gal4 (Hsap\RELA^AD.R37G12 with Scer\GAL4^{DBD.VT036267.PX}) leads to significantly less sleep at night time, a lower arousal threshold at night time (only with Scer\GAL4^GMR37G12), and a significant increase in locomotion of flies during daytime (with Scer\GAL4^GMR37G12 but not with with P-Fr split-Gal4) and nighttime (with P-Fr split-Gal4 but not with Scer\GAL4^GMR37G12) when compared to control adults.

Expression of TrpA1^UAS.(B).cKa under the control of Scer\GAL4^TrpA1-AB-G4 in larvae results in a shift of larval rolling (escape) behaviors and heat-induced locomotor arrest towards occurring at lower temperatures, compared to controls, with reduced levels of rolling overall.

Thermogenetic activation of neurons labelled by Scer\GAL4^Tk (either the P{Tk-GAL4.TH}5Fa or P{Tk-GAL4.TH}3Ma insertion) through expression of TrpA1^{Scer\UAS.(B).cKa} at 6 days after eclosion strongly increases male aggression compared to controls. No increase in aggression is seen when TrpA1^{Scer\UAS.(B).cKa} is expressed under the control of another insertion of the same construct, P{Tk-GAL4.TH}2Ma. Activation of P{Tk-GAL4.TH}5Fa neurons also increase aggression towards a wild-type male.

No increase in male aggression is seen compared to controls when neurons labelled by Scer\GAL4^GMR61H07 are activated by inducing expression of TrpA1^{Scer\UAS.(B).cKa} at 6 days after eclosion by shifting flies to 29[o].

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of any one of Scer\GAL4^Kurs58, Scer\GAL4^c767, Scer\GAL4^OK307, Scer\GAL4^dimm-929, Scer\GAL4^Aph-4-c507, Scer\GAL4^53b, Scer\GAL4^NP5221, Scer\GAL4^ey-OK107, Scer\GAL4¹⁴⁷¹, Scer\GAL4^c320 or Scer\GAL4^GMR65C11 at 28[o]C (which results in activation of the warmth-activated TrpA1^{Scer\UAS.(B).cKa} protein) results in a degradation of circadian locomotor activity rhythms under constant darkness conditions compared to controls kept at 21[o]C. When flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Kurs58 are returned to 21[o]C, locomotor activity rhythms are recovered at a similar phase as control lines.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Ilp2.PR at 28[o]C has no effect on locomotor activity rhythms under constant darkness conditions compared to controls kept at 21[o]C.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^qvr.PW (using a temperature pulse of 27[o]C from ZT18-ZT24 to activate TrpA1) results in a reduction in sleep relative to controls.

Activation of GABA+ neurons through heat-induced expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Gad1.PS inhibits baseline female contact-induced male-male aggression.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^tank-4-12 at 30[o]C increases ethanol sensitivity in males (time required for 50% of flies to sedate upon exposure to 60% ethanol is reduced compared to controls). No significant effect on ethanol sensitivity is seen in females expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^tank-4-12 at 30[o]C.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^9-30 or Scer\GAL4^11-81 at 27[o]C increases ethanol sensitivity in males (time required for 50% of flies to sedate upon exposure to 60% ethanol is reduced compared to controls). No significant effect on ethanol sensitivity is seen in females expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^9-30 or Scer\GAL4^11-81 at 27[o]C.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^{natalisin.Hsim\VP16} at 30[o]C show significantly reduced mating frequency compared to wild-type males.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^E564 at 32[o]C suppresses the proboscis extension response over a range of sucrose concentrations when the stimulation is applied to the leg, while the proboscis extension response to sucrose is unaffected if the stimulation is applied to the proboscis.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Crz.PU during adulthood (flies raised at 18[o]c than shifted to 29[o]C for behavioural testing) results in a significant increase in ethanol sedation sensitivity compared to wild type.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^elav-C155 or Scer\GAL4^{Akap200-NP2222} triggers seizure-like output from the central pattern generator in larvae in a temperature-dependent manner, with increased motor neuron bursting at 27[o]C and continuous seizure activity at 30[o]C.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^elav-C155 or Scer\GAL4^{Akap200-NP2222} results in rapid-onset seizures in adult flies upon exposure to 30[o]C.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^repo results in paralysis at 38[o]C.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NP6510 (flies raised at 31[o]C for 12 hours and 25[o]C for 12 hours each day) show a progressive decline in locomotor performance in a startle-induced negative geotaxis (SING) assay.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^c305a and incubated at 31[o]C for 10 minutes prior to testing show a significant reduction in locomotor performance in a SING assay compared to unheated controls.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Mef2.247 and incubated at 31[o]C for 10 minutes prior to testing do not show a significant difference in locomotor performance in a SING assay compared to unheated controls.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^futsch-C380 at 29[o]C results in severe dendritic defects in the MN5 motorneurons if the flies are shifted to 29[o]C between pupal stages P2 and P6 or between P2 and P9. Dendritic defects are not seen if the temperature shift is during stages P6 to P9.

Thermo-activation of dopamine neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^ple.PF for 60 seconds at 30[o]C during odor presentation induces immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies. Co-expressing Scer\GAL80^Cha.3.3P does not suppress this response.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Ddc.PL for 60 seconds at 30[o]C during odor presentation does not induce the immediate aversive memory (two minutes after odor presentation) seen in wild type flies in response to punishment. A similar lack of aversive memory is seen when TrpA1^{Scer\UAS.(B).cKa} is expressed under the control of Scer\GAL4^Ddc.HL9.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} and Scer\GAL80^Mef2.mb247 under the control of Scer\GAL4^NP0047 for 60 seconds at 30[o]C during odor presentation induces immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies. Co-expression of Scer\GAL80^ple.PK suppresses this response. In contrast, co-expression of Scer\GAL4^NP5272 is unable to significantly alter immediate memory retention at 2 minutes, but improves memory retention two hours after odor presentation. This enhancement is suppressed when the flies are exposed to cold shock after training (to remove anesthesia resistant memory). Aversive memory can also be induced by expressing TrpA1^{Scer\UAS.(B).cKa} in flies that are maintained at the permissive temperature and exposed to electric shock during odor presentation. The extent of the memory induced by this method is identical regardless of whether it is expressed under the control of Scer\GAL4^NP0047 alone or in conjunction with Scer\GAL4^NP5272.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^c259 and Scer\GAL80^Mef2.mb247 for 60 seconds at 30[o]C during odor presentation induces immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies. Co-expression of Scer\GAL80^ple.PK partially suppresses this response.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^5-HT1B.PA for 60 seconds at 30[o]C during odor presentation induces a slight but significant immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies. Co-expression of Scer\GAL80^ple.PK or Scer\GAL80^Cha.3.3P suppresses this response. The extent of the memory performance is dependent to some extent on the activation temperature, with some memory formation also seen at both 28[o]C and 29[o]C. Although the memory performance induced using Scer\GAL4^5-HT1B.PA is moderate, it remains stable at this level up to nine hours after odor presentation. Co-expression of Scer\GAL4^NP5272 does not significantly enhance immediate memory retention, but improves memory retention two hours after odor presentation. This enhancement is suppressed when the flies are exposed to cold shock after odor presentation (to remove anesthesia resistant memory). Aversive memory can also be induced by expressing TrpA1^{Scer\UAS.(B).cKa} in flies that are maintained at the permissive temperature and exposed to electric shock during odor presentation. The extent of the memory induced by this method is identical regardless of whether it is expressed under the control of Scer\GAL4^5-HT1B.PA alone or in conjunction with Scer\GAL4^NP5272.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NP7187 for 60 seconds at 30[o]C during odor presentation does not induce the immediate aversive memory (two minutes after odor presentation) seen in wild type flies in response to punishment.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^{CG17646-NP7323} for 60 seconds at 30[o]C during odor presentation slight but significant amount of immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Mz19 and Scer\GAL80^Mef2.mb247 for 60 seconds at 30[o]C during odor presentation does not induce the immediate aversive memory (two minutes after odor presentation) seen in wild type flies in response to punishment.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^c061 and Scer\GAL80^Mef2.mb247 for 60 seconds at 30[o]C during odor presentation induces the immediate aversive memory (two minutes after odor presentation) seen in response to punishment in wild type flies. The extent of the memory performance is dependent on the activation temperature, as flies are able to form significant memory at 27[o]C-29[o]C, but to a lesser extent than at 30[o]C. Whilst the memory response is initially robust, it decays rapidly over nine hours. The more moderate memory induced when expression is induced at 27[o]C disappears completely within three hours. Co-expression of Scer\GAL4^NP5272 does not significantly enhance memory retention at any time points tested (two minutes, two hours and nine hours) and exposure to cold shock after training (to remove anesthesia resistant memory) also has no effect. Co-expression of Scer\GAL4^5-HT1B.PA is also unable to suppress aversive memory at 30[o]C, but memory impairment can be induced at 28[o]C, as is seen when TrpA1^{Scer\UAS.(B).cKa} is driven by Scer\GAL4^5-HT1B.PA alone. Aversive memory can also be induced by expressing TrpA1^{Scer\UAS.(B).cKa} in flies that are maintained at the permissive temperature and exposed to electric shock during odor presentation. The extent of the memory induced by this method is identical regardless of whether it is expressed under the control of Scer\GAL4^c061 alone or in conjunction with either Scer\GAL4^c061 and Scer\GAL4^5-HT1B.PA.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NP5272 for 60 seconds at 30[o]C during odor presentation induces a small amount of immediate aversive memory (two minutes after odor presentation), as is seen in response to punishment in wild type flies. No memory formation is seen at lower activation temperatures. Whilst the memory response at 30[o]C is robust immediately after odor presentation it decays completely within 3 hours.

Thermo-activation of neurons by expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Mz840 and Scer\GAL80^Mef2.mb247 for 60 seconds at 30[o]C during odor presentation does not produce the immediate aversive memory (two minutes after odor presentation) seen in wild type flies in response to punishment. Co-expression of Scer\GAL4^NP5272 is unable to induce memory formation (at two hours after odor presentation).

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC at 31[o]C contingent with odour presentation is sufficient to form appetitive olfactory memory in satiated flies. Significant memory persists at 30 minutes after training in satiated flies but does not remain at 3 hours, even in hungry flies.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^NP7088, Scer\GAL4⁰⁶⁶⁵ or Scer\GAL4⁰⁸⁹¹ at 31[o]C contingent with odour presentation does not induce the formation of appetitive olfactory memory.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4⁰¹⁰⁴ or Scer\GAL4⁰²⁷³ at 31[o]C contingent with odour presentation is sufficient to form appetitive olfactory memory.

Flies carrying TrpA1^{Scer\UAS.(B).cKa} and Scer\GAL4^AstA.2.1 which are raised at 22[o]C and then shifted to 28[o]C an hour before testing show a reduction in the fraction of flies that feed within 1 hour following 24 hours starvation compared to control flies.

Expression of TrpA1^{Scer\UAS.(B).cKa} at the permissive temperature (27[o]C) under the control of Scer\GAL4^ple.PF increases the proportion of female flies that show a sugar-sensitive proboscis extension reflex (PER). This phenotype is observed within 10 minutes of the temperature shift.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Gr5a.8.5 confers sensitivity to N-methylmaleimide in L-type labellar taste bristles (robust spiking is seen in a in an extracellular tip recording assay in response to 10mM N-methylmaleimide). The L-type taste bristles are thermosensitive in these animals.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Gr66a.PD results in 75% of flies regurgitating liquid upon being warmed to 32[o]C after being allowed to ingest water to satiation.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^ple.PF which have been shifted to 30[o]C for 45 minutes and subsequently returned to 25[o]C show markedly reduced courtship towards mated females compared to that of control males. Courtship towards virgin females is normal.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Ddc.HL9 which have been shifted to 30[o]C for 45 minutes and subsequently returned to 25[o]C show markedly reduced courtship towards mated females compared to that of control males.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^ple.PF, Scer\GAL4^ple.TH-D' or Scer\GAL4^ple.TH-D at 27[o]C results in a strong reduction in nighttime sleep compared to controls, with a decrease in sleep bout duration and sleep bout number. These sleep defects are suppressed by expression of Scer\GAL80^ple.PK.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^ple.TH-D at 27[o]C show a significant reduction in arousal threshold compared to controls when exposed to graded light pulses at zeitgeber time 16.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^ple.TH-D-4 or Scer\GAL4^ple.TH-G-1 at 27[o]C results in a significant reduction in nighttime sleep and sleep bout duration compared to controls. Night-time sleep bout number and daytime sleep are unaffected. The sleep defects are suppressed by expression of Scer\GAL80^ple.PK.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC at 27[o]C results in a moderate reduction in nighttime sleep compared to controls, with a decrease in sleep bout duration, not no affect on sleep bout number.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^n-syb.PS at 27[o]C results in a strong reduction in nighttime sleep compared to controls. This sleep defect is largely suppressed by Scer\GAL80^ple.PK.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^l.Trh.PK, Scer\GAL4^ple.TH-F-1, Scer\GAL4^ple.TH-F-2 or Scer\GAL4^ple.TH-F-3 at 27[o]C has no significant effect on nighttime sleep.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^upd-E132 at 28[o]C results in defects in locomotor activity rhythms (only 35.7% of flies show rhythmic behaviour under constant darkness conditions). Locomotor activity rhythms revert back to normal after the flies are shifted to 21[o]C.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B, Scer\GAL4^fru.P1.D, Scer\GAL4^dsx-GAL4 or Scer\GAL4^dsx.Δ2 at 27[o]C show substantial courtship towards freely-moving females in a vision-based courtship assay (females separated from male targets by a thin transparent plastic barrier). This contrasts with wild-type males which barely court females under these conditions. The mutant males show low levels of courtship towards immobile females under these conditions.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^GMR71G01 at 30[o]C readily court freely-moving females but not immobile females in a vision-based courtship assay. These males also court constantly rotating rubber band pieces of approximately the size of a fly in this assay.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NPF.1 at 29[o]C results in an aversion to ethanol-supplemented food in virgin males.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NPF.1 and trained (in the presence of an odour) for three 1 hour sessions at 29[o]C, interspersed with three 1-hour rest periods at 18[o]C (in the absence of the odour) show a preference for the odour when tested at 21[o]C 24 hours after training.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^NPF.1 at 30[o]C abolishes conditioned preference for ethanol in virgin males when the males are tested 24 hours after training.

Thirty-minute exposure of Scer\GAL4^GH146>TrpA1^{Scer\UAS.(B).cKa} flies to 29[o]C is sufficient to induce a reduction in the avoidance response to 5% ethyl butyrate but not to CO[[2]]. In contrast, similar activation in V projection neurons (under the control of Scer\GAL4^VPN) results in reduced olfactory avoidance to CO[[2]] but has no effect on the ethyl butyrate response. Similar to odorant-induced habituation, the projection-induced reduced olfactory avoidance response recovers substantially after 1 hour at room temperature. In addition, this learned reduced avoidance can be erased by brief, intense mechanical stimulation or by exposure to strong novel odorants.

Copulation duration is reduced in males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Trh.1.0 and shifted to the activating temperature of 28-31[o]C approximately 60 seconds after the start of mating.

Copulation duration is increased in males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Trh.1.0 and shifted to the activating temperature of 28-31[o]C approximately 10 minutes after the start of mating.

Tethered males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Crz.391 and shifted to the activating temperature of 28-31[o]C ejaculate within approximately 60 seconds of the temperature shift. This phenotype is also seen in males which are decapitated before being shifted to the activating temperature.

Tethered males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Crz.2fa and shifted to the activating temperature of 28-31[o]C do not ejaculate.

Tethered males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^Trh.1.0 or Scer\GAL4^CrzR.3.5 and shifted to the activating temperature of 28-31[o]C ejaculate.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^Trh.1.0, Scer\GAL4^Crz.391 or Scer\GAL4^CrzR.3.5 in males and shifting to the activating temperature during copulation (1 minute after the start of mating) results in precocious sperm and seminal fluid transfer to females. Copulation also terminates prematurely, after only approximately 7 minutes, when expression is driven by Scer\GAL4^CrzR.3.5.

At a permissive temperature, Scer\GAL4^ppk23.2.695-driven expression of TrpA1^{Scer\UAS.(B).cKa} males court oenocyte-ablated males and females, as expected (ablation by a Scer\GAL4^desat1.PB, Scer\GAL80^ts.αTub84B/W^Scer\UAS.cZa background). When TrpA1^{Scer\UAS.(B).cKa} is activated at 30[o]C, these flies exhibit reduced courtship towards oe[-] males and increased courtship towards oe[-] females. When paired with oe[+] flies (containing pheromones) at 30[o]C, these flies do not court oe[+] flies but increase courtship towards oe[+] females.

Males in which expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^ppk23.2.336 is activated by increased temperature show courtship towards wild-type males.

Larvae expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^ppk.1.9 have a dramatically lowered thermal nociception threshold. Greater than 90% of these larvae respond to a 30[o]C heat stimulus with nocifensive escape locomotion in less than 1 second. No response is seen in controls.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC at 29[o]C significantly reduces male courtship toward females. If expression is activated in males during training with virgin females courtship conditioning is artificially induced in the male flies.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Oamb.KI at 29[o]C does not reduce male courtship toward females.

Adults expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Lk.2.2 during the adult stage have a higher defecation rate and produce less concentrated fecal deposits than control flies.

Overexpression of TrpA1^{Scer\UAS.(B).cKa} in PDF neurons under the control of Scer\GAL4^P2.4.Pdf results in an arrhythmic behavioral rhythm phenotype, but no change in the levels of fat body triglycerides.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B or Scer\GAL4^fru.P1.D at 29[o]C in solitary males rapidly elicits almost all steps of male courtship behaviour in every male analysed. The wild-type courtship behaviours observed include unilateral wing extension and vibration, proboscis extension, abdomen bending, attempted copulation and ejaculation. In addition, bilateral wing extension and vibration is elicited in these flies, a behaviour that is not normally characteristic of wild-type courtship. An array of courtship behaviours and bilateral wing extension is still induced in solitary males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B or Scer\GAL4^fru.P1.D at 29[o]C even when the flies are decapitated.

Solitary males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^fru.P1.D at 27[o]C initially show unilateral wing extension, which transitions to bilateral wing extension. The transition occurs more quickly in decapitated males.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^dsx-GAL4 or Scer\GAL4^dsx.Δ2 at 29[o]C in solitary males rapidly elicits almost all steps of male courtship behaviour in every male analysed. The wild-type courtship behaviours observed include unilateral wing extension and vibration, proboscis extension, abdomen bending and attempted copulation. Ejaculation is not seen in a 15 minute observation period, but a few flies ejaculate in a longer observation period of 30 minutes. Bilateral wing extension and vibration is elicited in these flies, a behaviour that is not normally characteristic of wild-type courtship. An array of courtship behaviours and bilateral wing extension is still induced in solitary males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^dsx-GAL4 or Scer\GAL4^dsx.Δ2 at 29[o]C even when the flies are decapitated.

Solitary males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^dsx.Δ2 at 27[o]C show consistent unilateral wing extension (and not bilateral wing extension) through a 1 hour observation period. However, if the males are decapitated, they show exclusively bilateral wing extension under these conditions.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B or Scer\GAL4^fru.P1.D at 29[o]C is sufficient to induce unilateral wing extension in females, although they produce atypical songs compared to male songs.

The normal order of behaviours is seen in the courtship behaviours elicited in intact solitary males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B, Scer\GAL4^fru.P1.D, Scer\GAL4^dsx-GAL4 or Scer\GAL4^dsx.Δ2 at 27 or 29[o]C, in that wing extension is always induced before abdomen bending. This ordering is lost in headless males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of any one of these drivers.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^fru.P1.B or Scer\GAL4^fru.P1.D at 27[o]C display intense levels of courtship towards D. melanogaster females (both intact and headless) and copulate with intact females at frequencies similar to wild-type controls. The mutant males also court headless D. melanogaster females and headless females of other species more intensely than do control males.

Males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of either Scer\GAL4^dsx-GAL4 or Scer\GAL4^dsx.Δ2 at 27[o]C vigorously court both intact and headless D. melanogaster females, but do not show significantly higher courtship of headless females of other species than at the permissive temperature of 22[o]C.

Expression of TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^l.Trh.PK results in enhancement of serotonergic neuron function. Pairing of males of these flies results in mid-intensity aggression with 90% of these pairings resulting in the establishment of dominance relationships. This is not statistically different from control flies. The latency to establish dominance, the number of lunges before winning and the low numbers of lunges displayed by 'loser' flies are no different to control flies. However, the latency to the first lunge is significantly shortened and the number of lunges performed by 'winners' after establishing dominance are dramatically increased.

The total courtship activity of male flies expressing TrpA1^{Scer\UAS.(B).cKa} driven by Scer\GAL4^Tdc2.PC is not significantly altered compared to controls. Although courtship initiation by Scer\GAL4^Tdc2.PC-driven TrpA1^{Scer\UAS.(B).cKa} expressing males towards females is more rapid than controls, successful copulation rates are not significantly different. Like control males, males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC also spend more time singing to females than to males.

The percentage of time in which Scer\GAL4^Tdc2.PC-driven TrpA1^{Scer\UAS.(B).cKa} expressing males court other males rather than females is significantly increased compared with controls.

When courted by other males, wild-type males show rejection behaviour, including vigorous wing flicking directed at the courter. This and other sensory cues usually lead to a rapid termination of all aspects of male-male courtship behaviour in wild-type males. In males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC, unlike in wild-type males, the average duration of wing extension in male-male and male-female singing bouts is essentially identical and at levels normally seen during male-female courtship behaviour.

In contrast to wild-type males, courtship patterns interrupt aggression displays in males expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^Tdc2.PC.

Expression of TrpA1^{Scer\UAS.(B).cKa} in Scer\GAL4^ple.PF-expressing cells at 15[o]C, when TrpA1 is inactive, results in low levels of locomotor activity. When the temperature is raised to 30[o]C, above the temperature required for TrpA1 activation, Scer\GAL4^ple.PF expression of TrpA1^{Scer\UAS.(B).cKa} increases locomotor activity levels. Returning these flies to 15[o]C almost immediately results in a cessation of motion. Moreover, repeated temperature cycling causes repeated cycling between states of inactivity and activity.

Activation of TrpA1^{Scer\UAS.(B).cKa} in Scer\GAL4^c346 expressing neurons results in increased locomotor activity.

Expression of TrpA1^{Scer\UAS.(B).cKa} in Scer\GAL4^Ddc.HL5 expressing neurons has no effect on locomotor activity.

In Scer\GAL4^c346; Scer\GAL80^ple.PK/+; TrpA1^{Scer\UAS.(B).cKa}/+ flies (where Scer\GAL80^ple.PK blocks expression), no stimulation of locomotor activity is detected.

Food deprived and trained Scer\GAL4^NPF.1; TrpA1^{Scer\UAS.(B).cKa} flies, fed ad libitum for 3 hours, show robust appetitive memory at the permissive temperature of 23[o]C, as in wild-type. However, stimulation of NPF neurons for 30 minutes before and during testing by shifting the flies to 31[o]C reveals memory performance in Scer\GAL4^NPF.1;TrpA1^{Scer\UAS.(B).cKa} flies that is statistically different from controls.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} in MB-MP neurons (mushroom body innervating DA neurons), under the control of Scer\GAL4^c061 and Scer\GAL80^Mef2.mb247 display robust appetitive memory at 23[o]C. MB-MP neuron stimulation of these flies (through a temperature shift to 31[o]C) prior to and during memory retrieval severely impairs memory. Food-deprived Scer\GAL4^c061; Scer\GAL80^Mef2.mb247/TrpA1^{Scer\UAS.(B).cKa} flies stimulated during acquisition or for 1 hour after training show normal 3 hour memory.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} in MB-MP neurons (mushroom body innervating DA neurons), under the control of Scer\GAL4^{kra-krasavietz} and Scer\GAL80^Mef2.mb247 display robust appetitive memory at 23[o]C. MB-MP neuron stimulation of these flies (through a temperature shift to 31[o]C) prior to and during memory retrieval severely impairs memory.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} in MB-MP neurons (mushroom body innervating DA neurons), under the control of Scer\GAL4^IP3K2-NP2758 display robust appetitive memory at 23[o]C. MB-MP neuron stimulation of these flies (through a temperature shift to 31[o]C) prior to and during memory retrieval severely impairs memory.

MNISN-Is neurons expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^futsch-C380 fire tonically in response to modest heat pulses.

Larvae expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^VGlut-OK371 are quickly immobilised by increase surface temperature to 27[o]C. Crawling speeds start to recover as the surface temperature gradually falls back to 22[o]C.

Larvae expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^5-40 are almost completely immobile at 27[o]C. As the surface temperature than falls, normal crawling speeds do not resume until 50 seconds after the surface temperature has returned to 22[o]C.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^elav-C155 are incapacitated after 60 seconds at 35[o]C (this phenotype is not seen in controls), but they recover on return to 23[o]C.

Flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^elav-C155 show abnormal electrophysiology in response to moderate warming; above 25[o}C, a barrage of excitatory junctional potentials are triggered at the neuromuscular junction.

Overexpression of TrpA1^{Scer\UAS.(B).cKa} with Scer\GAL4^dimm-929 results in a decrease in sleep during the nighttime compared to controls when entrained for 5 days in LD at 25[o]C, switched to 30[o]C for 2 days and then back to 25[o]C. Males of the genotype TrpA1^{Scer\UAS.(B).cKa};Scer\GAL4^dimm-929;Scer\GAL80^Pdf.PS sleep significantly less than controls immediately after temperature elevation from 25[o]C to 30[o]C.

Heat-induced activation of TrpA1 at ZT12 in flies expressing TrpA1^{Scer\UAS.(B).cKa} under the control of Scer\GAL4^dimm-929 induces increased locomotor activity.

Scer\GAL4^Tk, TrpA1^UAS.(B).cKa has abnormal behavior | adult stage | heat sensitive phenotype, enhanceable by Tk^UAS.cAa/Scer\GAL4^Tk

Scer\GAL4^GH146, TrpA1^UAS.(B).cKa has abnormal smell perception | heat sensitive phenotype, enhanceable by Nmdar1^RNAi.UAS.cWa, Scer\GAL4^GH146

Scer\GAL4^VPN, TrpA1^UAS.(B).cKa has abnormal smell perception | heat sensitive phenotype, enhanceable by Nmdar1^RNAi.UAS.cWa, Scer\GAL4^VPN

Scer\GAL4^GH146, TrpA1^UAS.(B).cKa has abnormal smell perception | heat sensitive phenotype, enhanceable by Rdl^{RNAi.8-10.UAS}, Scer\GAL4^GH146

Scer\GAL4^VPN, TrpA1^UAS.(B).cKa has abnormal smell perception | heat sensitive phenotype, enhanceable by Rdl^{RNAi.8-10.UAS}, Scer\GAL4^VPN

Scer\GAL4^VPN, TrpA1^UAS.(B).cKa has abnormal smell perception | heat sensitive phenotype, enhanceable by shakB²

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has abnormal neuroanatomy | adult stage phenotype, non-enhanceable by mir-263b^Δ/mir-263b^Δ

Scer\GAL4^NPF.PU, TrpA1^UAS.(B).cKa has abnormal feeding behavior | larval stage | heat sensitive phenotype, non-enhanceable by Dop1R1^KK102341, Scer\GAL4^NPF.PU

Scer\GAL4^GMR58E02, TrpA1^UAS.(B).cKa has abnormal memory | adult stage | heat sensitive phenotype, non-enhanceable by Tbh^nM18

Scer\GAL4^ppk.1.9, TrpA1^UAS.(B).cKa has abnormal pain response | larval stage phenotype, suppressible | partially by Dsk^2A-GAL4, TrpA1^UAS.(B).cKa

Scer\GAL4^AstC-2A-GAL4, Scer\GAL80^R57C10, TrpA1^UAS.(B).cKa has increased rate of feeding behavior phenotype, suppressible by AstC^UAS.KO.pCFD6/Spyo\Cas9^UAS.P2, Scer\GAL4^AstC-2A-GAL4, Scer\GAL80^R57C10

Scer\GAL4^VP16.CNMa, TrpA1^UAS.(B).cKa has decreased fecundity | female phenotype, suppressible by AstC¹/AstC¹

Scer\GAL4^Ptth.cMa, TrpA1^UAS.(B).cKa has abnormal developmental rate | larval stage phenotype, suppressible by Ptth^120F2A

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has abnormal feeding behavior | larval stage | heat sensitive phenotype, suppressible | partially by Hug^RNAi.UAS.1A, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has abnormal neurophysiology | larval stage | heat sensitive phenotype, suppressible by Hug^RNAi.UAS.1A, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has abnormal feeding behavior | larval stage | heat sensitive phenotype, suppressible | partially by ChAT^JF01877, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has abnormal neurophysiology | larval stage | heat sensitive phenotype, suppressible by ChAT^JF01877, Scer\GAL4^S3.Hug

Scer\GAL4^GMR50H05, TrpA1^UAS.(B).cKa has abnormal feeding behavior | adult stage phenotype, suppressible | partially by Trhn^RNAi.1.UAS/Scer\GAL4^GMR50H05

Scer\GAL4^GMR50H05, TrpA1^UAS.(B).cKa has abnormal feeding behavior | adult stage phenotype, suppressible | partially by Scer\GAL4^GMR50H05/Trhn^RNAi.2.UAS

Scer\GAL4^Tk, TrpA1^UAS.(B).cKa has abnormal behavior | adult stage | heat sensitive phenotype, suppressible | partially by Df(3R)ΔTk²/Tk^Δ1

Scer\GAL4^Tk, TrpA1^UAS.(B).cKa has abnormal behavior | adult stage | heat sensitive phenotype, suppressible by TkR86C^ΔF28

Scer\GAL4^repo, TrpA1^UAS.(B).cKa has paralytic | adult stage | heat sensitive phenotype, suppressible by Cam^HMS01318, Scer\GAL4^repo

Scer\GAL4^ple.PF, TrpA1^UAS.(B).cKa has abnormal courtship behavior | heat sensitive phenotype, suppressible by Or67d^GAL4-1

Scer\GAL4^ple.PF, TrpA1^UAS.(B).cKa has abnormal sleep | heat sensitive phenotype, suppressible by Dop1R1^f02676/Dop1R1^f02676

Scer\GAL4^AstC.PM, TrpA1^UAS.(B).cKa has abnormal oviposition phenotype, non-suppressible by AstC¹/AstC¹

Scer\GAL4^AstC.PM, TrpA1^UAS.(B).cKa has decreased fecundity | female phenotype, non-suppressible by AstC¹/AstC¹

Scer\GAL4^VP16.CNMa, TrpA1^UAS.(B).cKa has decreased fecundity | female phenotype, non-suppressible by CNMa¹/CNMa¹

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has abnormal neuroanatomy | adult stage phenotype, non-suppressible by mir-263b^Δ/mir-263b^Δ

Scer\GAL4^NPF.PU, TrpA1^UAS.(B).cKa has abnormal feeding behavior | larval stage | heat sensitive phenotype, non-suppressible by Dop1R1^KK102341, Scer\GAL4^NPF.PU

Scer\GAL4^Tk, TrpA1^UAS.(B).cKa has abnormal behavior | adult stage | heat sensitive phenotype, non-suppressible by TkR99D^MB09356

Scer\GAL4^ple.PF, TrpA1^UAS.(B).cKa has abnormal sleep | heat sensitive phenotype, non-suppressible by Dop1R2^MB05108/Dop1R2^MB05108

Scer\GAL4^ple.PF, TrpA1^UAS.(B).cKa has abnormal sleep | heat sensitive phenotype, non-suppressible by Dop2R^f06521

Scer\GAL4^ple.PF, TrpA1^UAS.(B).cKa has abnormal sleep | heat sensitive phenotype, non-suppressible by DopEcR^c02142/DopEcR^c02142

Scer\GAL4^GMR58E02, TrpA1^UAS.(B).cKa has abnormal memory | adult stage | heat sensitive phenotype, non-suppressible by Tbh^nM18

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of abnormal neuroanatomy | adult stage phenotype of Pdf⁰¹

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of abnormal circadian rhythm | adult stage phenotype of Pdf⁰¹

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of abnormal neuroanatomy | adult stage phenotype of Pdfr⁵³⁰⁴

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of abnormal circadian rhythm | adult stage phenotype of Pdfr⁵³⁰⁴

Dsk^2A-GAL4, TrpA1^UAS.(B).cKa is a suppressor | partially of abnormal pain response | larval stage phenotype of Scer\GAL4^ppk.1.9, TrpA1^UAS.(B).cKa

Scer\GAL4^Piezo.1.0/TrpA1^UAS.(B).cKa is a suppressor of increased rate of feeding behavior | recessive | adult stage phenotype of Piezo^KO

Scer\GAL4^Piezo.1.0/TrpA1^UAS.(B).cKa is a suppressor of increased size | recessive | adult stage phenotype of Piezo^KO

Scer\GAL4^Ptth.cMa/TrpA1^UAS.(B).cKa is a suppressor of abnormal developmental rate | larval stage phenotype of Ptth^120F2A

Scer\GAL4^Gr32a.3.776/TrpA1^UAS.(B).cKa is a suppressor of abnormal courtship behavior | male phenotype of Gr32a^unspecified

Scer\GAL4^Gr28b.d.PT/TrpA1^UAS.(B).cKa is a suppressor | partially of abnormal temperature response phenotype of Gr28b^MB03888

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of abnormal neuroanatomy | adult stage phenotype of Pdf⁰¹

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of abnormal circadian rhythm | adult stage phenotype of Pdf⁰¹

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of abnormal neuroanatomy | adult stage phenotype of Pdfr⁵³⁰⁴

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of abnormal circadian rhythm | adult stage phenotype of Pdfr⁵³⁰⁴

TrpA1^UAS.(B).cKa/Scer\GAL4^GH146 is a non-suppressor of abnormal smell perception | heat sensitive phenotype of shakB²

Scer\FLP1^{FLPo.oc.Tag:NLS(SV40-largeT)}, Scer\GAL4^dsx.KI, Scer\GAL80^αTub84B.PG, TrpA1^UAS.(B).cKa, dsx^TH13362.N has abnormal mating behavior phenotype

Hsap\RELA^AD.R37G12, Scer\GAL4^{DBD.VT036267.PX}, TrpA1^UAS.(B).cKa has abnormal locomotor behavior | adult stage | heat sensitive phenotype

Hsap\RELA^AD.R37G12, Scer\GAL4^{DBD.VT036267.PX}, TrpA1^UAS.(B).cKa has abnormal sleep | adult stage | heat sensitive phenotype

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has axon collateral | adult stage phenotype, non-enhanceable by mir-263b^Δ/mir-263b^Δ

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has s-LNv Pdf neuron phenotype, non-enhanceable by mir-263b^Δ/mir-263b^Δ

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has pharyngeal muscle cell | heat sensitive phenotype, suppressible by Hug^RNAi.UAS.1A, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has embryonic/larval antennal nerve | heat sensitive phenotype, suppressible by Hug^RNAi.UAS.1A, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has pharyngeal muscle cell | heat sensitive phenotype, suppressible by ChAT^JF01877, Scer\GAL4^S3.Hug

Scer\GAL4^S3.Hug, TrpA1^UAS.(B).cKa has embryonic/larval antennal nerve | heat sensitive phenotype, suppressible by ChAT^JF01877, Scer\GAL4^S3.Hug

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has s-LNv Pdf neuron phenotype, non-suppressible by mir-263b^Δ/mir-263b^Δ

Scer\GAL4^Pdf.PU, TrpA1^UAS.(B).cKa has axon collateral | adult stage phenotype, non-suppressible by mir-263b^Δ/mir-263b^Δ

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of adult s-LNv neuron | adult stage phenotype of Pdf⁰¹

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-enhancer of adult s-LNv neuron | adult stage phenotype of Pdfr⁵³⁰⁴

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of adult s-LNv neuron | adult stage phenotype of Pdfr⁵³⁰⁴

TrpA1^UAS.(B).cKa/Scer\GAL4^Pdf.Switch1a is a non-suppressor of adult s-LNv neuron | adult stage phenotype of Pdf⁰¹