Blocking synaptic vesicle exocytosis (using Ctet\TeTxLC^TNT.Scer\UAS) in the split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsim\VP16^{AD.elav.T:Zzzz\Zip+,T:SV40\nls-largeT} neurons in males and females results in a significant drop (compared to genetic controls) in courtship of females by males, males completely fail to copulate and are behaviorally sterile; females are sterile, unreceptive and show no post-mating behavioral responses. Blocking synaptic vesicle exocytosis in neurons targeted by the combination of Scer\GAL4^dsx-GAL4-DBD and Hsim\VP16^{AD.VGlut.OK371.T:Zzzz\Zip+,T:SV40\nls-largeT} results in a significant drop in male courtship behavior (they display the normal complement of courtship behavior including attempts to copulate but spend less time courting target females), fertility (they produce no progeny), and no successful matings occur (males fail to achieve genital coupling) compared to genetic controls. Blocking synaptic vesicle exocytosis in neurons targeted by the split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsap\RELA^{Gad1-MI09277-Tp65AD.2} does not significantly affect male courtship behavior (they display the normal complement of courtship behaviors including attempts to copulate and relatively normal levels of courtship toward target females), but less than 20% of males successfully copulate (during a 1hr period) and those that do copulate take significantly longer than controls and 80% could not uncouple from the female ('stuck' phenotype); male fertility is also significantly reduced compared to genetic controls. Blocking synaptic vesicle exocytosis (using Ctet\TeTxLC^{Scer\FRT.stop.Scer\UAS}) in neurons targeted by the split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsap\RELA^{Gad1-MI09277-Tp65AD.2} along with Scer\FLP1^{FLPo.oc.Tag:NLS(SV40-largeT)} results in a significant reduction of male courtship behavior but no copulation defects (successful matings in 1hr and copulation duration are not significantly different compared to genetic controls).

Thermogenetic activation (using TrpA1^{Scer\UAS.(B).cKa} at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsim\VP16^{AD.VGlut.OK371.T:Zzzz\Zip+,T:SV40\nls-largeT} results in a significant drop in male courtship behavior (they display the normal complement of courtship behaviors but spend less time courting target females and never successfully copulate), and no successful matings occur compared to genetic or temperature controls. Thermogenetic activation of these neurons 5 minutes into copulation (when fertilization is not complete) prevents the male from detaching from the female and terminating copulation; decapitation of these males in copulo does not eliminate the 'stuck' phenotype. Thermogenetic activation of neurons targeted by the Split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsap\RELA^{Gad1-MI09277-Tp65AD.2} does not significantly affect male courtship behavior, but completely blocks genital attachment (no successful matings) in the 1hr observation period. 90% of Scer\GAL4^dsx-GAL4-DBD/Hsap\RELA^{Gad1-MI09277-Tp65AD.2} males dismount females and terminate copulation within one minute of thermogenetic activation in copulo; matings truncated in this way never produce progeny. Thermogenetic activation (using TrpA1^{Scer\FRT.stop.Scer\UAS.T:Hsap\MYC} at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4^dsx-GAL4-DBD and Hsap\RELA^{Gad1-MI09277-Tp65AD.2} along with Scer\FLP1^{FLPo.oc.Tag:NLS(SV40-largeT)} (both pre-copulation and in copulo) does not significantly impair male courtship (time spent in courtship behavior is similar to genetic controls) or copulation (successful matings in 1hr and copulation duration is similar to genetic controls).