A GAL4(DBD)::Zip- hemidriver (component of the 'Split GAL4' system, contains the GAL4 DNA-binding domain) has been inserted into the first, non-sex specific coding exon of dsx, such that the hemidriver is expressed under the control of endogenous dsx sequences.
Comment: when combined with Mi{Trojan-p65AD.2}Gad1MI09277-Tp65AD.2
Comment: when combined with P{dVP16AD}VGlutOK371-dVP16AD
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with Mi{Trojan-p65AD.2}Gad1MI09277-Tp65AD.2
Comment: when combined with Mi{Trojan-p65AD.2}Gad1MI09277-Tp65AD.2
Comment: when combined with Mi{Trojan-p65AD.2}Gad1MI09277-Tp65AD.2
when combined with P{elav-VP16.AD}
Comment: when combined with P{dVP16AD}VGlutOK371-dVP16AD
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{elav-VP16.AD}
Comment: when combined with P{dVP16AD}VGlutOK371-dVP16AD
when combined with P{elav-VP16.AD}
Comment: when combined with P{dVP16AD}VGlutOK371-dVP16AD
Comment: when combined with P{elav-GAL4.AD}
Scer\GAL4dsx-GAL4-DBD drives expression specifically in neurons when combined with Hsim\VP16AD.elav in a split-GAL4 system including mechanosensory neurons in the terminalia associated with the bristles of the clasper tooth, lateral plates and anal plates in males, and the anal and vaginal plates in females. Scer\GAL4dsx-GAL4-DBD in combination with Hsim\VP16AD-VGlut-OK371 drives expression specifically in a subset of VGlut positive glutamatergic neurons. These include; adult doublesex aDN neurons, motor neurons in the reproductive system and abdomen that arborize onto muscles of the aedeagus, dorsal and central muscles of the sixth abdominal segment, motor neurons that innervate via the abdominal nerve trunk the phallic and periphallic musculature in males and the muscles of the uterus, spermathecal and parovarian ducts in females and prothoracic leg taste bristle chemosensory neurons that project to the prothoracic neuromere and cross the midline in males, but not females. Also included are adult glutamatergic doublesex AbN (male) neurons, adult glutamatergic doublesex AbN (female) neurons and, in males ~8 glutamatergic motor neurons in the abdominal neuromeres that are positive for fru-XP, including those that innervate the muscles of the sixth abdominal segment. Scer\GAL4dsx-GAL4-DBD in combination with Hsap\RELAGad1-MI09277-Tp65AD.2 drives expression specifically in a subset of GABAergic neurons. These include ~1 adult fruitless pIP-e (male) neuron, ~3 adult doublesex SN (male) neurons and, adult GABAergic doublesex AbN (male) neurons and adult GABAergic doublesex AbN (female) neurons. These latter neurons aborize locally, except for two in males that project to the adult mesothoracic and prothoracic neuromeres. pIP-e neurons and adult doublesex SN (male) neurons project within the superior medial protocerebrum and to the adult subesophageal zone. In males, all pIP-e neurons are positive for fru-XP, as is a single adult doublesex SN (male) neuron and ~30 adult GABAergic doublesex AbN (male) neurons.
Blocking synaptic vesicle exocytosis (using Ctet\TeTxLCTNT.Scer\UAS) in the split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsim\VP16AD.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\GAL4dsx-GAL4-DBD and Hsim\VP16AD.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\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-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\TeTxLCScer\FRT.stop.Scer\UAS) in neurons targeted by the split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 along with Scer\FLP1FLPo.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 TrpA1Scer\UAS.(B).cKa at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsim\VP16AD.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\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-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\GAL4dsx-GAL4-DBD/Hsap\RELAGad1-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 TrpA1Scer\FRT.stop.Scer\UAS.T:Hsap\MYC at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 along with Scer\FLP1FLPo.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).