Co-expression of WScer\UAS.cZa and rprScer\UAS.cZa under the control of Scer\GAL4ptc-559.1 is lethal, but is viable when Scer\GAL80αTub84B.PL is expressed until the third instar larval stage. Robust cell death is seen in the wing disc 12 hours after Scer\GAL4 de-repression.
When larvae expressing WScer\UAS.cZa and rprScer\UAS.cZa under the control of Scer\GAL4ptc-559.1 are irradiated (and expression is repressed until 12 hours prior to irradiation using Scer\GAL80αTub84B.PL), less cell death is seen in the wing disc in comparison with irradiated controls. A heterozygous Df(3L)banΔ1 background has no effect on the amount of cell death, but when de-repression occurs 6 hours prior to irradiation (which results in a less dramatic reduction in cell death compared to 12 hours), heterozygous Df(3L)banΔ1 partially suppresses the level of protection seen when WScer\UAS.cZa and rprScer\UAS.cZa are expressed alone.
When wing discs containing WScer\UAS.cZa and rprScer\UAS.cZa expressing clones (repressed during early development using Scer\GAL80αTub84B.PL)) are irradiated, less cell death is seen outside of the clones in comparison with irradiated controls.
A Ras85DR68Q background can partially suppress the hemocyte death seen upon expression of WScer\UAS.cZa under the control of Scer\GAL4Hml.PG. Hemocytes are clearly visible anteriorly in the lymph glands of 3rd instar larvae. Circulating hemocytes, however, appear to remain susceptible to W-induced cell death and are missing, even in the presence of two copies of Ras85DR68Q.
Ablation of male oenocytes (and associated cuticular hydrocarbons) through expression of WScer\UAS.cZa, and Avic\GFPStinger.Scer\UAS.T:nls-tra under the control of Scer\GAL4desat1.PB and Scer\GAL80ts.αTub84B at 18[o]C during development and 25[o]C at the adult stage results in substantially reduced levels of male-male aggression compared with pairs of control males, in addition to higher levels of male-male courtship. However, aggression between such oenocyte-eliminated males is not completely eliminated. Control flies show markedly lower levels of male-male aggression towards oenocyte-eliminated males than control males, as measured by the occurrence of lunges. Wing threat display is unaffected by the elimination of male cuticular hydrocarbons. Control males show higher levels of courtship towards oenocyte-eliminated than toward control target males, as measured by the occurrence of unilateral wing extensions and circling episodes. Restoring cuticular hydrocarbons to oenocyte-eliminated males through passive transfer from control flies rescues both the decrease male-male aggression and increase male-male courtship.
Passive addition of synthetic (z)-7-tricosene to oenocyte-eliminated male flies (through expression of WScer\UAS.cZa, and Avic\GFPStinger.Scer\UAS.T:nls-tra under the control of Scer\GAL4desat1.PB and Scer\GAL80ts.αTub84B at 18[o]C during development and 25[o]C at the adult stage) restores normal levels of male-male aggression, and suppresses the increased levelsl of male-male courtship, by wild-type males.
Males co-expressing WScer\UAS.cZa and rprScer\UAS.C under the control of Scer\GAL4CheB42a.ppk25 show courtship latency (time from female introduction until the male shows any courtship related behaviours) similar to that of parental controls.
Males co-expressing WScer\UAS.cZa and rprScer\UAS.C under the control of Scer\GAL4CheB42a.ppk25 show a mild change in courtship index (the proportion of time a male spends courting once courting started) which is not statistically significant compared to controls.
Scer\GAL4srp.D.cCa-mediated expression of rprScer\UAS.cZa and WScer\UAS.cZa leads to a high lethality rate during early larval development - these larvae are devoid of hemocytes and rarely reach the third instar larval stage.
Expression of both rprScer\UAS.cZa and WScer\UAS.cZa in the cortex and neuropile glia from late embryonic stage onwards, under the control of Scer\GAL4nrv2.PS at 18[o]C and switching to a high temperature (25[o]C) shortly after hatching allows most animals to develop to late larval stages. In these larvae, glial cell death sets in around hatching and is more or less complete by the early 2nd instar. These larval brains exhibit a greatly increased number of intra-neuropilar tracheal branches. Six or more secondary branches split off the perineuropilar plexus at variable positions and penetrate the neuropile. Larvae lacking cortex and neuropile glia (but retaining surface glia) show severe behavioural deficits, characterised by reduced peristaltic movement and feeding, and a constant 'tremor' of the pharyngeal and body wall musculature.
Males co-expressing rprScer\UAS.cZa and WScer\UAS.cZa under the control of Scer\GAL4IFa.PT show no difference in male-female courtship behaviour compared to control males when naive males are allowed to court virgin wild-type females. However, the mutant males show significant male-male courtship behaviour when naive males are allowed to court males.
Animals co-expressing rprScer\UAS.cZa and WScer\UAS.cZa under the control of Scer\GAL4P2.4.Pdf are arrhythmic for locomotor activity under constant light conditions at a constant temperature of 25[o]C, but become rhythmic almost immediately after being exposed to a temperature cycle of 12 hours at 25[o]C and 12 hours at 30[o]C. There is a peak at the beginning of the cryophase and a smaller peak at the early thermophase. The flies show a dispersed anticipatory activity starting around the middle of the thermophase, although transient cycles are not clear.
The electroantennogram (EAG) rhythm in response to ethyl acetate (measured on the second day of constant darkness conditions) in flies co-expressing rprScer\UAS.cZa and WScer\UAS.cZa under the control of Scer\GAL4P2.4.Pdf is similar to that of wild-type flies.
Third instar larvae co-expressing both rprScer\UAS.cZa and WScer\UAS.cZa under the control of Scer\GAL41118 have no obvious defects in the Bolwig nerve axonal projections although the lateral neurons are completely missing.
Co-expression of WScer\UAS.cZa and rprScer\UAS.cZa (when driven by Scer\GAL4Gp150-52A) results in ectopic midline cell death. The addition of thScer\UAS.T:Ivir\HA1 suppresses this phenotype, sometimes to wild-type.
When grimScer\UAS.cNa and rprScer\UAS.cZa are coexpressed, all the midline glia and some VUM neurons are eliminated. When grimScer\UAS.cNa and WScer\UAS.cZa are coexpressed, all the midline glia and VUM neurons are eliminated.
Mediated expression of both rprScer\UAS.cZa and WScer\UAS.cZa causes a striking loss of glial cells, significant defects in axon guidance scaffold are detected. VUM neurons exhibit greatly reduced sensitivity: cells are normal in position, number and morphology. Two copies of each construct completely eliminates all midline glia and dramatic loss of VUM neurons. Results indicate all midline glia are capable of undergoing cell death and suggests the midline neurons and glia may have different sensitivities to rpr and W expression. Simultaneous expression of BacA\p35Scer\UAS.cHa causes no ectopic cell death demonstrating that midline cell death induced by rpr and W require the functions or one or more caspases.
In WScer\UAS.cZa; BacA\p35Scer\UAS.cHa; Scer\GAL4en-e16E animals, increased BrdU incorporation is seen in the posterior compartment of the wing disc and in anterior compartment cells adjacent to the compartment boundary.