UASt regulatory sequences drive expression of an inverted repeat.
viable, with Scer\GAL471B
wing, with Scer\GAL471B
Expressing TaoGD8262 under the control of Scer\GAL4da.PU results in a smaller ventral nerve cord with less-developed axons in segmental nerves, changed NMJ morphology and a significantly decreased number of NMJ boutons per muscle section in third instar larvae compared to wild-type controls.
Expressing TaoGD8262 under the control of Scer\GAL4Toll-6-D42 results in disturbed mitochondrial distribution and significantly decreased average mitochondrial size in axons of motor neurons in third instar larvae compared to wild-type controls.
Expressing TaoGD8262 under the control of Scer\GAL4insc-Mz1407 (in combination with a Dicer-2 transgene to enhance RNAi efficiency) results in premature type I neuroblast reactivation and a significant increase in neuroblast diameter in late-first/early-second instar larval ventral nerve cords and brains, concomitant with an overall volume increase in the central nervous system. Neuroblast proliferation and central nervous system volume are significantly increased compared to controls throughout larval life, from mid-second instar, to late-second/early-third and third instar stages. At 24 hours after puparium formation, a significantly higher number of persistent type I neuroblasts in the ventral nerve cord, together with an increased neuroblast diameter, indicate that neuroblast cell-cycle exit and differentiation are delayed compared to controls.
Type I neuroblasts in somatic clones expressing TaoGD8262 under the control of Scer\GAL4Tub.PU (in combination with a Dicer-2 transgene to enhance RNAi efficiency) display significant increases in cell diameter, proliferation (indicated by EdU incorporation), cell-cycle speed and frequency of cell division in the ventral nerve cord of third instar larvae. As a result, TaoGD8262-expressing clones have an overall greater number of neurons, but a reduced neuronal cell size. The absolute numbers of both early- and late-born neurons are significantly increased, together with an expanded proportion of early-born neurons compared to controls, without disrupting temporal identity. Similar to wild-type controls, each somatic clone has only one neuroblast, the neuroblast undergoes asymmetric divisions, ganglion mother cells divide symmetrically, there is no neuronal proliferation, and cell death levels are unaffected. In the adult ventral nerve cord, somatic clones lack persistent neuroblasts and show no sign of proliferation.
Expressing TaoGD8262 under the control of Scer\GAL4NP1316 (in combination with a Dicer-2 transgene to enhance RNAi efficiency) results in a significant volume increase of the central nervous system at 24 hours after puparium formation.
Somatic clones expressing TaoGD8262 under the control of Scer\GAL4NP1316 (in combination with a Dicer-2 transgene to enhance RNAi efficiency) show a significantly increased mitotic index of type I neuroblasts in the ventral nerve cord of third instar larvae compared to controls. Neuroblasts maintain cell polarity typical of wild-type controls.
Expressing TaoGD8262 under the control of Scer\GAL4wor.PA in combination with Scer\GAL80ase.PN (to restrict expression to type II neuroblasts) and a Dicer-2 transgene (to enhance RNAi efficiency) results in a significant decrease in the volume of type II lineages in brain lobes compared to controls. A similar volume decrease is observed in somatic clones expressing TaoGD8262 under the control of Scer\GAL4Tub.PU. Conversely, somatic clones of type I lineages in the ventral nerve cord (but not in the central brain) show a significant increase in volume compared to controls.
Adult flies expressing TaoGD8262 in the intestinal stem cells (ISCs) and enteroblasts (EBs) of the adult midgut for 7 days under the control of Scer\GAL4esg-NP5130 and Scer\GAL80ts.αTub84B contain an increased number of esg-positive cells in the midgut. These cells are often clustered, with aberrant morphology. Their cell size is increased, with many similar in size to enterocytes (ECs). Mitotic activity (PH3 staining) is significantly increased compared to controls. There is an increased in the number of Dl-positive ISC-like cells, EBs and enteroendocrine cells (ees).
Intestinal stem cell clones expressing TaoGD8262 under the control of Scer\GAL4Act.PU are significantly larger than controls.
Expression of TaoGD8262 under the control of Scer\GAL4ap-md544 causes massive overgrowth of third instar wing imaginal discs compared to wild type.
In adult wings from animals expressing TaoGD8262 under the control of Scer\GAL4dpp.blk1, extra folds and vein material are present. The area between veins three and fours shows a statistically significant increase in size compared to wild type wings.
Expression of TaoGD8262 in the dorsal compartment of the eye using Scer\GAL4mirr-DE results in an increase in the size of the eye imaginal discs. The dorsal compartment is grossly overgrown relative to both the ventral compartment and to wild type controls.
An increased number of interommatidial cells is seen in pupal eyes when TaoGD8262 is expressed under the control of Scer\GAL4GMR.PF. Unlike in wild type, the third instar larval eye disc cells continue to divide posterior to the second mitotic wave.
Expression of TaoGD8262 under the control of Scer\GAL4en-e16E results in lethality before adult stages. The wing discs are increased in size compared to controls.
Expression of TaoGD8262 under the control of Scer\GAL4elav-C155 results in ethanol-induced hyperactivity that is reduced compared to controls.
Expression of TaoGD8262 in the mushroom bodies under the control of Scer\GAL4ey-OK107 often leads to loss of the α/β mushroom body lobes. When theses lobes are absent, a FasII-positive ball-like structure is present adjacent to the mushroom body calices. Increasing the strength of TaoGD8262 expression (using Dcr-2Scer\UAS.cDa) reveals defects in mushroom body morphology in all brains examined. All α/β lobes examined are disrupted to some degree, most being shorter or thinner than normal. Some α' lobes are fully formed, but no normal β' lobes are seen. The γ lobes present are present in most brains but are often diffuse and shortened.
Expression of TaoGD8262 (with Dcr-2Scer\UAS.cDa) under the control of Scer\GAL4ey-OK107 the third instar larvae mushroom bodies show no obvious defects in morphology. However, by 24h after pupariation defects are apparent in the dorsal lobes, which were greatly reduced or absent.
Expression of TaoGD8262 (with Dcr-2Scer\UAS.cDa) in the mushroom bodies using Scer\GAL4ey-OK107 results in a reduction in ethanol-induced hyperactivity. The ellipsoid body is intact in these flies.
Expression of TaoKK108458 under the control of the wing driver Scer\GAL471B at 25[o]C results in viable adults, however the overgrown and crumpled wing phenotype precludes accurate measurement of adult wing size. At 18[o]C a 7% increase in wing size is seen compared to controls.
Adults expressing TaoGD8262 under the control of Scer\GAL4elav.PLu (in the presence of Dcr-2Scer\UAS.cDa to increase the efficiency of RNAi) show significantly reduced avoidance of noxious temperature (46[o]C) compared to control flies.
Adults expressing TaoGD8262 under the control of Scer\GAL4elav.PLu (in the presence of Dcr-2Scer\UAS.cDa to increase the efficiency of RNAi) show no significant change in the kinetics of high temperature (46[o]C) induced paralysis compared to control flies.
Expression under the control of Scer\GAL4Mef2.PR results in late pupal lethality.
Scer\GAL4insc-Mz1407, TaoGD8262 has abnormal neuroanatomy | first instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has abnormal neuroanatomy | second instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has abnormal size | first instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has abnormal size | second instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4en-e16E, TaoGD8262 has lethal phenotype, suppressible | partially by Scer\GAL4en-e16E/wtsUAS.L.Tag:MYC
Scer\GAL4en-e16E, TaoGD8262 has abnormal size | third instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4en-e16E
TaoGD8262/Scer\GAL4da.PU, Scer\GAL80ts.αTub84B is a non-enhancer of lethal - all die during pupal stage | heat sensitive phenotype of Scer\GAL4da.PU, Scer\GAL80ts.αTub84B, fzrRNAi.UAS.WIZ
Scer\GAL4en-e16E/TaoGD8262 is a suppressor | partially of visible | adult stage phenotype of Scer\GAL4en-e16E, wtsUAS.L.Tag:MYC
Scer\GAL4en-e16E/TaoGD8262 is a suppressor | partially of abnormal size | adult stage phenotype of Scer\GAL4en-e16E, wtsUAS.L.Tag:MYC
TaoGD8262/Scer\GAL4da.PU, Scer\GAL80ts.αTub84B is a non-suppressor of lethal - all die during pupal stage | heat sensitive phenotype of Scer\GAL4da.PU, Scer\GAL80ts.αTub84B, fzrRNAi.UAS.WIZ
Scer\GAL4insc-Mz1407, TaoGD8262 has embryonic/larval central nervous system | first instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has embryonic/larval central nervous system | second instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has type I neuroblast | first instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4insc-Mz1407, TaoGD8262 has type I neuroblast | second instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4insc-Mz1407
Scer\GAL4Act.PU, TaoGD8262 has intestinal stem cell | somatic clone phenotype, suppressible by ykiB5
Scer\GAL4NP5130, Scer\GAL80ts.αTub84B, TaoGD8262 has adult midgut phenotype, suppressible by ykiHMS00041/Scer\GAL4NP5130, Scer\GAL80ts.αTub84B
Scer\GAL4en-e16E, TaoGD8262 has wing disc | third instar larval stage phenotype, suppressible by ykiKK109756/Scer\GAL4en-e16E
Scer\GAL4ey-OK107, TaoGD8262 has adult mushroom body alpha-lobe phenotype, suppressible | partially by par-1W3/par-1[+]
Scer\GAL4ey-OK107, TaoGD8262 has adult mushroom body beta-lobe phenotype, suppressible | partially by par-1W3/par-1[+]
Scer\GAL4en-e16E/TaoGD8262 is a suppressor | partially of wing phenotype of Scer\GAL4en-e16E, wtsUAS.L.Tag:MYC
Expression of ykiHMS00041 suppresses the increase in esg-positive cells and mitosis seen when TaoGD8262 is expressed in adult midguts under the control of Scer\GAL4esg-NP5130 and Scer\GAL80ts.αTub84B.
ykiB5 suppresses the increase in intestinal stem cell clone size seen when TaoGD8262 is expressed under the control of Scer\GAL4Act.PU.
Expression of wtsScer\UAS.L.T:Hsap\MYC partially suppresses the lethality seen when TaoGD8262 is expressed under the control of Scer\GAL4en-e16E.
Expression of ykiKK109756 suppresses the wing disc over-proliferation phenotype seen when TaoGD8262 is expressed under the control of Scer\GAL4en-e16E at 18[o]C.
Expression of TaoGD8262 partially suppresses the wing under-proliferation seen when wtsScer\UAS.L.T:Hsap\MYC is expressed under the control of Scer\GAL4en-e16E.
One copy of par-1W3 partially suppresses the adult mushroom body morphology defects seen when TaoGD8262 is weakly expressed in mushroom bodies under the control of Scer\GAL4ey-OK107.
