The expression of Tip60E431Q.UAS under the control of Scer\GAL4NP1624 leads to testes showing increased numbers of cyst stem cells (Zfh-1-positive cells), associated with cyst stem cell differentiation defects, as illustrated by a high proportion of cells abnormally co-expressing Zfh-1 (marker of cyst stem cells/early cyst cells) and Eya (marker of later stage cyst cells). About half of testes also show germ cell expansion, with some testes also showing structures that resemble stem cell, GB and spermatogonial tumors. Additionally, some testes show cells expressing both germ cell markers (Vasa) and cyst cell markers (Zfh-1 or Yan).
Flies expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4ey-OK107 show a significant reduction in total mushroom body size as compared to wild type. These flies also show a loss of mushroom body neuroadaptive response to environmental enrichment in the first 5 days of adulthood, in contrast to wild type flies, which exhibit an increase in mushroom body size when housed with other flies during the first 5 days of adulthood, versus those kept in isolation.
Scer\GAL4ey-OK107>Tip60E431Q.Scer\UAS male flies show normal learning in a conditioned courtship suppression assay (placed with a mated female for 60 minutes during the training phase), with a significant drop in the courtship index when comparing the first 10 minutes to the final 10 minutes (similar to wild type). Two minutes after training, flies are placed with a virgin female and courtship is assessed for 10 minutes: trained controls show significantly reduced courtship index when compared to sham (untrained males), whereas Scer\GAL4ey-OK107>Tip60E431Q.Scer\UAS males show no significant drop (immediate-recall memory defect).
Stereotyped morphology of the mushroom bodies looks normal in Scer\GAL4ey-OK107>Tip60E431Q.Scer\UAS third instar larvae, but severe axonal defects are detected in adult flies: alpha/alpha' lobes are thinner and there is a significant decrease in the area of all lobes (alpha/beta, alpha'/beta', gamma) compared to controls.
Third instar larvae expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4elav-C155 are significantly less mobile and perform significantly fewer full body peristaltic contractions in a locomotor assay compared to controls. The time taken for the larvae to right themselves after being turned ventral side up is also significantly longer than normal.
Larvae expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4elav-C155 show abnormal aggregations of vesicles on motor neurons extending from the central nervous system.
Expression of Tip60E431Q.Scer\UAS under the control of Scer\GAL4P2.4.Pdf has no adverse effect on the axonal pattern development in third instar larvae but leads to loss of s-LNv terminal synaptic arborization and reduced axon length in the adult brain compared to controls. It also causes decrease in nighttime sleep and its disruption and fragmentation as well as daytime sleepiness but the flies retain normal circadian activity pattern.
Moderate levels of apoptotic cell death are observed in third instar larval brains expressing Scer\GAL4179Y>Tip60E431Q.Scer\UAS.
Expression of Tip60E431Q.Scer\UAS at 25[o]C from late embryonic development to adulthood, under the control of Scer\GAL4337Y is lethal. The developmental stage when lethality occurs varies between individual fly lines, with the majority of lethality occurring during the late pupal stage for line A, and during the late second instar larvae stages for lines B and C. Similar results are seen upon expression with Scer\GAL4Act5C.PI. Endogenous levels of acetylated histone His4 are significantly depleted in in Scer\GAL4337Y-->Tip60E431Q.Scer\UAS flies.
Expression of Tip60E431Q.Scer\UAS in the nervous system, under the control of Scer\GAL4elav.PLu (pan-neuronally), Scer\GAL4179Y (pan-neuronally) or Scer\GAL460IIA (specifically in the brain and CNS), causes a reduction in viability. This viability varies between 0-25% for expression under the control of Scer\GAL4elav.PLu (depending on the transgenic line), 30% for Scer\GAL4179Y, and 40% for Scer\GAL460IIA, respectively.
Pre-synaptic expression of Tip60E431Q.Scer\UAS under the control of Scer\GAL4elav-C155 results in pupal lethality. There is a significant increase in the total number of synaptic boutons in Tip60E431Q.Scer\UAS-expressing larvae, when compared to wild-type controls. There is a substantially larger expansion of type-I boutons compared to type-Ib in these mutants. There is also a slight increase in the number of satellite boutons in these mutants compared to controls. This increase in bouton number is accompanied by an excess of futsch-associated microtubule looping and splaying. Neuromuscular junctions in these larvae exhibit a reduction in acetylated microtubule staining, compared to wild-type.
Post-synaptic expression of Tip60E431Q.Scer\UAS under the control of Scer\GAL4Mef2.247 results in pupal lethality. Mutant larvae exhibit a significant reduction in bouton number, specifically in 1s boutons, as well as the absence of satellite boutons.
Scer\GAL4NP1624, Tip60E431Q.UAS has neoplasia | spermatogenesis phenotype, enhanceable by E(Pc)[+]/E(Pc)w3
Scer\GAL4elav-C155, Tip60E431Q.UAS has abnormal locomotor behavior | third instar larval stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4elav-C155
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has abnormal neuroanatomy | adult stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Scer\GAL4179Y, Tip60E431Q.UAS has increased mortality during development phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4179Y
Scer\GAL4179Y, Tip60E431Q.UAS has increased cell death | third instar larval stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4179Y
Scer\GAL4337Y, Tip60E431Q.UAS has increased mortality during development phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4337Y
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has abnormal neuroanatomy | adult stage phenotype, non-enhanceable by Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Scer\GAL4elav-C155, Tip60E431Q.UAS has abnormal locomotor behavior | third instar larval stage phenotype, suppressible | partially by Scer\GAL4elav-C155/Tip60UAS.cLa
Scer\GAL4337Y, Tip60E431Q.UAS has lethal phenotype, non-suppressible by Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4337Y
Scer\GAL4337Y, Tip60E431Q.UAS has majority die during third instar larval stage phenotype, non-suppressible by Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4337Y
Scer\GAL4337Y, Tip60E431Q.UAS has lethal phenotype, non-suppressible by Hsap\APP695ΔCT.UAS.Tag:MYC/Scer\GAL4179Y
Scer\GAL4337Y, Tip60E431Q.UAS has majority die during third instar larval stage phenotype, non-suppressible by Hsap\APP695ΔCT.UAS.Tag:MYC/Scer\GAL4179Y
Tip60E431Q.UAS, Scer\GAL4ey-OK107 is an enhancer of abnormal neuroanatomy | adult stage phenotype of Hsap\APP695.UAS.Tag:MYC, Scer\GAL4ey-OK107
Tip60E431Q.UAS, Scer\GAL4elav-C155 is an enhancer of abnormal locomotor behavior | third instar larval stage phenotype of Hsap\APP695.UAS.Tag:MYC, Scer\GAL4elav-C155
Tip60E431Q.UAS, Scer\GAL4ey-OK107 is a non-suppressor of abnormal learning | adult stage phenotype of Hsap\APP695.UAS.Tag:MYC, Scer\GAL4ey-OK107
Tip60E431Q.UAS, Scer\GAL4ey-OK107 is a non-suppressor of abnormal memory | adult stage phenotype of Hsap\APP695.UAS.Tag:MYC, Scer\GAL4ey-OK107
Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4ey-OK107, Tip60E431Q.UAS has abnormal memory | adult stage phenotype
Hsap\APP695.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has abnormal sleep | adult stage phenotype
Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has abnormal sleep | adult stage phenotype
Scer\GAL4NP1624, Tip60E431Q.UAS has male germline stem cell | increased number phenotype, enhanceable by E(Pc)[+]/E(Pc)w3
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has s-LNv Pdf neuron | adult stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has axon | adult stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Scer\GAL4179Y, Tip60E431Q.UAS has embryonic/larval brain | third instar larval stage phenotype, enhanceable by Hsap\APP695.UAS.Tag:MYC, Scer\GAL4179Y
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has s-LNv Pdf neuron | adult stage phenotype, non-enhanceable by Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Scer\GAL4P2.4.Pdf, Tip60E431Q.UAS has axon | adult stage phenotype, non-enhanceable by Hsap\APP695ΔCT.UAS.Tag:MYC, Scer\GAL4P2.4.Pdf
Tip60E431Q.UAS, Scer\GAL4ey-OK107 is an enhancer of adult mushroom body phenotype of Hsap\APP695.UAS.Tag:MYC, Scer\GAL4ey-OK107
Co-expression of Tip60E431Q.Scer\UAS exacerbates axonal defects in all three mushroom body lobes, resulting in a lack of structural consistency of the mushroom body in flies expressing Hsap\APP695.Scer\UAS.T:Hsap\MYC under the control of Scer\GAL4ey-OK107.
Co-expression of Tip60Scer\UAS.cLa (but not Tip60E431Q.Scer\UAS) suppresses learning and memory defects in Scer\GAL4ey-OK107>Hsap\APP695.Scer\UAS.T:Hsap\MYC males. Co-expression of Tip60Scer\UAS.cLa or Tip60E431Q.Scer\UAS does not affect learning but abolishes short-term memory of courtship suppression in Scer\GAL4ey-OK107>Hsap\APP695ΔCT.Scer\UAS.T:Hsap\MYC males.
Larvae co-expressing both Tip60E431Q.Scer\UAS and Hsap\APP695.Scer\UAS.T:Hsap\MYC under the control of Scer\GAL4elav-C155 show a synergistic enhancement of the locomotor defects that are seen in larvae expressing either Tip60E431Q.Scer\UAS or Hsap\APP695.Scer\UAS.T:Hsap\MYC alone. In addition, the formation of axonal vesicle accumulations is exacerbated in the double mutants.
The defects in the axonal pattern of s-LNv neurons in brains characteristic for adult flies expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4P2.4.Pdf are exacerbated further by co-expression of Hsap\APP695.Scer\UAS.T:Hsap\MYC but not by co-expression of Hsap\APP695ΔCT.Scer\UAS.T:Hsap\MYC. Both flies co-expressing Tip60E431Q.Scer\UAS with Hsap\APP695.Scer\UAS.T:Hsap\MYC or with Hsap\APP695ΔCT.Scer\UAS.T:Hsap\MYC show reduction in nighttime sleep with concomitant increase in daytime sleep (also seen in flies expressing either Tip60E431Q.Scer\UAS or Hsap\APP695.Scer\UAS.T:Hsap\MYC alone).
Co-expression of Tip60E431Q.Scer\UAS and Hsap\APP695.Scer\UAS.T:Hsap\MYC both under the control of Scer\GAL4337Y results in 0% viability, with lethality occurring during the early second instar larval stage. Additionally, hatching of 100% of these larvae is delayed by 24-48 hours.
Tip60E431Q.UAS is rescued by Scer\GAL4elav-C155/Tip60UAS.cLa
Tip60E431Q.UAS is rescued by Tip60UAS.cLa/Scer\GAL4P2.4.Pdf
Tip60E431Q.UAS is rescued by Scer\GAL4337Y/Tip60UAS.cLa
Co-expression of Tip60Scer\UAS.cLa rescues the locomotor behaviour defects seen in third instar larvae expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4elav-C155.
The loss of s-LNv terminal synaptic arborization and reduced axon length in the adult brain as well as the sleep defects characteristic for flies expressing Tip60E431Q.Scer\UAS under the control of Scer\GAL4P2.4.Pdf is rescued by co-expression of Tip60Scer\UAS.cLa.
Co-expression of Tip60Scer\UAS.cLa rescues the lethality associated with the expression of Tip60E431Q.Scer\UAS, when both transgenes are under the control of Scer\GAL4337Y.
