fertile (with Df(2R)Exel7095), with LpinUAS.cUa, Scer\GAL4αTub84B.PL
lethal | P-stage (with Df(2R)Exel7095)
Lpine00680/Df(2R)Exel7095 larvae exhibit an increase in larval lethality, underdeveloped fat bodies, defective fat body cell morphology with increased cell and nucleus size, and decreased fat droplet size, as compared to controls.
Animals homozygous for Lpine00680 are viable, but many animal die during late larval and pupal development, and only a few adult flies develop. The mutant adult flies show strongly reduced fertility, laying less than 10% of the eggs produced by their heterozygous siblings. Lpine00680 mutant larvae show delayed development, entering entering the wandering stage about 2 days later than heterozygous control animals. Homozygous Lpine00680 results in impaired eclosion. 59% of homozygous Lpine00680 animals die in the pupal case, many after reaching the pharate adult stage. Dead homozygous Lpine00680 pharate adults do not show obvious morphological defects. The Lpine00680 mutant and wild-type control pupae are similar in size. Some adults die while trying to eclose, suggesting that these animals lack the energy to emerge fully from the pupal case.
Many of the Lpine00680/Df(2R)Exel7095 mutant larvae look transparent, indicating a dramatic reduction in the amount of fat tissue. The function of the fat body is found impaired in Lpine00680/Df(2R)Exel7095 mutants. Dissection of the larval fat body of Lpine00680/Df(2R)Exel7095 animals reveals that while the total mass of fat tissue is greatly reduced, individual fat body cells are highly enlarged. The nuclei of these cells are enlarged as well or have apparently undergone fragmentation. On average, the sizes of both cells and cell nuclei are increased about two-fold. Some mutant cells reach a diameter of 160 μm. In contrast, the cells and nuclei of the larval salivary glands do not show a significant change in size. The shape of the Lpine00680/Df(2R)Exel7095 mutant fat body cells is different from that of wild-type controls. The mutant cells are more rounded than control cells and, in some regions of the fat body, appear detached from their neighbours, indicating impaired cell adhesion. The large fat droplets which are plentiful in wild-type fat body cells are scarce in the Lpine00680/Df(2R)Exel7095 mutant fat bodies. This reduction in lipid droplet size is accompanied by a significant overall reduction in triglyceride levels.
The level of the ketone β-hydroxybutyrate is almost two-fold higher in Lpine00680/Df(2R)Exel7095 mutants than in wild-type animals.
The average length of Lpine00680/Df(2R)Exel7095 mutant pupae are similar to that of wild-type.
Only 23% of the Lpine00680/Df(2R)Exel7095 larvae reach the wandering third instar larval stage.
79% of Lpine00680/Df(2R)NCX10 and more than 99% of the Lpine00680/Df(2R)Exel7095 animals die during the pupal stage.
Lpine00680/Df(2R)Exel7095 mutant fat body cells contain autophagosomes and autolysosomes, but the matrix of these vesicles is considerably less dense than the matrix in wild-type control cells. Furthermore, the cells contain misshapen nuclei with nuclear envelopes that form projections or involutions or appear ruptured. The mutant cells contain unusually structured, and apparently defective, mitochondria. In contrast to the elongated crista-rich mitochondria of control cells, these mitochondria are rounded and contain few cristae. In some cases, both the outer and the inner mitochondrial membranes are ruptured.
Df(2R)Exel7095/Lpine00680 has lethal phenotype, non-suppressible by wun2UAS.Tag:MYC/Scer\GAL4αTub84B.PL
Df(2R)Exel7095/Lpine00680 has lethal phenotype, non-suppressible by wunUAS.cZa/Scer\GAL4αTub84B.PL
Constitutive expression of BacA\p35Scer\UAS.cHa under the control of Scer\GAL4r4 in the fat bodies of Lpine00680/Df(2R)Exel7095 mutants further enhances the lethality observed for Lpine00680/LpinGD14004 animals. The few surviving wandering larvae are characterised by severe lipodystrophy and dramatically oversized and rounded fat body cells. Some of these cells show fragmented nuclei and are indistinguishable from Lpine00680/Df(2R)Exel7095 mutant cells in the absence of BacA\p35Scer\UAS.cHa.
Df(2R)Exel7095/Lpine00680 is rescued by Scer\GAL4αTub84B.PL/LpinUAS.cUa
Df(2R)Exel7095/Lpine00680 is rescued by LpinUAS.cSa/Scer\GAL4r4
Df(2R)Exel7095/Lpine00680 is rescued by Scer\GAL4r4/LpinΔNLS.UAS
Df(2R)Exel7095/Lpine00680 is not rescued by LpinΔPAP.UAS/Scer\GAL4r4
Expression of LpinScer\UAS.cSa or LpinΔNLS.Scer\UAS, but not LpinΔPAP.Scer\UAS, under the control of Scer\GAL4r4 rescues the defects in fat body cell morphology and lipid droplet formation seen in Lpine00680/Df(2R)Exel7095 larvae. Expression of LpinΔPAP.Scer\UAS under the control of Scer\GAL4r4 increases larval lethality seen in these flies.