put10460 third instar larval C4da neurons do not show any obvious defects in the dendritic arborization, as compared to controls.
put10460 somatic clones in the pupal retina show mild defects in the arrangement of inter-ommatidial pigment cells.
put10460/+ flies display a wild-type bristle pattern.
Embryos laid from put10460 germ-line clones and fertilised with wild-type sperm are weakly ventralised.
put135/put10460 males raised to adulthood at 18oC and then shifted to 22oC for 7 days have normal numbers of germline stems cells per testis. However, one week at 29oC is sufficient to almost completely eliminate germline stem cells in these animals. No increase in apoptosis occurs of germline stem cells occurs in these testes during culture at 29oC, suggesting that loss is due to differentiation of stem cells. Clones of male put10460 homozygous germline stem cells are still present in 15% of testes one week after clone induction and none are present two weeks after clone induction. This is in contrast to wild-type control clones, which are present in 82% of testes one week after clone induction and 64% two weeks after clone induction.
The average number of crystal cells per embryo is reduced (to 13.5) in homozygous stage 13-14 embryos compared to wild type (36 +/- 2.2 cells on average). The number of plasmatocytes is also significantly reduced compared to wild-type embryos.
At eclosion put10460/put135 adult females raised at 18oC have normal numbers of germ-line stem cells, but after 2 days at 29oC 53% of germaria in these flies have one or no germline stem cells.
put135/put10460 males shifted to the restrictive temperature of 29oC after eclosion have smaller and thinner testes than control males after 7 days at 29oC. The testes also show an apparent reduction in the number of early germ cells, particularly spermatogonia and germ line stem cells.
The tracheal systems of put10460 mutant embryos fail to form dorsal branches.
Clones of put135/put10460 induced in the pleura cause no mutant phenotype.
put135/put10460 animals shifted from 16 to 28oC 6 days after egg deposition show loss of the dorsocentral macrochaetae, although lateral macrochaetae and the field of microchaetae remain. There is also loss of wing tissue. put135/put10460 animals shifted from 16 to 28oC during larval 7 days after egg deposition produce extra dorsocentral macrochaetae.
Homozygous clones at the posterior margin of the eye disc do not show photoreceptor differentiation.
At 25oC, 18% of homozygous embryos have a strong dorsal-open phenotype (severe head defects and a large hole in the dorsal cuticle), 43% have an intermediate phenotype (the dorsal cuticle is noticeably reduced, and there is increased curvature in the abdominal segments, resulting in a "tail-up" phenotype) and 39% have a weak phenotype. put10/put10460 adults are fully viable at 25oC, but show defects in wing venation in 28% of cases.
put10460/put135 animals raised at the permissive temperature (18oC) and then shifted to the nonpermissive temperature (29oC) at about 48 hours before pupariation lack the dorsocentral sensory mother cells in the thoracic discs. put10460/put135 animals raised at the permissive temperature (18oC) and then shifted to 25oC have one or a few extra sensory mother cells in the thoracic discs.
Clonal analysis in the germarium reveals that mutant stem cells are lost more rapidly than wild type, though there is no effect on the formation of 16 cell cysts or their subsequent development. Stem cell half life is 0.9 weeks (wild type being 4.6 weeks). Stem cell division rate relative to control is 0.36. Cysts contain the normal 16 cells, including and oocyte.
Embryos exhibit a partial dorsal closure phenotype.
Large mutant clones covering almost the entire adult eye are generally wild type (with a small amount of wild type tissue at the posterior border); ommatidia are normal, occasional vacuoles between the ommatidia are observed. The head of an adult fly with mutant clones has no eye structures, only head cuticle is present where the eye normally forms. Clones in the imaginal disc anterior to the morphogenetic furrow have no detectable phenotype. Clones posterior to the furrow are also normal demonstrating the furrow can progress normally through and beyond a mutant clone. Within clones traversing the furrow neuronal differentiation is retarded, slowing of the furrow is more marked in the middle of the clone suggesting partial rescue by surrounding wild type tissue. Clones at the posterior margin of the eye discs can cause local overproliferation and block neuronal differentiation. Neuronal differentiation does occur in lateral clones.
Wing clones induced in mid-to-late third instar larvae are small due to their late induction time but cause visible mutant phenotypes: gaps, splits, vein indentations and additional vein material abutting normal veins.
Homozygous embryos exhibit a dorsal open phenotype. Germline clones produce eggs with patterning defects: embryos are ventralised.
put135/put10460 transheterozygous pharate adults heat shocked during late first and early second instar have legs in which dorsal structures are absent and ventral structures are duplicated. On the distal tibia of the second leg there are two apical bristles which are indicators of the ventral-most cell fate in leg discs. Pharate adults exhibit reduced or missing eyes and duplicated antenna.
Dorsal open phenotype and head defects become more severe at the higher temperatures. Heterozygotes with put51 and put88 are lethal at high temperatures, at low temperatures individuals survive and exhibit ectopic wing venation in the region of the posterior cross vein. Escaper females produce ventralised embryos.