put135 heterozygous females show the expected mating-induced increase in germline stem cells in the germarium.
The dorsal branches of the dorsal trunk of the tracheal system are absent in put135 embryos.
Tracheal dorsal branches do not form in put135 embryos.
Six days after induction of intestinal stem cell (ISC) clones, the number of cells in put135 mutant clones is significantly higher than in wild-type clones. After 24-h bleomycin treatment, the mean cell number per put135 mutant clone is larger than the mean cell number per wild-type clone.
At permissive temperatures of 18-20oC, put135/put62 embryos do not show significant axon guidance defects. At 23-25oC, these embryos show some guidance defects, affecting the intersegmental nerve and the SNa and SNb. At temperatures above 25oC, embryos of this genotype exhibit gross head involution and dorsal closure defects.
When flies with dorsal cluster neuron put135 clones are reared at the restrictive temperature, fewer neurites from these clones reach the contralateral chiasm/medulla than when these flies are reared at the permissive temperature. Mosaic analysis shows that flies must be cultured at the restrictive temperature as 3rd-instar larvae to show this effect.
When neutral marked clones are induced in the ovary, the proportion of germaria carrying marked somatic stem cells 3 weeks after clone induction is around 70% of that seen on week after clone induction. However, for put135 homozygous clones, the equivalent figure is around 20%.
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. Clones of male put135 homozygous germline stem cells are still present in 21% 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.
put135/put51 animals raised at 18oC have reduced body and muscle size, but the number of synaptic boutons in these mutants is normal.
The average number of crystal cells per embryo is reduced in homozygous stage 13-14 embryos (raised at the nonpermissive temperature of 30oC) compared to wild type.
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.
Homozygous germ line clones (either germ line stem cells or spermatocytes) are generated in the testis, but they do not persist to the same extent as wild-type control clones. 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.
Mutant clones fail to exhibit any visible defects in mushroom body remodelling.
The Malpighian tubules of homozygous embryos vary in width along their length. The tubules grow to approximately normal size but do not complete cell rearrangement. When the tubules are fully developed, the distal segments are elongated, but the proximal 80% or more of the tubules are unelongated. The width of the tubules in some portions appears greater than the width of normal tubules before elongation, but the lumens are not enlarged.
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 embryos show bunching of epidermal segments at the end of dorsal closure.
Lethal in transheterozygous combination with putD13 or putD18.
put10/put135 adults are viable, and sometimes show defects in wing venation. Temperature shift experiments using put135/put88 animals indicate that there is an absolute requirement for zygotic put function between 8 and 12 hours of embryogenesis. put62/put135 flies are inviable at 25oC and fully viable at 18oC. put62/put135 flies maintained at 18oC throughout embryogenesis and then shifted to 25oC show a marked reduction in viability and are grossly deformed. These animals all have notal defects, usually medial notal clefts, and have leg defects in 99% of cases, which include truncations, bifurcations and abnormal twists. Distal pattern elements are usually deleted in at least one limb. Duplications of sex combs are often seen on the forelegs. 97% of these flies have gross eye and antennal defects. The eyes are highly disorganised and have a reduced number of ommatidia and bristles. Antennal defects include duplications and deletions of distal pattern elements.
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 0.41 weeks (wild type being 4.6 weeks). Stem cell division rate relative to control is 0.37.
Somatic clones in the male testis produce overproliferating germ cells, a cyst contains many more than 16 cells that are much smaller than wild type. Germ line clones do not show overproliferation of the germ cells.
Embryos exhibit a partial dorsal closure phenotype.
All dorsal branches of the tracheal system are absent and defects in the ventral branches are occasionally observed when embryos are collected at 29oC.
Mutant embryos show absence of the dorsal branch and reduced lateral trunks. The visceral branch forms normally.
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.
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.
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. sax1/sax2 put135/put+ individuals lack the posterior cross vein.
Severe head defects and dorsal open phenotype. Embryos lacking maternal and paternal put contribution are completely ventralised.
put135 embryos are open anterodorsally.
embryonic lethal Dorsal part of embryo open.