The expression of
BacA\p35UAS.cHa from the second instar larval stage under the combined control of
Scer\GAL4ap-md544 and Gal80[ts] does not significantly affect the size of the third instar larval wing disc dorsal compartment, nor their average centrosome number per cell, as compared to controls.
The expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4pnt-14-94 does not lead a significant change in the number of type II neuroblasts per third instar larval brain lobe, as compared to controls.
The expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4ey.PU does not affect the mitotic index in the third instar larval eye disc, as compared to controls.
Upon exposure to ionizing radiation, individuals expressing
BacA\p35UAS.cHa under the control of
Scer\GAL4en.PU display wing disc posterior compartments that are significantly enlarged, present basal cell delamination and present
BacA\p35UAS.cHa-expressing cell far away from the domain of expression; these phenotypes are not observed under non-irradiated conditions.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Sgs3.PD does not block the formation of the large acidic structures which normally occurs in salivary gland cells approximately 1.5 hours after head eversion, but caspase activation at 2 hours after head eversion is blocked. This does not prevent breakdown of the salivary gland tissue, as no persistent salivary gland phenotype is seen at 24 hours after puparium formation.
Expression of five copies of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4VGlut-OK371 does not alter the rate of axon degeneration in the L1 wing vein neurons following axotomy compared to wild type.
Egg chambers expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4os.cXa have an increased number of polar cells per cluster compared to wild type; all wild type egg chambers have two polar cells per cluster, whereas 35% of polar cell clusters contain more than two cells.
11.1 +/- 1.3 surviving EW3-sib cells are seen in larvae expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4en.PU (these cells die during embryogenesis in wild type).
Expression of
BacA\p35UAS.cHa under the control of
Scer\GAL4NP5130 has no obvious effect on the proportion of Esg-positive progenitors in the adult posterior midgut.
Expression of
BacA\p35Scer\UAS.cHa in larval wing disc cells, under the control of
Scer\GAL4ptc-559.1, do not display invasive cell behaviour, but contain occasional cells with processes extended towards the posterior compartment.
Blocking caspase activity in neurons through expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4elav.PU results in embryos with a reduced volume of apoptotic particles and reduced engulfment of apoptotic neurons compared to controls.
Males expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4en.PU display mis-oriented adult genitalia. During pupal development, the genital disc does not undergo full rotation. Rotation commences as normal and the initial average velocity is comparable to control flies. However rotation stops before it is complete and the acceleration-stage of rotation is lower compared to controls. The inner ring of the disc rotates normally in these animals, but the rotation of the outer ring is impaired.
Adult brains of animals containing clones expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4tub.PU do not show cell accumulation in the medulla cortex or alteration in the medulla neuropil lamination.
Class III neurons overexpressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL419-12 do not undergo apoptosis and are still present by 18 hours after puparium formation.
Blocking apoptosis through expression of
BacA\p35Scer\UAS.cHa in the wing margin cells under the control of
Scer\GAL4wg-IS650 (with
Scer\GAL80ts.αTub84B blocking expression at other times), results in survival of the flat cells and no ziz-zag alignment of the wing margin cells, as found in wild-type.
Expression of
BacA\p35Scer\UAS.cHa in the pupal wing, under the control of
Scer\GAL4wg-IS650 (with
Scer\GAL80ts.αTub84B blocking expression at other times), results in misalignment of hair shafts, with some aligned in an almost side-by-side manner by chance. Consistent with this, in the adult wing expressing
BacA\p35Scer\UAS.cHa, the alternate projection of the hairs is disturbed, and hairs from the dorsal and ventral compartments overlap. Blocking apoptosis also affects the position of the sensory bristles in the anterior wing margin, leading to overlap of the double-row hairs, although the phenotypic relationship between the anterior and posterior wing margins is unclear.
Expression of
BacA\p35Scer\UAS.cHa in the pupal wing, under the control of
Scer\GAL4sd-SG29.1 results in the loss of TUNEL-positive nuclei at 24h after pupal formation.
Pruning of ddaC dendrites is delayed upon expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4ppk.1.9. The degeneration of dendrites after severing is unaffected however.
The A7 spiracle, which in wild-type males is associated with the A6 tergite, remains posterior to A6 in adult males expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4dsx-GAL4. No A7 tergite develops in these mutants. However, there is excess pleural tissue posterior to the A6 tergite. Furthermore, the A6 tergite of the mutants is significantly narrower than A6 of control males.
Overexpression of
BacA\p35Scer\UAS.cHa in presynaptic neurons under the control of
Scer\GAL4OK6 has no obvious effect on synapse elimination.
Overexpression of
BacA\p35Scer\UAS.cHa in postsynaptic neurons under the control of
Scer\GAL4C57 results in strong arrest of synapse elimination at 7 hours after puparium formation.
Animals expressing
BacA\p35Scer\UAS.cHa in the salivary glands under the control of
Scer\GAL4fkh.PH display persistent condensed salivary gland cell fragments in 94% of pupae, and persistent salivary gland fragments in 6% of pupae.
dsx-SN and
dsx-TN2 cells are protected from programmed cell death in adult females expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4dsx.KI as the number of these neurons is not significantly different from that of wild-type males in these females (these neurons are normally present in both sexes in 48 hour pupae but are specific to males in the adult). Additional neurons are observed in
dsx-pC2 and
dsx-pC3 clusters in females expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4dsx.KI and cells of the TN1 cluster are also present (these cells are never observed in wild-type females). TN1 and TN2 neurons present in these females have contralateral projections that are ordinarily seen only in wild-type males.
Mushroom body neuroblasts are seen in the brains of young adults expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4wor.PA, although their persistence is only transient (mushroom body neuroblasts are not seen in wild-type adults).
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Abd-B-LDN results in a high proportion of males having a half-rotated genitalia (180[o] rotation) phenotype. Time-lapse imaging reveals that the A8a ring (which normally rotates) in the genital disc stays mostly still during the whole process, whereas the A8p ring in the genital disc rotates normally.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4hh-Gal4 results in a high proportion of males having a half-rotated genitalia (180[o] rotation) phenotype.
Expression of
BacA\p35Scer\UAS.cHa simultaneously under the control of both
Scer\GAL4Abd-B-LDN and
Scer\GAL4hh-Gal4 results in 40% of males showing non-rotated genitalia, while the remainder show 90[o] rotation.
Flies expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4GMR.PF show an increase in the size of the eye and also in the size of the ommatidia within the eye compared to control flies.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4109-53 results in an abnormal stalk between egg chambers. The stalk contains more cells than normal and the cells are not properly arranged into a one-cell wide stalk.
At 24 hours after puparium formation, animals expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4fkh.PH still contain salivary gland tissue, in contrast to wild-type animals, where the salivary glands have completely degraded by this stage.
Scer\GAL430A-driven expression of
BacA\p35Scer\UAS.cHa prevents migration of most cells and blocks bonding of the wing surfaces but does not appear to affect delamination from the cuticle. The cells remain within the wing suspended in haemolymph in a round state. In flies heterozygous for
BacA\p35Scer\UAS.cHa, many fewer cells remain within the wing. These cells are round and tightly bound within the wing due to bonding of the wing surfaces; they eventually found brown spots in the wing as the fly ages.
Blocking apoptosis in larval epidermal cells by clonal expression of
BacA\p35Scer\UAS.cHa, under the control of
Scer\GAL4Act5C.PI, results in impaired extrusion of these cells from the epidermis, and failure of haemocyte recruitment and the engulfment process. The few larval epidermal cells that are able to undergo extrusion remain as viable cells under the epithelial layer. Concomitantly, there is a significant decrease in the progression of histoblast nest spreading and in the average number of histoblasts. Some of these animals survive to pharate adults, but survivors show abdominal cuticle clefts with many larval epidermal cells still present.
The number of
Rh5 photoreceptors in the Bolwig's organ at the end of embryogenesis is unchanged relative to wild type in animals expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4so-7.GAL4. However, the number of
Rh6 photoreceptors is dramatically increased to 20-25 instead of the normal number of 8-10.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Tab2-201Y has no effect on the pruning of the axon branches or on the engulfing action of glia in the dorsal lobes of the mushroom body during metamorphosis.
The overall morphology of wing discs expressing
BacA\p35Scer\UAS.cHa in the posterior compartment under the control of
Scer\GAL4hh-Gal4 is normal except for occasional local tissue overgrowth.
Inhibition of caspases in the scutellum, achieved by expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4sca-P309, results in the formation of ectopic macrochaetae on this body part in 39% of cases. Ectopic macrochaetae also appear in 23% of cases when
BacA\p35Scer\UAS.cHa is expressed under the control of
Scer\GAL4dpp.blk1. In contrast, the ectopic expression of
BacA\p35Scer\UAS.cHa in SOP cells, driven by
Scer\GAL4neur-GAL4-A101, does not result in extra macrochaetae. No significant increase in macrochaetae number is seen when
BacA\p35Scer\UAS.cHa is expressed under the control of
Scer\GAL4hs.PB following a heat shock at late third-instar (6-12 h APF), which is during SOP cell formation, or after it (24-30 h APF). At the stage prior to the appearance SOP cells, however,
BacA\p35Scer\UAS.cHa expression leads to extra macrochaetae formation.
X-ray irradiated wing disc clones expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4tub contain a mixture of two types of cell: `live' cells, which have not initiated apoptosis due to the irradiation, and `undead' cells (between 20 and 70% of the clone), which have initiated apoptosis, but have failed to complete programmed cell death. No such cells persist outside the clones in which
BacA\p35Scer\UAS.cHa is expressed. The `live' cells in the X-ray irradiated clones display increased proliferation, while the `undead' cells exhibit an abnormally slow rate of proliferation. X-ray irradiated wing disc clones expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4tub are also associated with local enhancements of growth, with approximately 43% of wing discs exhibiting local deformations and small outpouchings of extra tissues associated with the clones.
Apoptosis is eliminated and extra photoreceptor clusters are formed at the edge of the developing eye in
BacA\p35Scer\UAS.cHa;
Scer\GAL4GMR.PF animals 42 hours after puparium formation. These clusters survive to form ommatidia in adult flies, but are often incomplete.
Males show rotated terminalia and a gap in the dorsal midline.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Vap.P0201 efficiently suppresses the cell death that is normally seen in anterior dMP2 neurons in late stage embryos, such that these neurons are present in late stage embryos.
Flies expressing
BacA\p35Scer\UAS.cHa under the control of one of
Scer\GAL4nub-AC-62,
Scer\GAL4ap-md544,
Scer\GAL4hh-Gal4 or
Scer\GAL4en-e16E have virtually normal wings. Wing vein L5 is slightly shortened in animals expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4hh-Gal4. Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4ap-md544 prevents wing death in the dorsal, but not the ventral, compartment of the wing disc. Animals expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4hh-Gal4 which have been irradiated during the first larval instar show defects in the wing disc; the posterior compartment is abnormal in shape and increased in size. The few adults that do hatch differentiate wings with very abnormal and overgrown posterior compartments. Wing discs of animals expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4hh-Gal4 which have been irradiated during the first or second larval instar show an increase in the ratio of the size of the posterior compartment/anterior compartment compared to control discs and show greater BrdU incorporation in the posterior compartment after irradiation. In addition, cells in the anterior compartment close to the anterior-posterior (AP) border also show an increase in BrdU incorporation. The AP border becomes highly irregular in these irradiated discs and cells of posterior provenance often penetrate into the anterior compartment. These invading posterior cells do not readily mix with anterior ones, but tend to stay in separate groups. Clones in the wing disc expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Act5C.PI which have been induced during the first or second larval instar and then irradiated 24 hours later, show heterogeneous incorporation of BrdU inside the clone. There is often an increase of BrdU levels in the vicinity of these clones.
Expression of
BacA\p35Scer\UAS.cHa, driven by
Scer\GAL4Ddc.PL, results in a statistically significant increase in the number of both serotonergic and dopaminergic neurons after metamorphosis.
When
BacA\p35Scer\UAS.cHa is driven in the wing disc by
Scer\GAL4C10, no effect on the average size of the wing disc is seen. However significantly variability is in the size of the wing disc.
BacA\p35Scer\UAS.cHa;
Scer\GAL4Hsp83.PA males are infertile (males carrying either construct alone are not). Although the initial assembly of the "individualisation complex" at the head of the cyst appears normal, there is a failure of cytoplasm to collect into a normal cystic bulge.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4neur-P72 produces supernumerary polar cells in advanced egg chambers. Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Act5C.PP in clones in the egg chamber results in the production of extra polar cells. No obvious defects are seen in the early development of these egg chambers. 80% of stage 9 egg chambers expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Act5C.PP have round anterior follicle cells, a constant thickness of epithelial cells along the anterior-posterior axis and a homogeneous distribution of anterior follicle cell nuclei (in contrast to wild-type stage 9 egg chambers which have elongated anterior follicle cells, thinning of the epithelium at the anterior end and a reduction in density of follicle cell nuclei at the anterior pole). 91% of stage 10 egg chambers expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4Act5C.PP have defects in delamination and migration of border cells; the border cells are either still stuck at the anterior pole or are only partway between the anterior pole and the oocyte. These clusters often contain supernumerary border cells.
In stage 16 female gonads, where expression of
BacA\p35Scer\UAS.cHa is driven by both
Scer\GAL4how-24B and
Scer\GAL4twi.PB, these gonads appear masculinized; male-specific somatic gonadal precursors persist and join the posterior of female gonads, as in wild-type male embryos. The presence of such cells does not appear to affect ovary formation or oogenesis, since embryos develop into fertile adult females.
Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4neur-P72 suppresses the fragmentation of the glial cell which is normally seen in the thoracic microchaete lineage in the developing notum; at 24 hours after puparium formation (APF), 86% of the clusters contain glial cells in these animals (compared to 8% of clusters in control animals). The ectopically surviving glial cells are tightly associated with axonal processes in 72% of sensory organs studied at 24 hours APF (in 55% of cases the glial cells are located at the growth cone, while in the remaining 17.5% they are associated with the axon), while in 27.5% of the sensory organs studied the glial cells remain within the proximity of the cluster. At 27 hours APF, the axonal network of the notum and its orientation shows no major difference between wild-type pupae and pupae expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4neur-P72. Expression of
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4neur-P72 promotes precocious axonogenesis; axonal growth cones of notal microchaetae are observed 23.5 hours APF in these flies, compared to 25 hours APF in wild type. The cleaning reflex is normal in adults expressing
BacA\p35Scer\UAS.cHa under the control of
Scer\GAL4neur-P72.
In
BacA\p35Scer\UAS.cHa;
Scer\GAL4arm.PS embryos, an ectopic external sensory organ forms near the abdominal ventral multidendritic neuron a1 neuron in many abdominal segments.
When expression is driven by
Scer\GAL4fkh.PH salivary glands survive the cell death that would have been their normal fate. DNA fragmentation is suppressed, though they have progressed to a late stage of autophagy (i.e. vacuoles and associated plasma membranes are often absent from the cytosol).
When expression is driven by
Scer\GAL4cCa.T:Hsim\VP16 in mitotic domain 20 at gastrulation an expansion of the dorsal surface epithelium occurs, though cell death still occurs. Defects are mild compared to those of
Df(3L)H99 embryos.
Scer\GAL4D59-mediated expression causes failure of salivary gland histolysis in 15 hour prepupae, the salivary glands can still be detected 22 hours after puparium formation. The salivary glands at this stage are abnormal in morphology and appear to be partially degraded.
