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Function of an Ultrabithorax minigene in imaginal cells.

An Ultrabithorax (Ubx) minigene constructed from three key Ubx control regions is capable of supporting development of Ubx null mutants throughout larval life and beyond to pharate flies, thereby rescuing the larval lethality due to the homeotic mutation. The cuticle of these flies shows that the minigene provides at least partial Ubx function in each of the four compartments whose morphogenetic pathways are determined by Ubx. We analyse beta-galactosidase patterns in imaginal discs conferred by each individual Ubx control region. From the comparison of these patterns with Ubx expression in Cbx mutants, we infer that long-range repressor elements in the chromosomal Ubx gene play an important role in the generation of Ubx expression patterns in imaginal discs. Expression and function of our Ubx minigenes indicate that Ubx control regions are capable of functioning properly out of context and detached from their normal chromosomal location within the homeotic gene complex.

Animals↗

Positional signaling by hedgehog in Drosophila imaginal disc development.

We describe a dominant gain-of-function allele of the segment polarity gene hedgehog. This mutation causes ectopic expression of hedgehog mRNA in the anterior compartment of wing discs, leading to overgrowth of tissue in the anterior of the wing and partial duplication of distal wing structures. The posterior compartment of the wing is unaffected. Other imaginal derivatives are affected, resulting in duplications of legs and antennae and malformations of eyes. In mutant imaginal wing discs, expression of the decapentaplegic gene, which is implicated in the hedgehog signaling pathway, is also perturbed. The results suggest that hedgehog protein acts in the wing as a signal to instruct neighboring cells to adopt fates appropriate to the region of the wing just anterior to the compartmental boundary.

Alleles↗

Wingless induces transdetermination in developing Drosophila imaginal discs.

Drosophila imaginal discs, the precursors of the adult fly appendages, have been the subject of intensive developmental studies, particularly on cell determination. Cultured disc fragments are recognized not only for the ability to maintain their determined state through extra cell divisions but also for the ability to transdetermine, or switch to the determined state of a different disc. An understanding of transdetermination at a molecular level will provide further insight into the requirements for maintaining cell determination. We find that ectopic expression of the Drosophila gene wingless induces transdetermination of foreleg imaginal disc cells to wing cells. This transdetermination occurs in foreleg discs of developing larvae without disc fragmentation. The in situ-transdetermining cells localize to the dorsal region of the foreleg disc. This wingless-induced transdetermination event is remarkably similar to the leg-to-wing switch that occurs after leg disc culture. Thus we have identified a new approach to a molecular dissection of transdetermination.

Animals↗

Mutations in the 8 kDa dynein light chain gene disrupt sensory axon projections in the Drosophila imaginal CNS.

Mutations in an 8 kDa (8x10(3) Mr) cytoplasmic dynein light chain disrupt sensory axon trajectories in the imaginal nervous system of Drosophila. Weak alleles are behaviorally mutant, female-sterile and exhibit bristle thinning and bristle loss. Null alleles are lethal in late pupal stages and alter neuronal anatomy within the imaginal CNS. We utilized P[Gal4] inserts to examine the axon projections of stretch receptor neurons and an engrailed-lacZ construct to characterize the anatomy of tactile neurons. In mutant animals both types of sensory neurons exhibited altered axon trajectories within the CNS, suggesting a defect in axon pathfinding. However, the alterations in axon trajectory did not prevent these axons from reaching their normal termination regions. In the alleles producing these neuronal phenotypes, expression of the cytoplasmic dynein 8 kDa light chain gene is completely absent. These results demonstrate a new function for the cytoplasmic dynein light chain in the regulation of axonogenesis and may provide a point of entry for studies of the role of cellular motors in growth cone guidance.

Amino Acid Sequence↗

Ectopic expression of wingless in imaginal discs interferes with decapentaplegic expression and alters cell determination.

We have expressed the segment polarity gene wingless (wg) ectopically in imaginal discs to examine its regulation of both ventral patterning and transdetermination. By experimentally manipulating the amount of Wg protein, we show that different thresholds of Wg activity elicit different outcomes, which are mediated by regulation of decapentaplegic (dpp) expression and result in alterations in the expression of homeotic genes. A high level of Wg activity leads to loss of all dorsal pattern elements and the formation of a complete complement of ventral pattern elements on the dorsal side of legs, and is correlated with repression of dpp expression. wg expression in dorsal cells of each disc also leads to dose-dependent transdetermination in those cells in homologous discs such as the labial, antennal and leg, but not in cells of dorsally located discs. When dpp expression is repressed by high levels of Wg, transdetermination does not occur, confirming that dpp participates with wg to induce transdetermination. These and other experiments suggest that dorsal expression of wg alters disc patterning and disc cell determination by modulating the expression of dpp. The dose-dependent effects of wg on dpp expression, ventralization of dorsal cells and transdetermination support a model in which wg functions as a morphogen in imaginal discs.

Animals↗

Ectopic expression of activated Ras1 induces hyperplastic growth and increased cell death in Drosophila imaginal tissues.

The Drosophila Ras1 gene is required for proper cell fate specification throughout development, and the loss-of-function phenotype of Ras1 suggests an additional role in cell proliferation or survival. A direct role for RAS1 in promoting cell proliferation, however, has not been established. We show that expression of an activated form of RAS1 (RAS1V12) during Drosophila imaginal disc development is sufficient to drive ectopic cell proliferation and hyperplastic tissue growth. In addition, expression of RAS1V12 induces widespread cell death in the imaginal discs, including cells not expressing the transgene, which results in ablation of adult structures. Loss-of-function mutations in the genes encoding RAF, MEK, MAPK and KSR dominantly suppress RAS1V12-induced cell proliferation. Furthermore, two RAS effector loop mutations (E37G and Y40C) that block the RAS-RAF interaction, also suppress RAS1V12-induced proliferation, consistent with a requirement for the MAPK cascade during the RAS1 mitogenic response. These two RAS effector loop mutants, however, retain some activity and can act synergistically with a MAPK gain-of-function mutation, suggesting that RAS1 may also act through signaling pathway(s) distinct from the MAPK cascade.

Animals↗

double-time is identical to discs overgrown, which is required for cell survival, proliferation and growth arrest in Drosophila imaginal discs.

We have isolated the discs overgrown gene of Drosophila and shown that it encodes a homolog of the Casein kinase I(delta)/(epsilon) subfamily and is identical to the double-time gene. However, in contrast to the weak double-time alleles, which appear to affect only the circadian rhythm, discs overgrown alleles, including bona fide null alleles, show strong effects on cell survival and growth control in imaginal discs. Analysis of their phenotypes and molecular lesions suggests that the Discs overgrown protein is a crucial component in the mechanism that links cell survival during proliferation to growth arrest in imaginal discs. This work provides the first analysis in a multicellular organism of Casein kinase I(delta)/(epsilon) functions necessary for survival. Since the amino acid sequences and three-dimensional structures of Casein kinase I(delta)/(epsilon) enzymes are highly conserved, the results suggest that these proteins may also function in controlling cell growth and survival in other organisms.

Amino Acid Sequence↗

Hedgehog is required for activation of engrailed during regeneration of fragmented Drosophila imaginal discs.

Surgically fragmented Drosophila appendage primordia (imaginal discs) engage in wound healing and pattern regulation during short periods of in vivo culture. Prothoracic leg disc fragments possess exceptional regulative capacity, highlighted by the ability of anterior cells to convert to posterior identity and establish a novel posterior compartment. This anterior/posterior conversion violates developmental lineage restrictions essential for normal growth and patterning of the disc, and thus provides an ideal model for understanding how cells change fate during epimorphic pattern regulation. Here we present evidence that the secreted signal encoded by hedgehog directs anterior/posterior conversion by activating the posterior-specific transcription factor engrailed in regulating anterior cells. In the absence of hedgehog activity, prothoracic leg disc fragments fail to undergo anterior/posterior conversion, but can still regenerate missing anterior pattern elements. We suggest that hedgehog-independent regeneration within the anterior compartment (termed integration) is mediated by the positional cues encoded by wingless and decapentaplegic. Taken together, our results provide a novel mechanistic interpretation of imaginal disc pattern regulation and permit speculation that similar mechanisms could govern appendage regeneration in other organisms.

Animals↗

A targeted gene silencing technique shows that Drosophila myosin VI is required for egg chamber and imaginal disc morphogenesis.

We report that Drosophila unconventional myosin VI, encoded by Myosin heavy chain at 95F (Mhc95F), is required for both imaginal disc and egg chamber morphogenesis. During oogenesis, Mhc95F is expressed in migrating follicle cells, including the border cells, which migrate between the nurse cells to lie at the anterior of the oocyte; the columnar cells that migrate over the oocyte; the centripetal cells that migrate between the oocyte and nurse cells; and the dorsal-anterior follicle cells, which migrate to secrete the chorionic appendages. Its function during development has been studied using a targeted gene silencing technique, combining the Gal4-UAS targeted expression system and the antisense RNA technique. Antibody staining shows that the expression of myosin 95F is greatly decreased in follicle cells when antisense Mhc95F RNA is expressed. Interfering with expression of Drosophila myosin VI at various developmental stages frequently results in lethality. During metamorphosis it results in adult flies with malformed legs and wings, indicating that myosin VI is essential for imaginal disc morphogenesis. During oogenesis, abnormal follicle cell shapes and aberrant follicle cell migrations are observed when antisense Mhc95F is expressed in follicle cells during stages 9 to 10, suggesting that the Drosophila myosin VI is required for follicle cell epithelial morphogenesis.

Animals↗

Developing practice through the professional imagination.

How can the perspective of the service user be made real and helpful in the professional education of community nurses? This article presents one answer to this question through encouraging nurses to write fictional accounts of how their practice appears to their own service users. Arising from a research project on the "patchwork text" assignment format in a professional degree course at Anglia Polytechnic University, the article presents two such accounts by community nurses undertaking the course. Their accounts illustrate the insights into professional issues that are produced by an explicitly imaginative exercise. A particularly important outcome, not expected by the researchers, was that the commentaries written by the nurses on their imagined scenarios, led, not to a discussion of "issues" (as in many cases) but to a precise and practical critique of their actual practice, suggesting a further dimension of the educational value of writing fiction as part of professional education courses.

Attitude of Health Personnel↗

Subjective improvement in PTSD patients with treatment by imaginal exposure or cognitive therapy: session by session changes.

A self-rating scale, the Subjective Symptom Checklist (SSC) was developed to assess PTSD patients' perception of their symptoms between treatment sessions. Reliability and validity of the scale were acceptable. The scale was used in a treatment trial comparing the efficacy of cognitive therapy versus imaginal exposure. Conventional pre-post treatment comparisons using standardized assessments showed no differences between the two treatments. However, when patients who failed to respond to treatment were excluded, group treatment effects became apparent. Patients who received imaginal exposure showed a significantly greater reduction in subjective ratings of their symptoms than did those who received cognitive therapy. It was cautiously concluded that although some PTSD patients could not tolerate exposure, those who could may receive greater subjective benefit than those who received cognitive therapy.

Adult↗

six-banded, a novel Drosophila gene, is expressed in 6 segmental stripes during embryonic development and in the eye imaginal disc.

We have characterised a Drosophila P-element enhancer detector insertion F125, which is expressed in the embryonic head and CNS as well as in various third instar imaginal discs. In an attempt to identify the gene with the equivalent expression pattern, we have characterised an adjacent gene. It encodes two novel conceptual proteins: Type I (1182 amino acids) and Type II, representing a shorter form of 774 amino acids truncated at both termini relative to Type I that is generated by alternative splicing. Based on its embryonic expression pattern, the gene was called six-banded (sba). Both splice forms are expressed in a unique embryonic pattern: initially as 6, then 12 stripes during early stages of embryonic development. Subsequently, expression is found in the developing trachae and during larval development is restricted to the eye imaginal disc where both transcripts are present immediately anterior to and behind the morphogenetic furrow. While sba expression in the eye antennal disc is mirrored by the expression of the adjacent F125 P-element, other patterns reported by this enhancer detector are not mimicked by the sba gene suggesting that the expression of the P-element represents a 'composite' of the effects of nearby enhancers.

Animals↗

[Study of temperature-sensitive mutations in the virilis group of Drosophila. 4. Phenogenetics of a temperature-sensitive mutation affecting the development of imaginal discs in Drosophila virilis Sturt].

The phenogenetics of a new temperature-sensitive mutation diskless-ts (dl-ts, 2nd chromosome) has been studied in D. virilis. The rearing of larvae from the 1st instar at the temperature 31 degrees resulted in the complete or partial arrest of the development of imaginal discs and, consequently, the death of larvae prior to the pupation, at the prepupa stage. The change of temperature from 25 to 31 degrees during the second half of development in the 3rd instar affects the differentiation of imaginal discs. Some organs differentiate completely or partially (eyes, wings, legs) whereas the rest do not develop at all. The sensitive period embraces the whole larval development till the beginning of pupation.

Animals↗

[Radiosensitivity of somatic cells and imaginal disks of male and female Drosohila].

Primary chromosome damages as well as the frequency of spontaneous and X-rays induced chromosome aberrations in Drosophila melanogaster males and females are studied. It is found using cytofluorimetric method that primary chromosome damages in ganglia cells of females and males are the same. In these cells as well as in cells of imaginal discs the frequency of induced chromosome aberrations, except gaps, is considerably higher for females. Ganglia cells of females and males of Drosophila are found not to differ from each other in their proliferation activity. The frequency of morphoses for both sexes is also the same. The assumption is made concerning the role of the non-identical repair in the increased mutability of female somatic cells, which is more intensive in this sex, as well as concerning more intensive identical repair in imaginal discs of females.

Animals↗

Measures of tinnitus: step size, matches to imagined tones, and masking patterns.

Measures of the pitch (0.2% step) and loudness (2 dB step) of tinnitus were obtained using a forced-choice double-staircase procedure. In the first experiment, for seven subjects with tinnitus, the difference limen of pitch and loudness matches made to tinnitus within a single block of trials was not significantly different from that of the same measurements made to comparable external stimuli. In the second experiment, subjects matched the pitch of real and imagined tones. For 3 of 7 subjects with tinnitus and for 2 of 14 normal-hearing subjects, the difference limens were nearly equal. In the first experiment, therefore, three of the subjects with tinnitus might have matched to an imagined tone and not to their tinnitus. In the third experiment, thresholds of tones near the presumed tinnitus frequency were measured. Thresholds were approximately equal in the presence of the tinnitus. However, in the presence of a masking tone at the presumed tinnitus frequency and level, thresholds were unequal. Thus, tinnitus did not behave as a tonal masker.

Acoustic Stimulation↗

[A new allele variant of ssa and its participation in regulating the proliferation of the stem elements of the leg and antenna imaginal disks in Drosophila melanogaster].

Using a highly mutable FM/w oc system, we have established a new allele of ssa40aSc mutation whose phenotype is identical to ssa40aNs compound described in the literature. The following features are characteristic of the ssa40aSc flies phenotype: (1) the increased number of sex comb teeth, (2) complete fusion of tarsal segments, and (3) the decreased bristle size corresponding to that of ss flies. The first two features are evidence for the impaired stem cell proliferation in the antennal and leg imaginal discs which are determined to form distal structures. This assumption was experimentally confirmed when we transplanted leg imaginal discs from III instar larvae of different age into prepupae. The observed phenomenon is probably due to the defects of the Antp and Pc translation products binding site in the ss locus.

Alleles↗

Pattern formation in imaginal discs.

Imaginal discs are sacs of folded epithelium arising during embryogenesis. They proliferate during the larval instars, and at metamorphosis secrete the adult cuticle, therefore being responsible for the characteristic surface patterning of this insect. Each disc has intrinsic growth controls and their cell lineage shows constraints known as compartments. Spatial patterns emerge through interaction between adjacent cells. Molecular genetic analysis of mutants with changed pattern has implicated transcription factors, secreted, membrane-bound and growth factor related proteins in the position-signalling mechanism. Their accessibility to contemporary cell biological techniques makes imaginal discs a model system for investigating patterning in animal tissues.

Animals↗

Cuticle secretion in Drosophila wing imaginal discs in vitro: parameters of exposure to 20-hydroxy ecdysone.

We have cultured Drosophila wing imaginal discs in vitro under a variety of hormonal conditions in order to determine whether cuticle secretion is enhanced by a withdrawal of 20-hydroxy ecdysone at one of two points in development, corresponding to the drop in hormone titer during the prepupal period, and to the fall in hormone levels during the later stages if imaginal differentiation. We found that these treatments did not enhance either pupal or adult cuticle secretion.

Animals↗