Proliferation Of Imaginal Cells Research Articles

TheDrosophila melanogasterseptin geneSep2has a redundant function with the retrogeneSep5in imaginal cell proliferation but is essential for oogenesis

Septins are cytoskeletal proteins that form hetero-oligomeric complexes and function in many biological processes, including cytokinesis. Drosophila melanogaster has five septin genes. Sep5, which is the most recently evolved septin gene in Drosophila, is a retrogene copy of Sep2. Sep5 mutants appear wild type, whereas Sep2 mutant females are semisterile. Their ovaries have egg chambers containing abnormal numbers of nurse cells. The egg chamber phenotype is rescued to wild type by expressing a Sep2 cDNA, but it is only partially rescued by expressing a Sep5 cDNA, showing that these paralogs have diverged in function at the protein level. Sep2 Sep5 double mutants have an early pupal lethal phenotype and lack imaginal discs, suggesting that these genes have redundant functions during imaginal cell proliferation.

The 20-hydroxyecdysone-induced signalling pathway in G2/M arrest of Plodia interpunctella imaginal wing cells

The mechanisms involved in the control of cellular proliferation by the steroid hormone 20-hydroxyecdysone (20E) in insects are not known. We dissected the 20E signalling pathway responsible for G2/M arrest of imaginal cells from the IAL-PID2 cells of the Indian meal moth Plodia interpunctella. We first used a 5′–3′ RACE-based strategy to clone a 4479bp cDNA encoding a putative P. interpunctella HR3 transcription factor named PiHR3. The deduced amino acid sequence of PiHR3 was highly similar to those of HR3 proteins from other lepidopterans, e.g. Manduca sexta and Bombyx mori. Using double-stranded RNA-mediated interference (dsRNAi), we then succeeded in blocking the ability of 20E to induce the expression of PiEcR-B1, PiUSP-2 and PiHR3 genes that encode the P. interpunctella ecdysone receptor B1-isoform, Ultraspiracle-2 isoform, the insect homologue of the vertebrate retinoid X receptor, and the HR3 transcription factor. We showed that inhibiting the 20E induction of PiEcR-B1, PiUSP-2 and PiHR3 mRNAs prevented the decreased expression of B cyclin and consequently the G2/M arrest of IAL-PID2 cells. Using this functional approach, we revealed the participation of EcR, USP and HR3 in a 20E signalling pathway that controls the proliferation of imaginal cells by regulating the expression of B cyclin.

Stage- and cell-specific expression of ecdysone receptors and ecdysone-induced transcription factors during midgut remodeling in the yellow fever mosquito, Aedes aegypti

In insects, especially in mosquitoes that are adult blood feeders, midgut remodeling is an important event during metamorphosis. It involves two processes viz., programmed cell death (PCD) of larval cells, and proliferation and differentiation of imaginal cells to form pupal/adult midgut. These processes are regulated by 20-hydroxyecdysone (20E) and juvenile hormone (JH), but the signaling mechanisms, which trigger specific changes remain poorly understood. Here, we report stage- and cell-specific expression of ecydone receptor (EcR), ultraspiracle (USP), broad (Br), E75B and hormone receptor 3 (HR3) during midgut remodeling in Aedes aegypti. In Ae. aegypti both EcR and USP genes code for two isoforms each and the expression of mRNA for these isoforms showed both stage- and cell-specific regulation. In general, EcR-B and USP-A mRNAs were detected during larval stages in larval cells, and EcR-A and USP-B mRNAs were detected during pupal stages in imaginal cells. These data suggest that EcR-B/USP-A heterodimer is important for PCD of larval cells and EcR-A/USP-B heterodimer is important for formation of pupal/adult midgut. Broad Z1 mRNA was detected only in the larval cells suggesting its primary role in PCD. It is likely that E75B and HR3 are probably involved in both PCD and imaginal cell proliferation and differentiation as their mRNAs were expressed in the larval as well as in imaginal cells. Application of JH analog, methoprene, lowered or delayed the expression of all the genes studied. These data suggest that 20E plays a major role in midgut remodeling and coordinates this process through stage- and cell-specific expression of different isoforms of nuclear receptors and transcription factors in the target larval and imaginal cells.

The protein 4.1, ezrin, radixin, moesin (FERM) domain of Drosophila Coracle, a cytoplasmic component of the septate junction, provides functions essential for embryonic development and imaginal cell proliferation.

Coracle is a member of the Protein 4.1 superfamily of proteins, whose members include Protein 4.1, the Neurofibromatosis 2 tumor suppressor Merlin, Expanded, the ERM proteins, protein tyrosine phosphatases, and unconventional myosins. Recent evidence suggests that members of this family participate in cell signaling events, including those that regulate cell proliferation and the cytoskeleton. Previously, we demonstrated that Coracle protein is localized to the septate junction in epithelial cells and is required for septate junction integrity. Loss of coracle function leads to defects in embryonic development, including failure in dorsal closure, and to proliferation defects. In addition, we determined that the N-terminal 383 amino acids define an essential functional domain possessing membrane-organizing properties. Here we investigate the full range of functions provided by this highly conserved domain and find that it is sufficient to rescue all embryonic defects associated with loss of coracle function. In addition, this domain is sufficient to rescue the reduced cell proliferation defect in imaginal discs, although it is incapable of rescuing null mutants to the adult stage. This result suggests the presence of a second functional domain within Coracle, a notion supported by molecular characterization of a series of coracle alleles.

Minidiscs encodes a putative amino acid transporter subunit required non-autonomously for imaginal cell proliferation

Drosophila minidiscs mutant larvae have smaller imaginal discs than wild-type larvae. However, transplantation experiments have revealed that minidiscs mutant imaginal discs can grow if cultured in non-mutant hosts. These data suggest that minidiscs is required in one or more non-imaginal tissues for synthesis and/or secretion of a diffusible factor that stimulates imaginal cell proliferation. The 2.3 kb minidiscs transcript accumulates in the larval fat body and encodes a protein containing 12 putative membrane spanning domains that is similar in sequence to amino acid transporter subunits from other eukaryotes, including humans. We propose that in response to amino acid uptake by the transporter encoded by minidiscs, the fat body secretes a diffusible factor required for imaginal disc proliferation.

A silencer is required for maintenance of transcriptional repression throughout Drosophila development.

Transcriptional silencing by the Polycomb Group of genes maintains the position-specific repression of homeotic genes throughout Drosophila development. The Polycomb Group of genes characterized to date encode chromatin-associated proteins that have been suggested to form heterochromatin-like structures. By studying the expression of reporter genes, we have identified a 725 bp fragment, called MCP725, in the homeotic gene Abdominal-B, that accurately maintains position-specific silencing during proliferation of imaginal cells. Silencing by MCP725 requires the Polycomb and the Polycomblike genes, indicating that it contains a Polycomb response element To investigate the mechanisms of transcriptional silencing by MCP725, we have studied its temporal requirements by removing MCP725 from the transgene at various times during development. We have discovered that excision of MCP725 during larval stages leads to loss of silencing. Our findings indicate that the silencer is required for the maintenance of the repressed state throughout cell proliferation. They also suggest that propagation of the silenced state does not occur merely by templating of a heterochromatin structure by virtue of protein-protein interactions. Rather, they suggest that silencers play an active role in the maintenance of the position-specific repression throughout development.

Genetic analysis of the Drosophila cdc2 homolog

We have identified mutations in the Drosophila cdc2 gene. The recessive lethality of these mutant alleles was rescued after P-element-mediated transformation with a genomic cdc2 fragment. Sequence analysis of amorphic alleles revealed non-conservative exchanges in evolutionary conserved positions. These alleles caused lethality at the larval-pupal interphase due to the absence of imaginal tissues. Embryonic lethality resulted when the maternal Dm cdc2 contribution was reduced through the use of a temperature-sensitive allele. Dm cdc2 function, therefore, is essential for cell proliferation throughout development. Dm cdc2 function is clearly required for mitosis, but no evidence for a requirement in S-phase was obtained. The reversible block of the mitotic proliferation which was observed in the PNS of mutant embryos occurred exclusively in the G2-phase. Moreover, while the mitotic proliferation of imaginal cells was blocked in the amorphic mutant larvae, non-imaginal larval cells continued to grow and endoreplicate their DNA. The Dm cdc2 mutant phenotype could neither be rescued with Dm cdc2c (encoding a cdc2-like kinase) nor enhanced by a reduction of the Dm cdc2c gene dose. These results indicate that the Dm cdc2- and Dm cdc2c-kinases control different processes.

One of the protein phosphatase 1 isoenzymes in Drosophila is essential for mitosis

Drosophila has four loci encoding type 1 protein serine/threonine phosphatases (PP1s). Here we describe mutations in one of these genes, at 87B on chromosome 3. Mutants die at the larval-pupal boundary with little or no imaginal cell proliferation. Neuroblasts are delayed in progress through mitosis and show defective spindle organization, abnormal sister chromatid segregation, hyperploidy, and excessive chromosome condensation. Germline transformation of mutant files with the wild-type PP1 87B gene restores normal mitosis, viability, and fertility. These results show that PP1 activity is required for mitotic progression and that the other loci cannot supply sufficient activity to complement loss of expression of the PP1 87B gene. Alternatively, the PP1 87B product may have a distinct specialized function in mitosis.

Genetic interactions between the Polycomb locus and the Antennapedia and Bithorax complexes of Drosophila.

We have studied the embryonic and adult phenotypes of genetic combinations between Polycomb (Pc), Regulator of bithorax (Rg-bx) and the genes of the Bithorax complex (BX-C) and the Antennapedia complex (ANT-C). The products of Pc and Rg-bx genes act antagonistically, their mutant combinations leading to the ectopic expression of genes of the BX-C and ANT-C. The genetic analysis of the Pc locus suggests it is a complex gene. Pc+ products behave as members of a "regulatory set" that negatively control the expression of BX-C and ANT-C genes. Genetic combinations between different doses of Pc, Rg-bx and the genes of the BX-C and ANT-C have phenotypes which may be interpreted as resulting from ectopic derepression of posterior selector genes repressing selector genes of anterior segments. The transformation phenotypes of certain genetic combinations differ in embryos and adults. A model of regulation of the BX-C and the ANT-C genes during the imaginal cell proliferation is presented, in which the specification state is maintained by self-activation of a given selector gene and down modulation of other selector genes in the same cell.

A clonal analysis of the requirement for the trithorax gene in the diversification of segments in Drosophila

ABSTRACT The requirement for the trithorax gene during imaginal cell proliferation has been analysed by clonal analysis. Clones carrying the allelic Regulator of bithorax and trithorax mutations were induced by mitotic recombination at different developmental stages. The results indicate that the trithorax gene is required at least until the beginning of the third larval instar to ensure the correct differentiation of head, thoracic, and abdominal structures, and are consistent with a role for the gene in maintaining the appropriate levels of Antennapedia and bithorax complex expression.