Project description:The expression of a very large number of genes changes as male germ cells pass through differentiation into spermatids and then sperm. Much of this transcriptional programme requires the activity of the meiotic arrest genes. There are two principle classes, aly-class (including comr) and can-class, here we compare their effects on gene expression We extracted RNA from wild type testes, as well as comr mutants and can mutants, and used microarrays to compare gene expression profiles. We dissected testes from 0-1 day old Drosophila males, taking care to remove accessory glands and ejaculatory ducts. The control sample was red e, the background chromosome on which the can mutant was generated, and had normal testes. The comr mutants were comr[z1340] cn bw. The can mutants were w ; can[3] red e.
Project description:The expression of a very large number of genes changes as male germ cells pass through differentiation into spermatids and then sperm. Much of this transcriptional programme requires the activity of the meiotic arrest genes. There are two principle classes, aly-class (including comr) and can-class, here we compare their effects on gene expression We extracted RNA from wild type testes, as well as comr mutants and can mutants, and used microarrays to compare gene expression profiles.
Project description:Spermiogenesis in Drosophila melanogaster is a highly conserved process and essential for male fertility. In this haploid phase of spermatogenesis, motile sperm are assembled from round cells, flagella are assembled, and needle-shaped nuclei with highly compacted genomes are formed. We aimed at identifying proteins relevant for the maturation phase from spermatids to sperm. As transcription takes place mainly in spermatocytes, and transcripts with relevance for post-meiotic sperm development are translationally repressed for days, we comparatively analysed the prote-ome of larval testes (stages before meiotic divisions), of testes of 1–2-day-old pupae (meiotic and early spermatid stages) and adult flies (late spermatids and sperm). We identified 6677 pro-teins, with 422 solely detected in larval testes, 623 in pupal testes and 634 in adult testes. We analysed a few so far uncharacterized proteins with repect to stage specific expression and im-portance for male fertility. For example, Mst84B (gene CG1988), a very basic cysteine- and lysine-rich nuclear protein, was present in the phase of transition from a histone-based to a pro-tamine-based chromatin structure. CG6332 encodes d-Theg, which is related to the mouse tHEG and human THEG proteins. Mutants of d-Theg lacked sperm in the seminal vesicles and were sterile. The identification of numerous predicted proteins underscores the high potential of pro-teome analysis for future analyses of spermatogenesis.
Project description:The expression of a very large number of genes changes as male germ cells pass through differentiation into spermatids and then sperm. Much of this transcriptional programme requires the activity of the meiotic arrest genes. We extracted RNA from wild type testes, as well as aly mutants and Nxt1 mutants, and used microarrays to compare gene expression profiles. We dissected testes from 0-1 day old Drosophila males, taking care to remove accessory glands and ejaculatory ducts. The control sample expressed a tin-RNAi construct, and had normal testes. The aly mutant were homozygous for aly[5]. The Nxt1 mutants were Nxt1[z2-0488]/Nxt1[DG05102].
Project description:To investigate the mechanisms of gene regulation during Drosophila spermatogenesis, we studied the effects of comr and can mutations on the chromatin of the cells in Drosophila testes by H3K27me3 ChIP-Seq.
Project description:Comr protein was found to be a major regulator of gene activity in drosophila spermatocytes. We obtained Comr binding profile to determine targets of Comr. Comr binding in drosophila male germ line cells was determined using DamID technique. Comparison of Dam-Comr binding to Dam-alone signal in duplicate for each sample type.
