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:Testes and testes terminal epithelium/seminal vesicles were dissected and separated from 4-5 day old Oregon-R-modENCODE Drosophila melanogaster males. RNA-sequencing was then performed on these two distinct biological structures.
Project description:These arrays were done in the context of a study to probe gene expression variation across Y-chromosome substitution lines of Drosophila melanogaster Keywords: polymorphism, evolution, chromosome substitution
Project description:In order to study the effect of protease treatment on dissected testes, we performed different strategies of cleaning and dissociation on Drosophila melanogaster w1118 larval testes. We collected testes without protease treatment with fatbody just attached around the gonad, fatbody alone dissected from around the testes, testes without fatbody and testes dissociated by either by Papain or Trypsin and Collagenase cocktail. We prepared poly A+ RNA-Seq libraries and performed transcriptional profiling to generate 50 bp stranded single end reads.
