Project description:Screening for binding partners of the splicing factor SmD3 and changes in interaction upon depletion of the protein Ecdysoneless (Ecd) in the nubbin domain of third-instar larval wing imaginal discs.
Project description:Transcriptomes of Drosophila melanogaster eye-antennal imaginal discs at three sequential larval stages: late 2nd instar (72h after egg laying (AEL)), mid 3rd instar (96h AEL) and late 3rd instar (120h AEL).
Project description:PSC overexpression can cause phenotypes specifically in an rbf1 mutant background, likely due to a sensitization to PSC-induced phenotypes. The goal of this study is to understand the interaction between rbf1 hypomorphic mutation and the overexpression of Polycomb group gene Posterior sex combs. We used Drosophila larval eye imaginal discs that were mutant for rbf1 or overexpressing PSC and compared these to control larval eye discs to assess changes in gene expression. We identified a common set of genes that are deregulated when rbf1 is mutated or when PSC is overexpressed. RNA was extracted from eye imaginal discs dissected from third instar Drosophila larvae. Samples were amplified and hybridized to Affymetrix Drosophila Genome 2.0 Array. To better understand the effects of rbf1 mutation and PSC overexpression, we compared the gene expression of rbf1 mutant eye discs and eye discs overexpressing PSC to control eye discs.
Project description:Gene expression in larval, early third instar eye-antenna discs was assessed to reveal an ATF4 contribution to target gene induction following COX7a knockdown. As hypothesised, these COX7a-RNAi induced target genes require the transcription factor ATF4 for induction, irrespective of concomitant Notch pathway activation through Delta over-expression.
Project description:Third instar larval eye discs provide an in vivo model for cell cycle exit studies. Posterior to the Second Mitotic Wave proliferation is absent in a wild type eye disc. Inactivating mutations in tumor suppressor-like genes can lead to genome wide changes in gene expression that allow for inappropriate bypass of cell cycle exit signals posterior to the Second Mitotic Wave. In a mosaic tissue comprised of two wild type (Canton) populations (distinguished by presence or absence of GFP) there is roughly a 50/50 distribution of both populations in the tissue. Using the same method but comparing Warts (wts) to wild type you see that Warts mutant tissue is ~70-80% of the eye disc and the rest is wild type tissue. In the comparison of rbf1120a v rbf1120a+wts using the same method for mosaic creation we were able to see a similar ~80-85% of the eye disc as rbf1120a+wts. rbf1120a cells have relatively little advantage over wild type cells, but we were able to use entirely homozygous mutant discs for RNA extraction. We used microarrays to detail the global program of gene expression underlying cell cycle exit and identified distinct classes of up-regulated and down-regulated genes during this process. Drosophila third instar larvea were selected and the eye discs from these animals were prepared for RNA extraction and hybridization on Affymetrix microarrays. We used mosaic larval discs but the majority of the tissue was comprised of the mutant allelic combination we desired due to the proliferative advantage of these mutant cells over wild type cells.
Project description:we have performed mRNA-Seq studies of the larval eye and antennal primordial at multiple timepoints within the third larval instar and white prepupal stages
