Project description:To better understand the signaling and transcriptional events involved in the GDNF-independent emergence of the ureteric bud from the Wolffian duct, microarray expression analysis was performed on embryonic kidneys from wild-type and Ret-deficient mice. Microarray data was used to identify genes and gene networks involved in the GDNF-independent outgrowth of the ureteric bud. Whole embryonic kidneys from E12.5 Ret mutant and wild-type mice were isolated. Isolated kidneys were lysed and RNA was extracted with the Qiagen RNEasy Micro kit. The RNA was amplified using the NuGEn Ovation kit and hybridized to the Affymetrix GeneChip Mouse Whole Genome 430 2.0 microarray. Three biological replicates for Ret-knockout and wild-type kidneys were performed.
Project description:To better understand the signaling and transcriptional events involved in the GDNF-independent emergence of the ureteric bud from the Wolffian duct, microarray expression analysis was performed on embryonic kidneys from wild-type and Ret-deficient mice. Microarray data was used to identify genes and gene networks involved in the GDNF-independent outgrowth of the ureteric bud.
Project description:The goal of this set is to build expression data using FACS isolated Ret-positive cells harvested from a Ret-EGFP reporter mouse (embryonic day E12.5). These can be used as a time point in comparative analysis of ontogeny of genes expressed at various developmental time points.
Project description:For most multigenic disorders, clinical manifestation (penetrance) and presentation (expressivity) are likely to be an outcome of genetic interaction between multiple susceptibility genes. Here, using gene knockouts in mice we evaluated genetic interaction between loss of Ret and loss of Sema3d, two Hirschsprung disease (HSCR) susceptibility genes. We intercrossed Ret and Sema3d double null heterozygotes to generate mice with the nine possible genotypes and assessed survival by counting various genotypes, myenteric plexus development by acetylcholinesterase (AchE) staining and embryonic day 12.5 (E12.5) gut transcriptome by RNA-sequencing. Survival rates of Ret wildtype, null heterozygote and null homozygote mice at E12.5, birth and weaning were not influenced by the genotypes at Sema3d locus and vice-versa. Loss of myenteric plexus was observed only in all Ret null homozygotes, irrespective of the genotypes at Sema3d locus, and Sema3d null heterozygote and homozygote mice had normal gut innervation. As compared to wildtype mice gut gene expression, loss of Ret in null homozygotes led to differential expression of ~300 genes, whereas loss of Sema3d in null homozygotes had no major consequence and there was no evidence supporting major interaction between the two genes influencing gut transcriptome. Overall, given the null alleles and phenotypic assays used, we did not find evidence for genetic interaction between Ret and Sema3d affecting survival, myenteric plexus formation or gut transcriptome.
Project description:The purpose of this experiment was to assess global changes in gene expression pattern in the pancreas from Sox4-/- embryos vs. pancreas from Sox4 wild-type embryos at embryonic day 12.5. [Sox4 gene knock-out (Schilham, M. W. et al. (1996) Defects in cardiac outflow tract formation and pro-B-lymphocyte expansion in mice lacking Sox-4. Nature 380, 711-4.)] Embryonic pancreases from homozygous mutants and their wild-type littermates were harvested. Total RNA was isolated amplified using a linear amplification kit and used to prepare the hybridization samples through indirect labeling. Keywords = Pancreas Sox4 null e12.5 Keywords: parallel sample
Project description:Zinc-finger genes Fezf1 and Fezf2 encode transcriptional repressors. Fezf1 and Fezf2 are expressed in the early neural stem/progenitor cells and control neuronal differentiation in mouse dorsal telencephalon. We compared gene expression profiles of rostral forebrains, which contain the telencephalon and the rostral part of the diencephalon, from embryonic day (E) 9.5, E10.5, and E12.5 wild-type control and Fezf1-/- Fezf2 -/- mouse embryos. The forebrain rostral to the caudal limit of the lateral ventricles was isolated manually from E9.5, E10.5, and E12.5 wild-type and Fezf1-/- Fezf2-/- mice. Total RNAs were isolated by Separsol-RNA I and were used for microarray analyses.
Project description:Purpose: In order to study signaling pathway changes of prenatal chlorpyrifos exposure embryonic kidney development Methods: By using RNA-seq, we studied the transcriptome of E12.5, E14.5, E16.5 and E18.5 prenatal chlorpyrifos exposure embryonic kidneys and DMSO exposure (control) embryonic kidneys Results: We show that Notch signaling pathway and aquaporin family had high expression in chlorpyrifos exposure E18.5 kidney. The nephron progenitor markers had low expression in chlorpyrifos exposure embryonic kidneys
