Unknown,Transcriptomics,Genomics,Proteomics

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Genome-wide gene expression analysis of Cirh1a heterozygote state on early embryo in mice

ABSTRACT: CIRHIN is a transcription co-factor, shows postive effect on Hiv-1 LTR enhancer element(NF-κB site); and Cirh1a null is preimplantation lethal, but highly expressed in the early mice embryo (E6.0-E12.5). We hypothesis that Cirh1a's effect on gene transcription of early embryo can be detected with Cirh1a down regulated in Cirh1a+/- state. Result shows that Cirh1a+/- has significant effect on early mice embryo gene transcription, Differentially expressed genes (DEGs) detected are mainly involved in cell proliferation/differenciation, cell cycle progress, embryo development and multi-orgnaogenesis. Littermate embryos (E8.5) were dissected into RNAlater reagent for total RNA extraction, and yolk sac was used for genotyping PCR. Littermate embryos (E9.5) were dissected into RNAlater reagent for total RNA extraction, and yolk sac was used for genotyping PCR. Liver buds from littermate embryos (E11.5) were dissected into RNAlater reagent for total RNA extraction, and yolk sac was used for genotyping PCR. Liver buds from littermate embryos were dissected into RNAlater reagent for total RNA extraction, and yolk sac was used for genotyping PCR.

ORGANISM(S): Mus musculus

SUBMITTER: Andrea Richter

PROVIDER: E-GEOD-51104 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The signaling cascades that direct the morphological differentiation of the vascular system during early embryogenesis are not well defined. To further understand the role of Notch signaling during endothelial differentiation, this study uses both an in vivo gain-of-function and in vivo loss-of-function approach. At embryonic day 9.5, embryos with activated Notch1 signaling in the endothelia display a variety of growth and cardiovascular defects, and die soon after E10.5. Most notably, the extra-embryonic vasculature of the yolk sac displays remodeling differentiation defects. In the wild-type yolk sac, the primary vascular network has begun to reorganize, forming the large primary vessels and the smaller capillaries. In the activated Notch1 embryos remodeling is defective; the vasculature have an enlarged surface with decreased inter-vessel space. Embryos with ablated Notch signaling also display growth and vascular defects at E9.5 similar to the activated Notch1 embryos, however they exhibit a lack of vascular remodeling in the yolk sac, retaining the simple vascular plexus seen at E8.5. These results indicate that Notch signaling plays a critical role in the remodeling of the vasculature in the early embryo, particularly in the extra embryonic region. A conditional transgenic system was used in this study to activate Notch signaling. The ubiquitous ROSA26Notch transgene with a Neo/stop cassette flanked by loxP sites, followed by the N1-ICD cDNA, was recombined with a Tie2-CRE mouse, resulting in the removal of the STOP cassette and the subsequent activation of the Notch1-intracellular domain. This allowed for the overexpression and expansion of Notch signaling in all endothelial cells. Male Tie2-Cre mice were mated with female ROSA26Notch mice and resulting embryos were dissected at embryonic day 9.5. To ablate Notch signaling, Tie2-Cre mice were used in a two generation cross to obtain Tie2-Cre; Rbpj flox/flox embryos. These embryos lack RBPJ binding activity in the endothelia. In both instances embryos were used for genotyping and the yolk sac were separated and used to isolate total RNA with an RNeasy mini kit. The RNA was analyzed with the Mouse Genome 430A Array from Affymetrix. Samples were performed in duplicate, and RNA from wild type yolk sac tissues was compared to activated Notch and RBPJ loss-of-function yolk sac tissues.

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Genome-wide gene expression analysis of Cirh1a heterozygote state on early embryo in mice

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