Unknown,Transcriptomics,Genomics,Proteomics

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Dissecting Stages of the Notch signaling pathway during Kidney Development by Oligonucleotide Array Sequence Analysis

ABSTRACT: Samples E12/E13/E14/E16/E18: We aims to screen out different gene expression profile in Embryo kidney on different gestation stages of the Notch signaling pathway Results from the various study components can help to screening important candidate genes during embryonic kidney development. Keywords: Embryo kidney, Development, Notch signaling pathway, Oligonucleotide Array Sequence Analysis The Oligo GEArray Assay comprises various components: RNA isolation,Assesing RNA yield and quality,cRNA labeling and synthesis,Hybridization,Chemiluminescent detection and Image acquisition and data analysis. Samples E12: This study has been accomplished with Embryo kidney on gestation 12, 3 techinical replicates. Samples E13: This study has been accomplished with Embryo kidney on gestation 13, 3 techinical replicates. Samples E14: This study has been accomplished with Embryo kidney on gestation 14, 2 techinical replicates. Samples E16: This study has been accomplished with 1Embryo kidney on gestation 16, 2 techinical replicates. Samples E18: This study has been accomplished with Embryo kidney on gestation 18, 2 techinical replicates.

ORGANISM(S): Mus musculus

SUBMITTER: Jiang Hong

PROVIDER: E-GEOD-28496 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The hepatocyte growth factor (HGF)/c-Met signaling pathway is known to mediate vascularization. We have previously demonstrated that expression of a human HGF transgene in the small airways produced mice (HGF TG) that were more susceptible to the tobacco carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). We also have observed that HGF TG mice display significantly enhanced vascularization in the lungs that increases over time compared to wild-type (WT) littermates. To analyze which genes might contribute to increased vascularization from HGF overexpression in the airways, RNA and protein were isolated from whole lungs of individual HGF TG and WT adult mice. We profiled the mRNA expression of several hundred genes representative of six biological pathways involved in transformation, angiogenesis, and tumorigenesis using two commercial microarrays. Significant changes in expression over a 1.5-fold boundary were also observed in lung tumors derived from NNK-treated HGF TG mice. Lung tumors were induced by exposing mice to four weekly i.p. injections of 3mg NNK (15μg/μl) over 2 weeks. Whole lungs from control untreated animals were dissected after sacrifice at 10, 20, or 40 wks of age, and NNK induced lung tumors were dissected from the animals at 20 or 40 weeks of age. Total RNA was extracted from whole lung or isolated tumors from HGF TG or WT mice using TRIzol reagent and the Array Grade Total RNA Isolation Kit. The cDNA was generated and labeled using the TrueLabeling-AMP Linear RNA Amplication Kit. RNA was analyzed from 28 mice in total. The angiogenesis array was used to analyze 10 samples taken from 40 week old mice (HGF TG untreated [n=4], WT untreated [n=4], HGF TG NNK treated [n=1], WT NNK treated [n=1]) and 6 samples from 20 week old mice (HGF TG untreated [n=2], WT untreated [n=2], HGF TG NNK treated [n=1], WT NNK treated [n=1]). The cancer gene array was used to analyze 8 samples taken from 40 week old mice (HGF TG untreated [n=2], WT untreated [n=2], HGF TG NNK treated [n=2], WT NNK treated [n=2]) and 4 samples from 20 week old mice (HGF TG untreated [n=2], WT untreated [n=2]).

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Dissecting Stages of the Notch signaling pathway during Kidney Development by Oligonucleotide Array Sequence Analysis

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