CX43 KO vs WT cortical astrocytes
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE580 | GEO | 2003/08/06
SECONDARY ACCESSION(S): PRJNA85361
REPOSITORIES: GEO
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Project description:RNA samples extracted from astrocytes cultured from wild type and Cx43 null neonatal mice were dye labeled and individually co-hybridized with a reference of labeled cDNAs pooled from a variety of tissues on eight gene arrays containing 8975 mouse DNA sequences
2010-06-05 | E-GEOD-580 | biostudies-arrayexpress
Project description:Our previous studies on brains and hearts of Cx null mice have revealed that expression level, control and coordination of a very large number of genes are regulated compared to wildtype littermates. This result suggests the possibility that the phenotype of the Cx null animals may include genes not only linked to the intracellular communication that gap junction channels provide but also to genes with very different functions. The question often arises, however, of the degree to which “compensation” occurs in knockouts such that gene regulation depends on pathways altered to make up for the missing gene rather than reflecting normal gene interlinkage. In order to explore this question in the Cx43 null setting, we have compared gene expression patterns in Cx43 null astrocytes with that in wildtype astrocytes acutely treated (36 hrs) with Cx43 siRNA. In these studies, Cx43 levels determined by Western blot analysis were reduced by at least 70% following siRNA treatment, comparable with the 3.24x downregulation for mRNA. For each group of astrocytes, four independent cultures were compared, using oligonucleotide microarrays printed with the mouse Qiagen library. 8060 well annotated unigenes were identifiable in all 8 arrays; of these, 8.2% were upregulated and 6.0% downregulated in Cx43 null astrocytes and 6.2% upreguated and 7.0 downregulated in siRNA treated astrocytes. Interestingly, regulation of 92.3% unigenes significantly regulated in both Cx43 deficient astrocytes had the same orientation, representing a highly significant overlap of gene expression alteration. These experiments thus indicate that the gene regulation in Cx43 null astrocytes is largely due to direct interlinkage rather than to developmental compensation for the missing gene. Keywords: genetic modification
2007-06-20 | GSE8168 | GEO
Project description:Cy3-labeled cDNA from brains of neonatal C57BL Cx43 null, Cx43 heterozygous and Cx32 null mice were compared among themselves and to Cy3-labeled cDNA from brains of neonatal C57BL wildtype mice through Cy5-labeled sample reference prepared at once for the entire experiment from aorta, brain, heart, kidney, liver, lung, ovary/testicles, spleen, and stomach - equal amounts from adult male and female C57BL mice.
2004-11-12 | GSE1954 | GEO
Project description:Cy3-labeled cDNA from brains of neonatal C57BL Cx43 null, Cx43 heterozygous and Cx32 null mice were compared among themselves and to Cy3-labeled cDNA from brains of neonatal C57BL wildtype mice through Cy5-labeled sample reference prepared at once for the entire experiment from aorta, brain, heart, kidney, liver, lung, ovary/testicles, spleen, and stomach - equal amounts from adult male and female C57BL mice.
2010-06-05 | E-GEOD-1954 | biostudies-arrayexpress
Project description:Use of null mutant mice is a powerful way to evaluate the role of specific proteins in brain function. Studies performed on knockout mice have revealed some unexpected roles of the gap junction proteins (the connexins). Thus, analyses of gene expression in connexin43 (Cx43) null brains indicated that deletion of a single gene (Gja1) induced expression level change of numerous other genes located on all chromosomes and involved in a wide diversity of functional pathways. The significant overlap between alterations in gene expression level, control and coordination in Cx43 knockout and knockdown astrocytes raised the possibility that Gja1 represents a transcriptomic node of gene regulatory networks. However, conditional deletion of Gja1 in astrocytes of two mouse strains resulted in remarkably different phenotypes. In order to evaluate the influence of the genetic background on the transcriptome, we performed microarray studies on brains of GFAP-Cre:Cx43f/f C57Bl/6 and 129/SVEV mice. The surprisingly low number of Cx43 core genes (regulated in all Cx43 nulls regardless of strain) and the high number of differently regulated genes in the two Cx43 CKOs indicate high influence of mouse strain on brain transcriptome. The transcriptomes of WT and Cx43 null brains from both C57Bl/6 and SVEV strains were profiled and compared at perinatal and adult time points to learn more about the strain dependence of the Cx43-null phenotype. For this purpose, differently labeled cDNAs from biological replicas (4 of each genotype) were co-hybridized with Duke MO36K mouse oligonucleotide array spotted with 36k Operon oligonucleotides V4.0.
2012-04-13 | E-GEOD-37239 | biostudies-arrayexpress
Project description:BAC array purification of hippocampus astroglial ribosome-bound transcriptome in astrocytes from Aldh1l1:Rpl10a eGFP/ Cx43 KO and Aldh1l1:Rpl10a eGFP/ Cx43FL mice
2019-05-22 | GSE110721 | GEO
Project description:Microarrays were hybridized with a mixture of solutions containing LL2 cDNA labeled with Cy3 or Cy5 (dye-swap) and LL14 cDNA labeled with Cy5 or Cy3 (dye-swap). Keywords: repeat sample
2004-12-29 | GSE2114 | GEO
Project description:We used microarrays to compare the global programme of gene expression in primary cultures of neurons and astrocytes. These data sets were compared to the expression profiles of other tissues, including pancreatic islets, in order to identify a specific neuroendocrine program in pancreatic islets. Neurons and astrocytes were isolated from brain cortex and cultured in vitro for 7 and 20 days respectively. RNA was extracted and hybridized on Affymetrix microarrays. Two biological samples from neurons and astrocytes were analyzed.
2013-07-01 | E-GEOD-42607 | biostudies-arrayexpress
Project description:The main objective of the present report was to unravel the Cx43-interaction network in the heart, and to establish the impact of heart ischemia and I/R upon these interactions. In order to characterize the cardiac Cx43 interactome under ischemia and I/R, a quantitative proteomic analysis was performed, through the combination of immunopurification of endogenous Cx43 (Cx43 IP) and identification of its binding partners with the SWATH-MS approach, using rat hearts maintained in a Langendorff apparatus. In the present approach 444 proteins were identified, including proteins identified based on a single peptide (supplementary table I). From these 444 proteins, 299 (approximately 67 % of the entire dataset) were quantified and compared between the various experimental conditions (including non-specific binding to control IP samples). These 299 proteins were further evaluated by a series of complementary analysis to deciphering the truly interactors of Cx43. According with this evaluation, 236 out of the 299 quantified proteins were considered as putative Cx43 interacting partners in the cardiac context presented The results obtained in this study demonstrate that Cx43 mainly interacts with proteins related with metabolism, signaling and trafficking, and that this interactome can be differentially modulated in diseased hearts. Our results shed new light upon the understanding of Cx43 functions in the heart, both in health and disease, which ultimately may lead to the establishment of new therapeutic targets to modulate cardiac homeostasis.
2015-09-09 | PXD002331 | Pride
Project description:The arrays were hybridized against cDNAs labeled during reverse transcription reactions, which were obtained from bacteria grown under two different conditions (liquid XDM2 and liquid BCYE).
2008-11-27 | GSE13748 | GEO