Alterations in skeletal muscle mRNA profile by angiopoietin-1 and angiopoietin-2
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE29508 | GEO | 2012/05/24
SECONDARY ACCESSION(S): PRJNA141455
REPOSITORIES: GEO
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Project description:Analysis of changes in skeletal muscle myoblasts in response to over-expression of angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2). The hypothesis tested in the present study was that over-expression of Ang-1 or Ang-2 will induce the expression of pro-survival and pro-differentiation genes in skeletal myoblasts while inhibiting the expression of pro-apoptotic genes. Results provide important information of how mRNA profile of skeletal myoblasts is altered by Ang-1 and ang-2 and provide first evidence that secondary mediators such as leptin may mediate the enhancement of skeletal myoblasts differentiation by Ang-1 and Ang-2. Total RNA obtained from isolated human skeletal myoblasts after 48hrs of infection with adenoviruses expressing GFP (control), Ang-1 or Ang-2.
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Project description:Neuro-vascular communication is essential to synchronize central nervous system development. Yet, the specific cellular players, defined developmental processes and molecular mechanisms involved in this crosstalk remain poorly characterized. Here we identify the angiopoietin/Tie2 signaling axis as a crucial regulator of dendritic morphogenesis of Purkinje cells (PC) during cerebellum development. We show that in the developing cerebellum Tie2 expression is not restricted to blood vessels but it is also in PCs. Its ligands angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2) are expressed in neural cells and endothelial cells (ECs), respectively. PC-specific deletion of Tie2 results in reduced dendritic arborization, which is recapitulated in neural-specific Ang1-knockout and Ang2 full-knockout mice. Mechanistically, RNA sequencing reveals that Tie2 deficient PCs present alterations in gene expression of multiple genes involved in cytoskeleton organization, dendritic formation, growth and branching. Functionally, those morphological changes are accompanied by alterations in PC network electrophysiology. Altogether, our data propose that the Ang/Tie2 signaling axis serve as mediator of intercellular communication between neural cells, ECs and PCs, required to regulate PC dendritic morphogenesis and function.
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2015-01-01 | E-GEOD-48841 | biostudies-arrayexpress
Project description:Specific Aims ; To identify novel transcriptional events associated with angiogenesis in VEGF and Ang-1 stimulated rat aortic rings. Our studies take advantage of the capacity of rat aortic rings to generate new vessels in collagen gels. Rat aortic rings embedded in collagen gel immediately after excision from the animal produce a self-limited angiogenic response under serum-free conditions and in the absence of exogenous stimuli. This angiogenic response can be dose-dependently promoted by vascular endothelial growth factor (VEGF) and angiopoietin-1 (Ang-1), which are critical regulators of the angiogenic process during embryonal development and postnatal angiogenesis. Aortic rings lose their capacity to spontaneously generate new vessels if embedded in collagen gels 10-14 days after excision. VEGF has the capacity to turn back âonâ these quiescent rings producing florid angiogenesis. Conversely, Ang-1 potentiates an existing angiogenic response, but is unable to turn the quiescent system âonâ. Since VEGF-mediated induction of angiogenic sprouting occurs 1-2 days of treatment, we hypothesize that this process is regulated by a unique set of âangiogenesis inducer genesâ that are activated by VEGF and not by Ang-1. Identification of the proteins encoded by these genes may advance our understanding of the molecular mechanisms that regulate the earliest stages of the angiogenic cascade. Experiment Overall Design: Rat thoracic aortic rings will be excised from 1 month old Fisher 344 rats and placed into suspension culture. 15-20 individual rings will be isolated from each animal and cultured in suspension in serum free medium. The rings will be divided into three groups as described below. (Assay is described in; Nicosia et al., 1997 Am.J.Pathol, 151). Experiment Overall Design: All individual aortic rings isolated from a single animal will be split into three groups, VEGF treated, Ang-1 treated, and untreated controls. Rings dissected from proximal and distal portions of the aorta will be mixed so that positional bias will not be carried through to the RNA preparation. Experiment Overall Design: 13 days post isolation, one set of aortic rings will be treated with 10ng/ml of recombinant human VEGF in serum free EBM media, a second set of rings will be treated with 100ng/ml of human recombinant Ang-1 (R&D Systems 923-AN) in conjunction with 5ug/ml poly-His antibody (R&D Systems MAB050), and a third set of rings will be incubated in serum free medium alone. Experiment Overall Design: All samples were incubated for 18 hours post VEGF and Ang-1 treatment and then RNA was harvested. Experiment Overall Design: With the number of aortic rings typically obtained, we anticipate having 4-6 individual rings from each animal for each experimental condition. Furthermore, 10-100ng of total RNA will be collected per aortic ring, corresponding to 400-600ng of total RNA for each experimental condition per animal. Experiment Overall Design: In order to have a minimum of three independent samples from each experimental condition, three animals were sacrificed, each of which gave rise to three individual RNA samples. The resulting 9 individual RNA samples to be compared with respect to transcriptional profiles. Experiment Overall Design: Total RNA will be extracted from each aortic ring using the Trizol reagent (Invitrogen) protocol. RNA will be stored at â80C until all 9 individual samples have been prepared. Trizol extracted total RNA will be further purified/concentrated using the MicroRNAEasy Kit (Qiagen) with DNAseI digestion.
2007-11-06 | E-GEOD-3355 | biostudies-arrayexpress
Project description:Angiopoietin-Tie2 sytem has been inplicated in both vascular quiescence and angiogenesis. It is unclear how these two opposing signals are regulated from the same receptor-mediated intracellular signal transduction. We have noticed that Tie2 localization upon Angiopoietin stimulation depends upon the presence or absence of cell-cell contacts. Therefore, to identify the downstream signaling of Tie2 upon Angiopoietin stimulation, we performed DNA microarray analyais using RNAs obtained from confluent human umbirical vein endothelial cells (HUVECs) or sparse HUVECs stimulated with after Angiopoietin-1. There is striking difference on gene expression profile between confluent and sparse HUVECs. Keywords: endothelial cell, angiopoietin
2008-04-01 | GSE9677 | GEO
Project description:Angiopoietin-Tie2 sytem has been inplicated in both vascular quiescence and angiogenesis. It is unclear how these two opposing signals are regulated from the same receptor-mediated intracellular signal transduction. We have noticed that Tie2 localization upon Angiopoietin stimulation depends upon the presence or absence of cell-cell contacts. Therefore, to identify the downstream signaling of Tie2 upon Angiopoietin stimulation, we performed DNA microarray analyais using RNAs obtained from confluent human umbirical vein endothelial cells (HUVECs) or sparse HUVECs stimulated with after Angiopoietin-1. There is striking difference on gene expression profile between confluent and sparse HUVECs. Experiment Overall Design: To identify the genes in HUVECs which are upregulated or downregulated by the stimulation with Angiopoietin-1 in the presence (confluent) or absence (sparse) of cell-cell contacts, mRNAs were prepared from either confluent HUVECs or sparse HUVECs stimulated or unstimulated with 100 ng/ml COMP-Ang1 (more potent Angiopoietin-1). RNAs were obtained from HUVECs three times cultured independently three times and mixed as one RNA sample for further DNA microarray analyses. Experiment Overall Design: Microarray (Human Genome U133 Plus 2.0 array, Affymetrix) analyses have been performed in duplicate. Biotin labeled probes were used for the series of analyses. Hybridization and Scan were performed according to the recommendation by Affymetric. Data have been processed following the MAS 5.0 with target intensity = 100.
2008-06-16 | E-GEOD-9677 | biostudies-arrayexpress