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Identification of G-protein-coupled receptors associated with differentiation and development of porcine subcutaneous preadipocytes.

ABSTRACT: G-protein-coupled receptors (GPCRs) are membrane proteins that enable cells to sense molecular signals and to respond in a broad range of biological processes according to the circumstances of cells. Recent studies elucidated that short-medium chain fatty acids can stimulate GPCRs to activate signaling pathways for adipocyte differentiation. As for differentiation of Porcine Subcutaneous Pre-Adipocyte (PSPA), it has been demonstrated that octanoate supplementation in differentiation medium is essential for growth arrest which triggers differentiation initiation of PSPA followed by lipid synthesis and accumulation to develop into fully matured adipocyte. Previously we detected 10 candidate GPCR genes considered to affect adipocyte differentiation of PSPA. Transfection of small interfering RNA (siRNA) for the candidate genes indicated that knockdown of three genes (A, B and C) obviously decreased the differentiation of PSPA. Therefore we investigated gene expression profiles of PSPA transfected with siRNA for each of three GPCR genes using a porcine custom microarray. Our result indicated that larger numbers of down-regulated genes were detected more than up-regulated genes by the siRNA transfection. Gene ontology analysis elucidated that genes up-regulated by knockdown of Gene A, B and C were categorized in biological processes of skeletal system development, mitosis, and cell adhesion, respectively. Interestingly, knockdown of all three genes commonly down-regulated genes categorized in PPAR signaling pathway and immune response. Differentiation of PSPA might be attributable to the alteration of gene expression profiles regulated by GPCRs identified in this study.

ORGANISM(S): Sus scrofa

PROVIDER: GSE109067 | GEO | 2019/05/31

REPOSITORIES: GEO

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Project description:Adipocytes are dynamic cells that have critical functions to maintain body energy homeostasis. Function of adipocyte is affected by adipogenic differentiation, and exogenous stimulation of biochemical factors such as serotonin and TNF-. Serotonin (5-hydroxytriptamine, 5-HT) is a biogenic monoamine, which known to be involved in lipid metabolism in adipocytes. The TNF- is a proinflammatory cytokines involved in adipocyte biology. Here we investigated the global transcriptome alterations of porcine intramuscular adipocytes at undifferentiated preadipocyte, differentiated adipocytes state, and adipocytes after stimulation with serotonin and TNF- for 12 h by using Agilent’s Porcine (V2) one-color Gene Expression Microarray 4 × 44. Transcriptome analysis revealed that the expression of 443, 261, and 249 genes were altered after differentiation, and after serotonin and TNF- stimulation, respectively. Differentiation process of adipocytes are found to be involved with expression changes of APP, HNF4A, ESR1, EGR1, SRC, HNF1A, FN1, ALB, STAT3, CBL, CEBPB, AR, FOS, CFTR, PAN2, PTPN6, VDR, PPARG, STAT5A and NCOA3 genes which are enriched in the ‘PPAR signaling pathway’ and ‘insulin resistance pathway’. Dose-dependent serotonin stimulation resulted in a decreased fat accumulation in the adipocytes as well as decreased adipocyte proliferation. Serotonin-induced differentially expressed genes in adipocytes were participated in the significant enrichment of GPCR ligand-binding, Cell chemotaxis, blood coagulation and complement pathways, metabolism of lipid and lipoproteins, regulation of lipid metabolism by PPARA; and lipid digestion, mobilization and transport pathways. TNF-α-induced transcriptional modification follows the similar trend, and alteration of many genes shared between both serotonin- and TNF-α stimulations. Our results provide new insights of transcriptome modifications associated with adipogenesis, and exogenous stimulation of serotonin- and TNF- to the porcine intramuscular adipocytes.

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Identification of G-protein-coupled receptors associated with differentiation and development of porcine subcutaneous preadipocytes.

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