Determination of depot-specific mRNA expression by equine adipocytes

Adipose tissue is a complex, highly active metabolic and endocrine organ that is involved in energy homeostasis and regulation of immune function. The purpose of the study was to establish a reliable set of reference genes and to examine mRNA expression in different equine adipose tissue depots. From twelve geldings, euthanased for non research purposes, adipose tissue samples were collected from neck (nuchal), intra-abdominal (mesenterial, right kidney, retroperitoneal) and subcutaneous (loin, tail head) fat depots. Expression level of mRNA of a set of genes was determined in each sample. Reference gene expression stability and optimal number for normalization was evaluated with the geNorm algorithm. The genes ACTB, GAPDH, HPRT, RPL 32 and SDHA were identified as housekeeping genes. Five of the examined genes showed a significant influence of adipose tissue depot on the mRNA expression (P<0.01). Leptin mRNA expression was significantly higher in nuchal adipose tissue when compared to mesenteric adipose tissue. In contrast, CCL5 and IL-10 had lower expression in nuchal adipose tissue. mRNA expression of adiponectin was significantly lower in tail head adipose tissue compared to the nuchal adipose tissue and kidney adipose tissue. For the pro-inflammatory cytokine IL-1β, there was a significant lower expression in the adipose tissue of the loin compared to the adipose tissue depot around the kidneys. No significant effect was found for SOD2 and MMP2 (P>0.05). Expression of inflammation-related genes in equine adipocytes behaves differently between fat depots. Abbreviations: ACTB (actin, beta); GAPDH (glyceraldehyde phosphodehydrogenase); HPRT (hypoxanthine ribosyl transferase), RPL 32 (ribosomal protein 32), SDHA (succinate dehydrogenase complex, subunit A)

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