In mice, adiponectin receptors (AdipoR) have been found to mediate the effect of adiponectin in muscle and liver in regulation of glucose and fatty acid metabolism. The purposes of this study were to clone these receptors from pig tissues by reverse transcription PCR using mRNA from skeletal muscle and adipose tissue and to investigate the expression of these genes in various pig tissues. Sequences of adiponectin, AdipoR1, and AdipoR2 were determined and found to be highly homologous to those of the human and mouse. The AA sequences predicted for the full-length cDNA of porcine adiponectin, AdipoR1, and AdipoR2 were similar to those of the human and mouse, ranging from 81 to 97% homology, suggesting similar functions of these genes in pigs as in other species. Transcripts for adiponectin were abundant in s.c. adipose tissue in Lee-Sung pigs and in crossbred pigs. Transcripts for AdipoR1 were abundant in heart and skeletal muscle and also detected to a lesser extent (P < 0.05) in adipose tissue, liver, and spleen of the Lee-Sung pigs. Transcripts for AdipoR2 were abundant in s.c. adipose tissue and present to a lesser extent (P < 0.05) in the liver, heart, skeletal muscle, and spleen. These results indicate that the effect of adiponectin may be mediated through these receptors in various porcine tissues. Fasting for 8 h did not have a significant effect on the expression of adiponectin and AdipoR1 mRNA, but it increased (P < 0.05) the AdipoR2 mRNA in the s.c. adipose tissue of crossbred pigs. These results indicate that the AdipoR2-mediated fatty acid oxidation may be responsible at least in part for the fasted state fatty acid oxidation in porcine adipose tissues. The successful cloning of pig adiponectin and adiponectin receptors will enhance the understanding of the involvement of these genes in regulating energy metabolism in pigs.
The purpose of this study was to detect differential expression of genes related to adipocyte differentiation in pigs by suppression subtractive hybridization. Adipocytes and stromal vascular cells (a fraction containing preadipocytes) from pig adipose tissue were isolated for mRNA extraction. The cDNA from preadipocytes was subtracted from the cDNA from adipocytes. The subtracted gene fragments were cloned into pGEM-T Easy TA cloning vector. We selected 384 clones for gene sequence determination and for further analysis. These genes were subjected to a differential screening procedure to confirm the differential expression of genes between the 2 cell types. We found that at least 36 genes were highly expressed in the adipocytes compared with preadipocytes. Among these, 6 genes including 2 novel genes with the greatest differences were selected and confirmed by Northern analysis. We found that angiotensin I-converting enzyme (ACE), ataxia-telangiectasia mutated protein (ATM), calpain 1, and stearoyl coenzyme A desaturase 1 (SCD1) were highly expressed in adipocytes compared with preadipocytes (P < 0.05). The relative mRNA abundance of ACE, ATM, calpain 1, SCD1, and 2 novel genes discovered in the current study was increased at the later stages of adipocyte differentiation (P < 0.05). The results confirmed that the genes involved in lipid metabolism and adipocyte differentiation were highly expressed in porcine adipocytes. However, further investigation is needed to demonstrate specific functions of the novel genes discovered in the current study.
The gene expression of porcine adiponectin and stearoyl coenzyme A desaturase (SCD) was investigated in this study.The partial gene sequences for adiponectin and SCD were amplified by RT-PCR from subcutaneous adipose tissue and cloned by TA cloning techniques. Sequences of these genes were determined and found to be highly homologous to that of other species, suggesting similar function of these genes as in other species. The transcripts of these adipocyte-related genes in pig tissues were measured by Northern analysis. The transcripts for adiponectin and SCD were highly expressed in porcine subcutaneous adipose tissue; the transcripts for SCD were also barely detected in the liver, but the greatest concentrations were in the adipose tissue. In porcine stromalvascular cells (S/V cells) cultured in vitro, transcripts for adiponectin and SCD increased gradually during adipocyte differentiation. The level of adipocyte adiponectin mRNA was associated with late adipocyte differentiation, indicating the gene may not be involved in adipocyte differentiation but has great importance in porcine adipocyte functions. The SCD transcripts were not detectable until 2 d after induction of adipocyte differentiation. It was highly expressed in differentiating porcine adipocytes (2 to 10 d after the induction of adipocyte differentiation), indicating a significant role of SCD in adipocytes.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.