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Differential Gene Expression Profile in Pig Adipose Tissue Treated with/without Clenbuterol

ABSTRACT: Background Clenbuterol, a beta-agonist, can dramatically reduce pig adipose accumulation at high dosages. However, it has been banned in pig production because people who eat pig products treated with clenbuterol can be poisoned by the clenbuterol residues. To understand the molecular mechanism for this fat reduction, cDNA microarray, real-time PCR, two-dimensional electrophoresis and mass spectra were used to study the differential gene expression profiles of pig adipose tissues treated with/without clenbuterol. The objective of this research is to identify novel genes and physiological pathways that potentially facilitate clenbuterol induced reduction of adipose accumulation. Results Clenbuterol was found to improve the lean meat percentage about 10 percent (P< 0.5). The adipose cells became smaller and the muscle fibers became thicker with the administration of clenbuterol. The mRNA abundance of 82 genes (ESTs) were found to be statistically differentially expressed based on the Student t-test (P<0.5) in the microarray analyses which containing 3358 genes (ESTs).Theses 82 genes (ESTs) were divided into four groups according to their Gene Ontology Biological Process descriptions. 16 genes were cellular metabolism related genes (including five lipids metabolism related such as apolipoprotein D and apolipoprotein R), 10 were signal transduction related genes, 45 were expressed sequence tags (ESTs) and other 11 others were of various categories. Eleven of the 82 genes (ESTs) were chosen for real-time PCR analysis, with eight of them induced to express similar magnitudes as in the microarray analyses, two could not be detected and one expressed magnitudes that disagreed with the microarray results. Apolipoprotein R was also found to be up-regulated by the proteomic analysis. Conclusions Profound changes were observed in adipose cells after administration of clenbuterol in pigs by histological section and cDNA microarray techniques. The adipose cells became smaller with stimulation by clenbuterol. Changes in mRNA abundance indicated that DNA transcription and protein translation were enhanced in the adipose cells with clenbuterol stimulation. Five lipids metabolism related genes, such as apoD and apoR, were significantly up-regulated. ApoR was up-regulated in both the mRNA transcription and the protein synthesis. ApoR may be one of the critical molecules through which clenbuterol reduces fat accumulation. Keywords: dose response

ORGANISM(S): Sus scrofa

PROVIDER: GSE8093 | GEO | 2007/12/17

SECONDARY ACCESSION(S): PRJNA100943

REPOSITORIES: GEO

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Project description:Background: Clenbuterol, a beta2-adrenergic receptor agonist, is used therapeutically to treat respiratory conditions in the horse. However, by virtue of its mechanism of action it has been suggested that clenbuterol may also have repartitioning affects in horses and as such the potential to affect performance. Clenbuterol decreases the percent fat and increases fat-free mass following high dose administration in combination with intense exercise in horses. In the current study, microarray analysis and real-time PCR were used to study the temporal effects of low and high dose chronic clenbuterol administration on differential gene expression of several skeletal muscle myosin heavy chains, genes involved in lipid metabolism and the β2-adrenergic receptor. The effect of clenbuterol administration on differential gene expression has not been previously reported in the horse, therefore the primary objective of the current study was to describe clenbuterol-induced temporal changes in gene expression following chronic oral administration of clenbuterol at both high and low doses. Steady state clenbuterol concentrations were achieved at approximately 50 hours post administration of the first dose for the low dose regimen and at approximately 18-19 days (10 days post administration of 3.2 μg/kg) for the escalating dosing regimen. Following chronic administration of the low dose (0.8 µg/kg BID) of clenbuterol, a total of 114 genes were differentially expressed, however, none of these changes were found to be significant following FDR adjustment of the p-values. A total of 7,093 genes were differentially expressed with 3,623 genes up regulated and 3,470 genes down regulated following chronic high dose administration. Of the genes selected for further study by real-time PCR, down-regulation of genes encoding myosin heavy chains 2 and 7, steroyl CoA desaturase and the β2-adrenergic receptor were noted. For most genes, expression levels returned towards baseline levels following cessation of drug administration. Conclusion: This study showed no evidence of modified gene expression following chronic low dose administration of clenbuterol to horses. However, following chronic administration of high doses of clenbuterol alterations were noted in transcripts encoding various myosin heavy chains, lipid metabolizing enzymes and the β2-adrenergic receptor.

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Differential Gene Expression Profile in Pig Adipose Tissue Treated with/without Clenbuterol

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