The expression of genes related to adipocytes in pigs

The purpose of this study was to detect differential expression of genes related to adipocytes in pigs by suppression subtractive hybridization (SSH). 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 two cell types in pigs. We found that at least 36 genes were highly expressed in the adipocytes compared with preadipocytes. Among these, six genes including two 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), calpain1, 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, calpain1, SCD1, and two 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.目錄----------------------------------------------------I 表次----------------------------------------------------II 圖次----------------------------------------------------III I、摘要--------------------------------------------------1 II、文獻檢討---------------------------------------------2 III、材料與方法-----------------------------------------27 IV、結果------------------------------------------------33 V、討論-------------------------------------------------39 VI、結論------------------------------------------------42 VII、參考文獻-------------------------------------------4