Graduate school of Agriculture, Meiji University, Kawasaki, Japan.
Anim Sci J. 2014 Feb;85(2):101-11. doi: 10.1111/asj.12101. Epub 2013 Aug 1.
To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation- and proliferation-phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all-trans retinoic acid (ATRA). Quantitative real-time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3-L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty-eight hours after transfection, the messenger RNA expression level of the adipose differentiation-related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)-transfected cells was 3.5% of that in negative control-transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA-transfected cells was 21.8% lower than that in negative control-transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.
为了研究反刍动物肌内脂肪生成相关的基因，我们选择了 16 个具有显著差异表达的基因。这些基因是从我们之前对克隆牛肌内前体脂肪细胞（BIP）系的分化和增殖阶段 SAGE 研究文库中选择的。我们在全反式视黄酸（ATRA）诱导脂肪生成后第 12 天收获 BIP 细胞。定量实时 PCR 证实了之前 SAGE 研究中 15 个基因表达模式的结果。第 6 天，BIP 细胞中 TG 积累显著增加，但在单胃动物对照 3T3-L1 细胞中完全被抑制。ATRA 在第 3 天增强了 6 个基因的表达水平，而在 BIP 细胞的脂肪生成过程中抑制了 8 个基因的表达。转染 48 小时后，其中一个上调基因编码的脂肪分化相关蛋白（ADFP）在 ADFP 小干扰 RNA（siRNA）转染细胞中的信使 RNA 表达水平是阴性对照转染细胞的 3.5%。此外，诱导 6 天后，ADFP siRNA 转染细胞中的 TG 水平比阴性对照转染细胞低 21.8%。对 ATRA 处理后基因表达谱的分析将有助于我们理解牛肌内脂肪生成的分子机制。
