Identification of differentially expressed genes related to intramuscular fat development in the early and late fattening stages of hanwoo steers.

Seung-Hwan Lee, Eung-Woo Park, Yong-Min Cho, Sung-Kon Kim, Jun-Heon Lee, Jin-Tae Jeon, Chang-Soo Lee, Seok-Ki Im, Sung-Jong Oh, J M Thompson, Duhak Yoon
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引用次数: 64

Abstract

Marbling of cattle meat is dependent on the coordinated expression of multiple genes. Cattle dramatically increase their intramuscular fat content in the longissimus dorsi muscle between 12 and 27 months of age. We used the annealing control primer (ACP)-differential display RT-PCR method to identify differentially expressed genes (DEGs) that may participate in the development of intramuscular fat between early (12 months old) and late fattening stages (27 months old). Using 20 arbitrary ACP primers, we identified and sequenced 14 DEGs. BLAST searches revealed that expression of the MDH, PI4-K, ferritin, ICER, NID-2, WDNMI, telethonin, filamin, and desmin (DES) genes increased while that of GAPD, COP VII, ACTA1, CamK II, and nebulin decreased during the late fattening stage. The results of functional categorization using the Gene Ontology database for 14 known genes indicated that MDH, GAPD, and COP VII are involved in metabolic pathways such as glycolysis and the TCA cycle, whereas telethonin, filamin, nebulin, desmin, and ACTA1 contribute to the muscle contractile apparatus, and PI4-K, CamK II, and ICER have roles in signal transduction pathways regulated by growth factor or hormones. The final three genes, NID-2, WDNMI, and ferritin, are involved in iron transport and extracellular protein inhibition. The expression patterns were confirmed for seven genes (MDH, PI4-K, ferritin, ICER, nebulin, WDNMI, and telethonin) using real-time PCR. We found that the novel transcription repressor ICER gene was highly expressed in the late fattening stage and during bovine preadipocyte differentiation. This information may be helpful in selecting candidate genes that participate in intramuscular fat development in cattle.

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汉宇阉牛育肥早期和后期肌内脂肪发育相关差异表达基因的鉴定。
牛肉的大理石花纹依赖于多个基因的协调表达。牛在12 ~ 27月龄期间背最长肌肌内脂肪含量显著增加。我们使用退火控制引物(ACP)-差异显示RT-PCR方法来鉴定可能参与早期(12个月大)和后期(27个月大)肥育阶段肌内脂肪发育的差异表达基因(deg)。使用20个任意ACP引物,我们鉴定并测序了14个deg。BLAST搜索结果显示,在育肥后期,MDH、PI4-K、铁蛋白、ICER、NID-2、WDNMI、telethonin、filamin和desmin (DES)基因的表达增加,而GAPD、COP VII、ACTA1、CamK II和nebulin的表达减少。利用Gene Ontology数据库对14个已知基因进行功能分类的结果表明,MDH、GAPD和COP VII参与糖酵解和TCA循环等代谢途径,telethonin、filamin、nebulin、desmin和ACTA1参与肌肉收缩装置,PI4-K、CamK II和ICER参与生长因子或激素调节的信号转导途径。最后三个基因NID-2、WDNMI和铁蛋白参与铁转运和细胞外蛋白抑制。采用实时荧光定量PCR技术确定了7个基因(MDH、PI4-K、铁蛋白、ICER、nebulin、WDNMI和telethonin)的表达模式。我们发现新的转录抑制因子ICER基因在增肥后期和牛前脂肪细胞分化过程中高度表达。这一信息可能有助于选择参与牛肌内脂肪发育的候选基因。
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