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猪卫星细胞的表型受到限制,类似于其肌肉起源。

Porcine satellite cells are restricted to a phenotype resembling their muscle origin.

机构信息

Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg 24060.

出版信息

J Anim Sci. 2013 Oct;91(10):4684-91. doi: 10.2527/jas.2012-5804. Epub 2013 Jul 26.

Abstract

Muscles in most domestic animals differ in function and growth potential based largely on muscle fiber type composition. Though much is known about satellite cells (SC), information is limited regarding how populations of SC differ with muscle fiber type, especially in pigs. Therefore, the objective of this study was to isolate and culture SC from red (RST) and white (WST) portions of the semitendinosus muscle of neonatal and adult pigs and determine their capacity to proliferate, differentiate, and express various myosin heavy chain (MyHC) isoforms in vitro. Porcine satellite cells were isolated from RST and WST muscles of 6-wk-old and adult (>6-mo-old) pigs and cultured under standard conditions. Muscle from neonatal pigs yielded nearly 10 times more (P < 0.001) presumptive satellite cells as those from adult pigs, with fusion percentages close to 60% for the former. The RST yielded more (P < 0.001) SC per gram muscle compared to WST, 8.1 ± 0.2 × 10(4) cells versus 6.7 ± 0.1 × 10(4) cells/gram muscle in young pigs, and 9.7 ± 0.4 × 10(3) cells versus 5.5 ± 0.4 × 10(3) cells/gram muscle in adult pigs, respectively. Likewise, satellite cells from RST proliferated faster (P < 0.001) than those from WST across both ages, as indicated by a shorter cell doubling time, 18.6 ± 0.8 h versus 21.3 ± 0.9 h in young pigs, and 23.2 ± 0.7 h versus 26.7 ± 0.9 h in adult pigs, respectively. As a result of shorter times to confluence, satellite cells from RST also formed myotubes earlier than those SC originating from WST. Once induced, however, SC from WST differentiated and fused faster (P < 0.05) as evidenced by fusion percentage within the first 24 h, 41.6% versus 34.3%, respectively; but reached similar ultimate fusion percentages similar to WST by 48 h. Over 90% of MyHC expressed in maximally fused SC cultures from both RST and WST was restricted to the embryonic isoform. Type IIX MyHC mRNA was not detected in any culture. Myotube cultures from RST expressed more (P < 0.01) Type I MyHC isoform mRNA than those from WST, whereas those cultures from WST expressed more (P < 0.05) Type II (including Types IIA and IIB) MyHC transcripts. These data show SC cultures from porcine fast and slow muscles express MyHC profiles largely reflective of their muscle of origin and suggest satellite cells are partially restricted to a particular muscle phenotype in which they are juxtapositioned. Understanding the molecular nature of these intrinsic control mechanisms may lead to improved strategies for augmenting meat animal growth or muscle regeneration.

摘要

大多数家畜的肌肉在功能和生长潜力上存在差异,这主要取决于肌肉纤维类型的组成。尽管人们对卫星细胞(SC)有了很多了解,但关于 SC 群体如何随肌肉纤维类型而变化的信息有限,特别是在猪中。因此,本研究的目的是从新生和成年猪的半腱肌的红(RST)和白(WST)部分分离和培养 SC,并确定它们在体外增殖、分化和表达各种肌球蛋白重链(MyHC)同工型的能力。从 6 周龄和成年(>6 月龄)猪的 RST 和 WST 肌肉中分离出猪卫星细胞,并在标准条件下培养。来自新生猪的肌肉产生的假定卫星细胞数量几乎是成年猪的 10 倍(P<0.001),前者的融合百分比接近 60%。与 WST 相比,RST 每克肌肉产生的 SC 数量更多(P<0.001),年轻猪的 RST 为 8.1±0.2×10(4)个细胞/克肌肉,WST 为 6.7±0.1×10(4)个细胞/克肌肉,成年猪的 RST 为 9.7±0.4×10(3)个细胞/克肌肉,WST 为 5.5±0.4×10(3)个细胞/克肌肉。同样,RST 的卫星细胞增殖速度也快于 WST(P<0.001),这表现在细胞倍增时间较短,年轻猪的 RST 为 18.6±0.8 小时,WST 为 21.3±0.9 小时,成年猪的 RST 为 23.2±0.7 小时,WST 为 26.7±0.9 小时。由于达到汇合的时间更短,因此 RST 的卫星细胞也比源自 WST 的 SC 更早形成肌管。然而,一旦被诱导,WST 的 SC 分化和融合速度更快(P<0.05),这表现在前 24 小时的融合百分比上,分别为 41.6%和 34.3%;但在 48 小时时,两者均达到了与 WST 相似的最终融合百分比。来自 RST 和 WST 的最大融合 SC 培养物中表达的超过 90%的 MyHC 被限制为胚胎同工型。在任何培养物中都未检测到 IIX MyHC mRNA。来自 RST 的肌管培养物表达的 I 型 MyHC 同工型 mRNA 多于 WST,而来自 WST 的肌管培养物表达的 II 型(包括 IIA 和 IIB)MyHC 转录本更多(P<0.01)。这些数据表明,来自猪快肌和慢肌的 SC 培养物表达的 MyHC 谱在很大程度上反映了它们的肌肉来源,并表明卫星细胞部分限于它们所处的特定肌肉表型。了解这些内在控制机制的分子性质可能会导致提高肉类动物生长或肌肉再生的策略得到改善。

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