發(fā)布時(shí)間：2018-06-28 06:01

  本文選題：豬 + 單根肌纖維　； 參考：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2017年10期

【摘要】：肌纖維是構(gòu)成肌肉組織的基本結(jié)構(gòu)單位,肉質(zhì)主要受到肌纖維類(lèi)型及其組成的影響。為豐富骨骼肌生長(zhǎng)發(fā)育的體外研究模型,本研究選取剛出生的大白豬(Sus scrofa)后肢肌肉,利用膠原酶消化法分離出單根肌纖維,并采用RT-PCR和免疫熒光染色鑒定不同肌纖維類(lèi)型。對(duì)Ⅰ型膠原酶分離的單根肌纖維進(jìn)行高倍鏡鏡檢,肌纖維表面光滑并且有卵圓形凸起,其為前體骨骼肌衛(wèi)星細(xì)胞,說(shuō)明肌纖維具有良好活性并且其RNA未降解,可進(jìn)行后續(xù)RNA微量提取實(shí)驗(yàn)。逆轉(zhuǎn)錄PCR(reverse transcription PCR,RT-PCR)結(jié)果表明,通過(guò)膠原酶消化法分離趾長(zhǎng)伸肌的8根肌纖維為單根肌纖維,主要類(lèi)型為快速酵解型(Ⅱb);并且通過(guò)慢速氧化型(Ⅰ)抗體對(duì)單根肌纖維進(jìn)行免疫熒光染色驗(yàn)證RT-PCR結(jié)果的可靠性。本研究建立了一種簡(jiǎn)單、低成本的豬骨骼肌單根肌纖維的分離與鑒定方法,為進(jìn)一步研究成熟骨骼肌細(xì)胞發(fā)育過(guò)程及其生物學(xué)功能提供了體外模型。
[Abstract]:Muscle fiber is the basic structure unit of muscle tissue. Meat quality is mainly affected by the type and composition of muscle fiber. In order to enrich the model of skeletal muscle growth and development in vitro, single muscle fiber was isolated from hind limb muscle of newly born large white pig (Sus scrofa) by collagenase digestion, and different types of muscle fibers were identified by RT-PCR and immunofluorescence staining. The single muscle fiber isolated by type I collagenase was examined by high power microscope. The surface of muscle fiber was smooth and oval protuberance. It was a satellite cell of precursor skeletal muscle, which indicated that the muscle fiber had good activity and its RNA was not degraded. The microextraction experiment of RNA can be carried out. The results of reverse transcription-PCR (reverse transcription PCR showed that 8 muscle fibers of extensor digitorum longus were separated into single muscle fibers by collagenase digestion. The main type was rapid fermentative type (鈪，

本文編號(hào)：2077008