猪骨骼肌单根肌纤维的分离与鉴定方法

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Abstract Meat quality is affected by the type and percent of muscle fiber which is the basic unit of muscle tissue. In order to enrich the in vitro research mode of skeletal muscle growth and development, this study selected the hind-limb extensor digitorum longus of a newborn Landrace piglet (Sus scrofa) as sample, and the single muscle fiber was isolated through collagenase digestion. The type of myofiber was then accurately detected by RT-PCR and immunofluoresence. The single muscle fiber were isolated by collagenase digestion and these muscle fibers had smooth surface and the cells adhered to the myofibers originally, which indicated RNA was qualified for subsequent analysis. The result of RT-PCR indicated that myosin heavy chain Ⅱb (MyHC Ⅱb) mainly expressed in 8 myofibers. Furthermore, the result was confirmed by immunofluorescence with the antibody of myosin heavy chain I (MyHCⅠ). In this study, one simple and cheap detection method for isolation and identification of single muscle fibers of porcine skeletal muscle was established, and the result provides a muscle fiber model, which will be useful for studying the various functions and cellular processes of mature muscles in vitro.

Key words： Pig, Single muscle fiber, Digestion of collagenase, RT-PCR, Immunofluorescence

Received: 20 March 2017 Published: 30 October 2017

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	Articles by authors
	HOU Ye
	HU An
	ZHANG Qiao-Xia
	DU Xiao-Yong
	LI Zhang-Chun
	DIAO Shu-Hong
	YU Hai-Yan

Cite this article:

HOU Ye,HU An,ZHANG Qiao-Xia, et al. Isolation and Identification of Single Skeletal Muscle Fibers in Pigs (Sus scrofa)[J]. , 2017, 25(10): 1714-1720.

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http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2017/V25/I10/1714

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