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大鼠比目鱼肌梭内肌纤维的分离方法

赵雪红, 樊小力, 刘广斌

湖北文理学院医学院机能学部，襄阳 441053；西安交通大学医学院生理教研室，西安 710061；南昆士兰大学生物和物理科学系，图文巴，昆士兰，澳大利亚

摘要

梭囊的存在限制了人们对肌梭功能及其机制的深入研究，本研究旨在探索如何将梭内肌纤维成功地从梭囊中分离出来。应用混合酶液消化的方法，分离大鼠比目鱼肌的梭内肌纤维，显微镜下观察细胞形态，并结合膜片钳技术检测静息膜电位判断梭内肌纤维的功能状态；在多种孵育液中观察、比较梭内肌纤维的活性，寻找适合梭内肌纤维体外生存的液体环境。结果发现，在L-15培养基中孵育消化后，梭内肌纤维的功能状态良好，且能够维持1~2 h；膜片钳技术检测分离出的梭内肌纤维静息膜电位为-45.3±5.1 mV，这与以往在猫及两栖类动物上的在体研究结果相一致，提示分离出的梭内肌纤维功能状态良好，能够满足电生理实验的要求。本研究为进一步研究梭内肌纤维的功能及其机制奠定了良好的方法学基础。

关键词： 肌梭; 梭内肌纤维; 静息膜电位

分类号：R338

[Method of isolation of intrafusal fibres of muscle spindle in soleus of rats.] [Article in Chinese]

ZHAO Xue-Hong, FAN Xiao-Li, LIU Guang-Bin

Medical function Department, Medical College of Hubei University of Arts and Science, Xiangyang 441053, China； Department of Physiology, Medical College of Xi'an Jiaotong University, Xi'an 710061； Department of Biological and Physical Sciences, USQ, Toowoomba, Queensland, Australia

Abstract

Capsule has restricted the further study on muscle spindle function and the involved mechanism. The aim of this study was to explore the isolation method of intrafusal fibres from the isolated rat muscle spindle. Intrafusal fibres were harvested from muscle spindle of soleus muscle in rats treated by neutrase -collagenase digesting. The isolated intrafusal fibres were identified under a microscope，and patch clamp was used to record resting potential. A variety of incubation mediums have been used to find out some survival medium of intrafusal fibers in vitro. The results indicated that the intrafusal fibres could survive for 1~2 h in the Leiboviz`s 15 Medium. The resting potential of intrafusal fibres obtained from patch clamp recording was -45.3±5.1 mV. The results are consistent with others obtained in vivo muscle spindle from cats and frogs. It is suggested that the intrafusal fibres are in good condition and enough to be used in experiments involving patch clamp. This study provides us a new approach in better understanding of muscle spindle activities and the involved mechanism.

Key words: muscle spindle; intrafusal fiber; resting potential

收稿日期：2012-11-20　　录用日期：2013-05-31

通讯作者：樊小力　　E-mail: fanxl@mail.xjtu.edu.cn

引用本文：

赵雪红, 樊小力, 刘广斌. 大鼠比目鱼肌梭内肌纤维的分离方法[J]. 生理学报 2013; 65 (4): 461-465.

ZHAO Xue-Hong, FAN Xiao-Li, LIU Guang-Bin. [Method of isolation of intrafusal fibres of muscle spindle in soleus of rats.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (4): 461-465 (in Chinese with English abstract).