Method of isolation of intrafusal fibres of muscle spindle in soleus of rats / 生理学报

ABSTRACT

Capsule restricts the further study on muscle spindle function and the involved mechanism. The aim of this study was to establish the isolation method of intrafusal fibres from the isolated rat muscle spindle. Intrafusal fibres were harvested from muscle spindle of soleus muscle in rats using neutrase-collagenase digestion. A variety of incubation mediums have been tested to find out an appropriate medium of intrafusal fibers in vitro. Trypan blue staining was used to detect cell death, and patch clamp was used to record resting potential. The results showed that the intrafusal fibres incubated with amine acid-saline solution were almost all dead. DMEM could maintain good condition of the fibres, but excess CO2 ventilation would induce cellular swelling or even death. While Leiboviz's 15 (L-15) medium can guarantee 1-2 h of physiological condition of the intrafusal fibres. Coverslips treated with gelatin, polylysine and serum was the better interfaces for the intrafusal fibres to adhere easily, compared with regularly treated coverslip. The resting potential of intrafusal fibres was (-45.3 ± 5.1) mV, consistent with others obtained from in vivo muscle spindle from cats and frogs. These results suggest that the isolation method of the intrafusal fibres has been successfully established in the present study, providing a new approach in better understanding of muscle spindle activities and the involved mechanism.