Expression of myosin heavy chain isoforms in regenerated muscle spindle fibres after muscle grafting in young and adult rats – plasticity of intrafusal satellite cells

Abstract

Satellite cells are the myogenic precursor cells of postnatal skeletal muscles. After muscle injury they can proliferate, differentiate, fuse and form myofibres. We have analysed regeneration of distinctly different types of intrafusal fibres in rat muscle spindles. We have introduced the new technique of heterochronous allotransplantation and compared it with the previously used standard autografting method. The allotransplantation method enables one to graft muscles from very young animals; we have used the extensor digitorum longus (EDL) muscles from 2- to 28-day-old rats, which were grafted into EDL muscles of adult inbred recipients. The regenerated “intrafusal” fibres did not express the spindle-specific slow tonic and alpha cardiac-like myosin heavy chain (MyHC) isoforms and they did not exhibit the dual mATPase reaction typical of the nuclear bag 2 fibres and the characteristic regional differences in MyHC expression and in the mATPase reaction of nuclear bag 1 and nuclear bag 2 fibres. On the other hand, they expressed either fast twitch or slow twitch/beta cardiac MyHC isoforms and exhibited an alkali or acid stable mATPase reaction along their whole length, like extrafusal fast type 2 and slow type 1 muscle fibres, respectively. In all regenerated muscle spindles only motor, but no sensory axons were found. More than 85% of muscle spindles in our sample contained regenerated spindle fibres of the same extrafusal fibre type (either type 2 or type 1), in contrast to control muscle spindles, which always contained intrafusal fibres of three different intrafusal fibre types (nuclear bag 1, nuclear bag 2 and nuclear chain fibres). There were no differences in MyHC expression and mATPase activity between spindle fibres regenerated in grafts taken from young rats of various ages or between allotransplanted and autotransplanted EDL muscles. The present results demonstrate that regenerated “intrafusal” fibres resemble, according to MyHC expression, extrafusal fast or slow muscle fibres. It can thus be concluded that intrafusal satellite cells derived from distinctly different nuclear bag 1, nuclear bag 2 and nuclear chain fibres show great plasticity, as their MyHC expression can be respecified towards the extrafusal muscle fibre phenotype by foreign alpha-motor innervation.