Regional differences in organization of the extracellular matrix and cytoskeleton at the equator of chicken intrafusal muscle fibres.

Abstract

Equatorial regions of chicken intrafusal fibres were examined with a panel of monoclonal antibodies against intracellular proteins and components of extracellular matrix to identify structural associations at points of contact between sensory terminals and intrafusal fibres, and at points which lacked them. One aspect of this study was to establish whether the known morphological differences between myosensory and neuromuscular junctions also extended to the molecular level. As viewed in cross-sections, myosensory junctions at the equator are restricted to approximately one-half of the intrafusal fibre circumference, a region referred to as the sensory sector. The diametrically opposite region which lacks sensory terminals is referred to as the non-sensory sector. The basal lamina over the sensory sector was positive for chondroitin sulphate, while that part which covered the non-sensory sector was negative. Staining for collagen type IV was very faint at the sensory sector and stronger at the non-sensory sector, but immunoreactivity for heparan sulphate proteoglycan and laminin was moderate to strong in all parts of the basal lamina. Within intrafusal fibres, filamin and alpha-actinin were largely limited to the sensory sector. The major feature of the non-sensory sector was a sharply delineated, narrow intrafibre crescent of vinculin, and colocalized with it, a crescent of talin. The plasmalemma of intrafusal fibres at the non-sensory sector reacted positively for the beta 1 subunit of the integrin family of receptors. Immunolocalization of these receptors was not observed to any significant extent in the sensory sector. Towards the end of the equator and the initial portion of the juxtaequator, chondroitin sulphate, vinculin and the other proteins came gradually to be distributed equally all the way round the intrafusal fibres. This changeover in distribution of connective tissue proteins and structural intracellular proteins parallels the decreasing number of contacts made by sensory terminals.