Elimination of distributed acetylcholine receptor clusters from developing fast-twitch fibres in an avian muscle

The development of the focal localization of large acetylcholine receptor clusters (AChR-Cs) on avian fast muscle fibres has been investigated in the triceps brachii pars humeralis (TH) muscle of the chick embryo. The mature TH muscle consists of both fast fibres, which usually receive a focal innervation at single synaptic sites, and slow fibres which receive a distributed innervation at multiple synaptic sites. Single fibre fragments dissociated from the embryonic muscle were typed using anti-myosin antibodies; fluorescently labelled alpha-bungarotoxin was used to identify large AChR-Cs which serve as synaptic markers. In contrast to the mature focal innervation, at embryonic day 11 (E11), many fast-type fibres in the TH muscle displayed large, distributed AChR-Cs (3.7 +/- 0.7 per 1000 microns fibre length; n = 6 embryos) like neighbouring slow-type fibres. By E16 distributed AChR-Cs were rare on fast type fibres (0.9 +/- 0.2 per 1000 microns fibre length). As it was possible that the frequency of fast fibres with distributed AChR-Cs declined simply as a consequence of the increase in number of secondary generation fibres, tritiated thymidine was injected at E7 in order to identify the primary generation fibres at E14. The great majority of fast fibres that were heavily labelled with thymidine at E14 appeared to possess a focal AChR-C. The results suggest that at E11 fast-type primary fibres in the TH muscle receive a distributed innervation very similar to neighbouring slow-type fibres; this subsequently evolves into the mature focal innervation following the elimination of synaptic sites between E11 and E14.