Muscle biopsies were taken at the oesophageal opening and in the costal part of the diaphragm from 15 foetuses (aged between 11 and 40 weeks) and from four children aged 1 week, 5 months, 3 and 16 years. Subjects with multiple congenital abnormalities or neuromuscular diseases were not included in this study. Enzyme histochemistry (myosin ATPases) and immunohistochemistry using antibodies directed against vimentin, desmin, titin and different isoforms of the myosin heavy chains (MHCs) were carried out on serial frozen sections. The diameters of the different types of fibres were measured using a Leitz ASM 68K and the corresponding results were expressed either as mean values or as histograms. In both of the regions of the diaphragm which were examined (oesophageal opening and costal portion). The skeletal muscle fibres are formed from two successive generations in a manner very similar to that which we have previously described for the limb muscles. The chronology of the various events involved in the maturation of the diaphragm does however present certain distinctive features since the adult MHCs are expressed well before 30 weeks even though paradoxically large amounts of fetal MHC were still present at birth. These findings suggest the existence of a special regulatory mechanism in the diaphragm resulting either from its innervation by the phrenic nerve or from its specific functional constraints.
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