The purpose of the present study was to determine at which point in the period from embryonic day 21 up to postnatal day (PD) 75, the different fibre types and subtypes are detectable in rat extensor digitorum longus, soleus and gastrocnemius muscles using immunohistochemical, enzyme histochemical and cytophotometrical methods. Moreover, fibre type-specific changes in metabolic profile and changes in fibre type population during postnatal development were analysed. Before birth, no clear differentiation of fibre types was found. At PD 1, slow and fast fibres were typed by antibodies against neonatal, slow and fast myosin heavy chains (MHCs). At PD 8, the different ATPase activities of slow and fast MHCs after alkaline preincubation were detected histochemically. At PD 21, differences in acid stability of ATPase activity of fast MHC isoforms revealed the fast subtypes IIA and IIB (including IIX). At this age, also differences in metabolic properties (oxidative and glycolytic enzyme activities) of fibres were detected for the first time by cytophotometry classifying the fibres into SO, FOG I, FOG II and FG. Before the age of 21 days, the fast fibres were metabolically undifferentiated. During further development and ageing, the population of FG fibres with high glycolytic activity increased at the expense of FOG fibres suggesting FOG to FG transformation. Cytophotometrical measurements revealed that the muscle fibres developed their highest contractile, oxidative and glycolytic activity at PD 21, the time of weaning. In this way, muscle fibres may be prepared for the higher demands for posture and mobility after leaving the nest.