Development of the cholinergic fibres innervating the cerebral cortex of the rat

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Abstract
The ontogeny of innervation of the cholinergic fibres from the basal forebrain into the cingulate, frontal, parietal and piriform cortices of the rat has been examined using a modified histochemical method of acetylcholinesterase (AChE). The method produced crisp fibre staining with enhanced visibility and a clear back-ground, and a pattern of the distribution of these fibres was comparable to that achieved by choline acetyltransferase (ChAT) immunocytochemistry. In the rat, the AChE-stained fibres developed progressively from the deep cortical white matter towards the cortex itself. In general, a few AChE-positive fibres were seen in the subcortical white matter and the cingulum bundle, entering into the cerebral cortex by about 5 postnatal days. The number of these AChE-positive processes increased dramatically during the following two weeks. Thereafter, the general appearance of the overall pattern of distribution of the AChE fibres changed little, but the staining density became gradually more intense and by about 28 days after birth it was virtually indistinguishable from that in the adult. The onset and the development of the AChE-positive fibre network varied considerably between individual cortical regions, and indicated, in general, an anterior to posterior gradient. Within the dispersed AChE fibre network in the cerebral cortex, three bands of relatively enriched cholinergic processes, namely the deep cortical, mid-cortical and superficial layers, developed in an 'inside-out' fashion. The exact position of some of these AChE-rich bands varied from one cortical region to another and during development. A striking correlation during ontogeny was observed in the cerebral cortex between the changing patterns of AChE fibre network and the activity of ChAT, the enzyme synthesizing acetylcholine. The present findings can also provide an important anatomical baseline for future studies related to the factors controlling the expression of ChAT activity and the development of cholinergic neurotransmitter system in the rat.