Dissociated cell suspensions were prepared from the substantia nigra of 15–17 day-old rat embryos and grafted via an intraparenchymal injection into the depth of the neostriatum of adult recipient rats. The survival and fibre outgrowth of the dopamine-containing neurones in the implants were studied by fluorescence histochemistry, and the functional capacity of the grafts was monitored by repeated testing of the amphetamine-induced turning behaviour of the implanted rats. Before transplantation the target neostriatum of the recipient rats was denervated of its normal dopaminergic innervation by an injection of 6-hydroxydopamine into the ipsilateral nigrostriatal dopamine pathway. The completeness of the denervation was ascertained by measurement of the intensity of the amphetamine-induced turning response. After injection of the dissociated cells large numbers of dopamine-containing neurones were found in clusters at the site of injection as well as scattered in the apparently intact neostriatal tissue up to a distance of about 0.5 mm from the site of injection. Extensive dopamine-containing fibre networks had developed around the implant. These newly formed fibres, which were most abundant around the cell clusters at the injection site, extended in a loose network into large areas of the initially denervated caudate-putamen. In all animals with surviving dopamine neurones the amphetamine-induced turning response was reduced, and in the most extensively reinnervated cases even reversed, within 3–5 weeks after transplantation. This strongly suggests that the implanted dopamine neurones are capable of restoring dopaminergic neurotransmission in the denervated neostriatum, probably via reinnervation of the denervated neostriatal tissue. The use of dissociated brain tissue preparations thus permits reliable intraparenchymal grafting of neurones to plausibly any desired site within the central nervous system, and should open entirely new possibilities for investigation of neuronal growth dynamics and functional reconstruction of damaged brain circuits, perhaps even in brains of larger mammals.
Cell and Tissue Research – Springer Journals
Published: Nov 1, 1980
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