Organotypic slice cultures of dopaminergic neurons of substantia nigra

Abstract

Morphological methods were used to study the plasticity of target-deprived mammalian dopaminergic (DA) neurons. Slices of substantia nigra (SN) were taken from the midbrain of rats aged one to twelve days, and cultured for one to two weeks. Localization of tyrosine hydroxylase (TH) was used to examine the distribution and shapes of DA neurons. Histochemical staining for acetylcholinesterase (AChE) was carried out to estimate both survival and biosynthesis of SN neurons. We found that some DA neurons can survive in vitro without their usual target neurons. This was demonstrated by injecting rhodamine-conjugated microspheres (RD) into the caudate putamen, a SN target area, at 6 to 8 days prior to culturing. RD-labeled cells survived in SN cultures and some of them were doubly labeled with AChE. TH neurons had different shapes and their axon terminals formed close contacts with adjacent nondopaminergic neurons. These findings suggest that a subset of DA neurons may switch targets, but the majority of them require target interactions with the caudate putamen for survival in vitro.