Target specificity of dopaminergic neurons in dissociated cultures from the Substantianigra and the ventral tegmental area

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1.Separate cell cultures of the striatum (ST) and the nucleus accumbens (AC) : After making brain slices, the regions of the ST and the AC were separately dissected from three-day-old postnatal rats. The fragments of brain tissue were dissociated and cultured on a glia feeder layr. Immunohistochemical staining of these cultures revealed that many of cultured neurons were GABAergic. Morphological characteristics of the GABA neurons in the ST culture and in the AC culture were different each other, and were also different from that of the substantia nigra (SN) culture.2.Tissue specificity of glial effects on the separate cell cultures : Various combination cultures of the separated neurons and glial cells from different brain regions revealed that the SN neurons as well as the ST neurons grew well on glial cell layr prepared from the brain tissue of their own regions or target regions but not on glial cells from non-target areas.3.Synapse formation between GABA ergic neurons and dopamine (DA) ergic neurons in the separate culture : Electron microscopy using immunohistochemistry for GABA or tyrosine hydroxylase (TH) revealed that synaptic contacts were made reciprocally between GABA ergic and DA ergic neurons in the SN culture. Further study should be made to quantify the synapse formation in various combinations of co-culture, such as in SN-ST culture.4.Correlation of DA ergic terminals and nitric oxide (NO) neurons in the rat striatum. Electron microscopic study combined with TH immunohistochemistry and NADPH-diaphorase histochemistry demonstrated an association of DA ergic terminal and possible glutamatergic terminals which was directry apposing to the NO neurons, suggesting the site of NMDA receptor-evoked NO release which in turn induce DA release from the DA terminal.