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

黑质和腹侧被盖区分离培养物中多巴胺能神经元的靶特异性

基本信息

批准号：
05680662
负责人：
MASUKO Sadahiko
金额：
$ 1.28万
依托单位：
Saga Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

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.

1.纹状体（ST）和中脑核（AC）的单独细胞培养：制作脑切片后，从出生后3天的大鼠中分别解剖ST和AC区域。将脑组织碎片分离并培养在胶质细胞饲养层上。免疫组织化学染色显示，许多培养的神经元是GABA能的。ST培养和AC培养中GABA神经元的形态特征相互不同，也与黑质（SN）培养中的GABA神经元不同。2.胶质细胞对单独细胞培养物影响的组织特异性：不同脑区分离的神经元与胶质细胞的各种组合培养显示，SN神经元和ST神经元在胶质细胞上生长良好3.分离培养的GABA能神经元与多巴胺（DA）能神经元之间的突触形成：电镜下GABA或酪氨酸羟化酶（TH）免疫组化显示，在SN培养中，GABA能神经元与DA能神经元之间形成突触接触。进一步的研究还需要对各种联合培养条件下的突触形成进行定量研究，如SN-ST培养。4.大鼠纹状体DA能神经末梢与一氧化氮（NO）神经元的相关性研究。电镜结合TH免疫组化和NADPH黄递酶组织化学研究表明，DA能终末和可能的NMDA能终末与NO神经元直接对置，提示NMDA受体诱发NO释放的部位，进而诱导DA能终末释放DA。