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胶质细胞对大鼠腹侧中脑器官型组织培养中神经纤维形成的影响。

Glial influence on nerve fiber formation from rat ventral mesencephalic organotypic tissue cultures.

作者信息

Berglöf Elisabet, Af Bjerkén Sara, Strömberg Ingrid

机构信息

Department of Integrative Medical Biology, Umeå University, Umeå, Sweden.

出版信息

J Comp Neurol. 2007 Mar 20;501(3):431-42. doi: 10.1002/cne.21251.

Abstract

Rat fetal ventral mesencephalic organotypic cultures have demonstrated two morphologically different dopamine nerve fiber growth patterns, in which the initial nerve fibers are formed in the absence of astrocytes and the second wave is guided by astrocytes. In this study, the presence of subpopulations of dopamine neurons, other neuronal populations, and glial cells was determined. We used "roller-drum" organotypic cultures, and the results revealed that beta-tubulin-positive/tyrosine hydroxylase (TH)-negative nerve fibers were present as early as 1 day in vitro (DIV). A similar growth pattern produced by TH-positive neurons was present from 2 DIV. These neurites grew to reach distances over 4 mm and over time appeared to be degenerating. Thin, vimentin-positive processes were found among these nerve fibers. As the first growth was retracted, a second outgrowth was initiated and formed on migrating astrocytes. TH- and aldehyde dehydrogenase-1 (ALDH1)-positive nerve fibers formed both the nonglia-associated and the glia-associated outgrowth. In cultures with membrane inserts, only the glia-associated outgrowth was found. Vimentin-positive cells preceded migration of NG2-positive oligodendrocytes and Iba-1-positive microglia. Oligodendrocytes appeared not to be involved in guiding neuritic growth, but microglia was absent over areas dense with TH-positive neurons. In conclusion, in "roller-drum" cultures, nerve fibers are generally formed in two sequences. The early-formed nerve fibers grow in the presence of thin, vimentin-positive processes. The second nerve fiber outgrowth is formed on astroglia, with no correlation to the presence of oligodendrocytes or microglia. ALDH1-positive nerve fibers, presumably derived from A9 dopamine neurons, participate in formation of both sequences of outgrowth.

摘要

大鼠胚胎腹侧中脑器官型培养物已显示出两种形态不同的多巴胺神经纤维生长模式,其中初始神经纤维在无星形胶质细胞的情况下形成,第二波由星形胶质细胞引导。在本研究中,确定了多巴胺神经元亚群、其他神经元群体和神经胶质细胞的存在。我们使用了“滚筒”器官型培养物,结果显示,早在体外培养1天(DIV)时就存在β-微管蛋白阳性/酪氨酸羟化酶(TH)阴性神经纤维。从培养2天开始出现由TH阳性神经元产生的类似生长模式。这些神经突生长至超过4毫米的距离,随着时间的推移似乎在退化。在这些神经纤维中发现了纤细的波形蛋白阳性突起。随着第一次生长的回缩,第二次生长开始并在迁移的星形胶质细胞上形成。TH和醛脱氢酶-1(ALDH1)阳性神经纤维形成了与神经胶质细胞无关和与神经胶质细胞相关的生长。在有膜插入物的培养物中,仅发现了与神经胶质细胞相关的生长。波形蛋白阳性细胞先于NG2阳性少突胶质细胞和Iba-1阳性小胶质细胞的迁移。少突胶质细胞似乎不参与引导神经突生长,但在TH阳性神经元密集的区域没有小胶质细胞。总之,在“滚筒”培养物中,神经纤维通常按两个顺序形成。早期形成的神经纤维在纤细的波形蛋白阳性突起存在的情况下生长。第二次神经纤维生长在星形胶质细胞上形成,与少突胶质细胞或小胶质细胞的存在无关。ALDH1阳性神经纤维,可能源自A9多巴胺神经元,参与了两个生长顺序的形成。

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