Induction of axon and dendrite formation during early RGC-5 cell differentiation
Induction of axon and dendrite formation during early RGC-5 cell differentiation
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DOI：
10.1016/j.exer.2007.08.001
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发表时间：
2007-11-01
影响因子：
3.4
通讯作者：
Levin, Leonard A.
中科院分区：
文献类型：
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作者：
Lieven, Christopher J.;Millet, Lucia E.;Levin, Leonard A.
星形孢菌素可分化视网膜神经节细胞 (RGC) 样 RGC-5 系，以停止分裂、延长神经突并增加多种神经节细胞标记物的水平。这使得可以在单细胞的基础上研究神经突发育的调节。然而，目前尚不清楚分化诱导的神经突是否具有树突或轴突的特征。为了解决这个问题，用十字孢菌素分化 RGC-5 细胞，然后对微管相关蛋白 2 (MAP2) 和肌动蛋白进行免疫印迹，或对不同的 MAP2 亚型、tau、生长相关蛋白 43 (GAP-43) 进行免疫细胞化学染色。或神经元标记物β-III-微管蛋白。我们发现星形孢菌素诱导的分化导致 MAP2c 的上调，MAP2c 是发育中神经元中表达的 MAP2 亚型。一些神经突表达 MAP2c，但不表达树突标记物 MAP2a 和 MAP2b，这与轴突表型一致。一些神经突以特征性的近端到远端梯度表达轴突标记 tau，并具有轴突生长锥的 GAP-43 标记特征。分化的 RGC-5 细胞中 MAP2c 的存在表明 RGC 样神经突发育，不同 MAP2 同工型的染色模式以及 tau 和 GAP-43 的阳性表明分化诱导轴突样和树突样神经突。 (C) 2007 Elsevier Ltd. 保留所有权利。
The retinal ganglion cell (RGC)-like RGC-5 line can be differentiated with staurosporine to stop dividing, extend neurites, and increase levels of several ganglion cell markers. This allows study of regulation of neurite development on a single cell basis. However, it is unclear whether the neurites induced by differentiation have features characteristic of dendrites or axons. To address this question, RGC-5 cells were differentiated with staurosporine and then immunoblotted for microtubule-associated protein 2 (MAP2) and actin, or stained immunocytochemically for different MAP2 isoforms, tau, growth-associated protein 43 (GAP-43). or the neuronal marker beta-III-tubulin. We found that staurosporine-induced differentiation led to an upregulation of MAP2c, a MAP2 isoform expressed in developing neurons. Some neurites expressed MAP2c but not the dendritic markers MAP2a and MAP2b, consistent with an axonal phenotype. Some neurites expressed the axonal marker tau in a characteristic proximal-to-distal gradient, and had GAP-43 labeling characteristic of axonal growth cones. The presence of MAP2c in differentiated RGC-5 cells is indicative of RGC-like neurite development, and the pattern of staining for the different MAP2 isoforms, as well as positivity for tau and GAP-43, indicates that differentiation induces axon-like and dendrite-like neurites. (C) 2007 Elsevier Ltd. All rights reserved.