Development of cerebellar neurons and glias revealed by in utero electroporation: Golgi-like labeling of cerebellar neurons and glias.
Development of cerebellar neurons and glias revealed by in utero electroporation: Golgi-like labeling of cerebellar neurons and glias.
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DOI：
10.1371/journal.pone.0070091
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Murakami F
中科院分区：
文献类型：
--
作者：
Kita Y;Kawakami K;Takahashi Y;Murakami F
小脑皮层的功能依赖于由几种不同类型的神经元和胶质细胞组成的精确排列的细胞结构。研究表明，小脑兴奋性神经元和抑制性神经元具有不同的空间起源，分别位于上菱形唇区（uRL）和心室区（VZ），并且不同类型的神经元具有不同的诞生日期。然而，由于技术限制，uRL/VZ祖细胞与其最终表型之间的时空关系仍然知之甚少。为了解决这个问题，我们使用小鼠胚胎在子宫内电穿孔（IUE）荧光蛋白质粒标记特定发育阶段的uRL/VZ祖细胞，并观察成熟时标记的细胞。为了克服由祖细胞分裂引起的质粒的任何潜在稀释，我们还使用了能够永久标记细胞的结构。小脑神经元和胶质细胞以类似高尔基的方式进行标记，使标记细胞能够立即识别。用常规质粒标记5种类型的小脑神经元，即浦肯野、高尔基、卢加罗和单极刷状细胞、大直径深核（DN）神经元和DN星形胶质细胞，而能够永久标记的质粒还可以标记星状细胞、篮状细胞和颗粒细胞以及3种胶质细胞。IUE允许我们标记不同发育阶段的uRL/VZ祖细胞。我们发现5种类型的神经元和DN星形胶质细胞的标记与IUE分期相关，而星形细胞、篮状细胞、颗粒细胞和3种胶质细胞的标记与IUE分期无关。因此，IUE是一种有效的方法来跟踪uRL/VZ祖细胞朝向脑室管腔的小脑细胞的发育。他们还表明，虽然uRL/VZ祖细胞的五种类型神经元的产生以一种时间依赖性的方式受到调节，但祖细胞池在整个胚胎发育过程中保持多能性。
Cerebellar cortical functions rely on precisely arranged cytoarchitectures composed of several distinct types of neurons and glias. Studies have indicated that cerebellar excitatory and inhibitory neurons have distinct spatial origins, the upper rhombic lip (uRL) and ventricular zone (VZ), respectively, and that different types of neurons have different birthdates. However, the spatiotemporal relationship between uRL/VZ progenitors and their final phenotype remains poorly understood due to technical limitations. To address this issue, we performed in utero electroporation (IUE) of fluorescent protein plasmids using mouse embryos to label uRL/VZ progenitors at specific developmental stages, and observed labeled cells at maturity. To overcome any potential dilution of the plasmids caused by progenitor division, we also utilized constructs that enable permanent labeling of cells. Cerebellar neurons and glias were labeled in a Golgi-like manner enabling ready identification of labeled cells. Five types of cerebellar neurons, namely Purkinje, Golgi, Lugaro and unipolar brush cells, large-diameter deep nuclei (DN) neurons, and DN astrocytes were labeled by conventional plasmids, whereas plasmids that enable permanent labeling additionally labeled stellate, basket, and granule cells as well as three types of glias. IUE allows us to label uRL/VZ progenitors at different developmental stages. We found that the five types of neurons and DN astrocytes were labeled in an IUE stage-dependent manner, while stellate, basket, granule cells and three types of glias were labeled regardless of the IUE stage. Thus, the results indicate the IUE is an efficient method to track the development of cerebellar cells from uRL/VZ progenitors facing the ventricular lumen. They also indicate that while the generation of the five types of neurons by uRL/VZ progenitors is regulated in a time-dependent manner, the progenitor pool retains multipotency throughout embryonic development.
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DOI：
10.1016/j.mcn.2011.11.008
发表时间：
2012-02
期刊：
Molecular and cellular neurosciences
影响因子：
--
作者：
Hagan N;Zervas M
通讯作者：
Zervas M
影响因子：
2.5
作者：
ALTMAN, J;BAYER, SA
通讯作者：
BAYER, SA
DOI：
10.1073/pnas.0910786107
发表时间：
2010-06-08
影响因子：
11.1
作者：
Chizhikov, Victor V.;Lindgren, Anne G.;Millen, Kathleen J.
通讯作者：
Millen, Kathleen J.
影响因子：
2.5
作者：
BATINI, C;COMPOINT, C;GUEGAN, M
通讯作者：
GUEGAN, M
影响因子：
2.7
作者：
Grimaldi, Piercesare;Parras, Carlos;Wassef, Marion
通讯作者：
Wassef, Marion