Localization of mRNA in vertebrate axonal compartments by in situ hybridization.
Localization of mRNA in vertebrate axonal compartments by in situ hybridization.
复制标题
通过原位杂交在脊椎动物轴突区室中定位 mRNA。
DOI：
10.1007/978-1-61779-005-8_8
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Sotelo,JoseRoberto
中科院分区：
文献类型：
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作者：
Sotelo-Silveira,JoseRoberto;Calliari,Aldo;Kun,Alejandra;Elizondo,Victoria;Canclini,Lucia;Sotelo,JoseRoberto
RNA在脊椎动物轴突中的原位杂交（ISH）的结论性证明一直难以捉摸。我们回顾困难的最重要的原因，包括轴突RNA的浓度低，在特定的皮质域的本地化，并需要隔离轴突。我们证明了轴突显微解剖的重要性，以获得一个整体的安装角度的mRNA分布在轴突领土。我们描述了一种在分离的轴突中进行荧光ISH的方案和用于保存皮质含RNA结构域（例如，轴浆周核糖体斑块或PARP）。
The conclusive demonstration of RNA in vertebrate axons by in situ hybridization (ISH) has been elusive. We review the most important reasons for difficulties, including low concentration of axonal RNAs, localization in specific cortical domains, and the need to isolate axons. We demonstrate the importance of axon micro-dissection to obtain a whole mount perspective of mRNA distribution in the axonal territory. We describe a protocol to perform fluorescent ISH in isolated axons and guidelines for the preservation of structural and molecular integrity of cortical RNA-containing domains (e.g., Periaxoplasmic Ribosomal Plaques, or PARPs) in isolated axoplasm.