Physiology and Physiopathology of the Gliovascular Unit Research Group, Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS Unité Mixte de Recherche 724, INSERM Unité 1050, Labex Memolife, PSL Research University, Paris, France.
Neuroglial Interactions in Cerebral Physiopathology Research Group, Center for Interdisciplinary Research in Biology (CIRB), College de France, CNRS Unité Mixte de Recherche 724, INSERM Unité 1050, Labex Memolife, PSL Research University, Paris, France.
STAR Protoc. 2020 Dec 4;1(3):100198. doi: 10.1016/j.xpro.2020.100198. eCollection 2020 Dec 18.
Translation of distally localized mRNAs is an evolutionary mechanism occurring in polarized cells. It has been observed in astrocytes, whose processes contact blood vessels and synapses. Here, we describe a protocol for the purification of the entire pool of ribosome-bound mRNAs in perisynaptic astrocytic processes (PAPs). Our procedure combines the preparation of synaptogliosomes with a refined translating ribosome affinity purification technique. This approach can be used in any brain region to probe the physiological relevance of local translation in PAPs. For complete details on the use and execution of this protocol, please refer to Mazaré et al. (2020).
远端定位的 mRNA 翻译是一种发生在极化细胞中的进化机制。这种机制在与血管和突触接触的星形胶质细胞中被观察到。在这里，我们描述了一种在突触旁星形胶质细胞突起（PAPs）中纯化整个核糖体结合的 mRNA 库的方案。我们的程序将突触小体的制备与改良的翻译核糖体亲和纯化技术结合在一起。这种方法可以在任何脑区中用于探测 PAPs 中局部翻译的生理相关性。有关此方案的使用和执行的完整详细信息，请参阅 Mazaré 等人。（2020 年）。
