Protocol for the purification and transcriptomic analysis of mouse astrocytes using GFAT.
Protocol for the purification and transcriptomic analysis of mouse astrocytes using GFAT.
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使用GFAT纯化和转录组学分析小鼠星形胶质细胞的方案。
DOI：
10.1016/j.xpro.2023.102599
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发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Allen, Nicola J.
中科院分区：
文献类型：
--
作者：
Labarta-Bajo, Lara;Deng, James;Contreras, Minerva;Allen, Nicola J.
星形胶质细胞是中枢神经系统中调节神经元功能的神经胶质细胞。在这里，我们提出了糖基化固定的星形胶质细胞核转录组学（GFAT），从成年和老年新鲜小鼠大脑的皮质和小脑的星形胶质细胞核的纯化和转录组学分析的协议。我们描述了组织解剖、乙二醛固定、均质化、细胞核分离、抗体染色、荧光激活细胞分选和RT-qPCR或批量RNA测序的步骤。GFAT不需要转基因系或病毒注射，并允许平行的星形胶质细胞和神经元分析。使用RT-qPCR和RNA-seq纯化和分析星形胶质细胞核小鼠模型、脑区和年龄中的星形胶质细胞转录组学平行神经元分析，可以发现神经元-星形胶质细胞串扰出版商说明：进行任何实验方案都需要遵守当地实验室安全和道德机构指南。星形胶质细胞是中枢神经系统中调节神经元功能的神经胶质细胞。在这里，我们提出了糖基化固定的星形胶质细胞核转录组学（GFAT），从成年和老年新鲜小鼠大脑的皮质和小脑的星形胶质细胞核的纯化和转录组学分析的协议。我们描述了组织解剖、乙二醛固定、均质化、细胞核分离、抗体染色、荧光激活细胞分选和RT-qPCR或批量RNA测序的步骤。GFAT不需要转基因系或病毒注射，并允许平行的星形胶质细胞和神经元分析。
Astrocytes are glial cells of the central nervous system that modulate neuronal function. Here, we present glyoxal-fixed astrocyte nuclei transcriptomics (GFAT), a protocol for the purification and transcriptomic analysis of astrocyte nuclei from the cortex and cerebellum of adult and aged fresh mouse brain. We describe steps for tissue dissection, glyoxal fixation, homogenization, nuclei isolation, antibody staining, fluorescence-activated cell sorting, and RT-qPCR or bulk RNA sequencing. GFAT does not require transgenic lines or viral injection and allows parallel astrocyte and neuron profiling. Purification and analysis of astrocyte nuclei with RT-qPCR and RNA-seq Astrocyte transcriptomics across mouse models, brain regions, and ages Parallel neuron profiling, which can uncover neuron-astrocyte crosstalk Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Astrocytes are glial cells of the central nervous system that modulate neuronal function. Here, we present glyoxal-fixed astrocyte nuclei transcriptomics (GFAT), a protocol for the purification and transcriptomic analysis of astrocyte nuclei from the cortex and cerebellum of adult and aged fresh mouse brain. We describe steps for tissue dissection, glyoxal fixation, homogenization, nuclei isolation, antibody staining, fluorescence-activated cell sorting, and RT-qPCR or bulk RNA sequencing. GFAT does not require transgenic lines or viral injection and allows parallel astrocyte and neuron profiling.
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影响因子：
16.6
作者：
Batiuk, Mykhailo Y.;Martirosyan, Araks;Holt, Matthew G.
通讯作者：
Holt, Matthew G.
来自小鼠大脑多个区域的老化星形胶质细胞转录组。
影响因子：
8.8
作者：
Boisvert MM;Erikson GA;Shokhirev MN;Allen NJ
通讯作者：
Allen NJ
影响因子：
7.7
作者：
Farhy-Tselnicker I;Boisvert MM;Liu H;Dowling C;Erikson GA;Blanco-Suarez E;Farhy C;Shokhirev MN;Ecker JR;Allen NJ
通讯作者：
Allen NJ
影响因子：
16
作者：
Heinz S;Benner C;Spann N;Bertolino E;Lin YC;Laslo P;Cheng JX;Murre C;Singh H;Glass CK
通讯作者：
Glass CK
影响因子：
13.6
作者：
Herrero-Navarro Á;Puche-Aroca L;Moreno-Juan V;Sempere-Ferràndez A;Espinosa A;Susín R;Torres-Masjoan L;Leyva-Díaz E;Karow M;Figueres-Oñate M;López-Mascaraque L;López-Atalaya JP;Berninger B;López-Bendito G
通讯作者：
López-Bendito G