Brain Research Institute, University of Zurich, Zurich 8057, Switzerland.
Department of Genetics and Development, Columbia University Medical Center, New York, NY 10033, USA.
STAR Protoc. 2021 Jul 27;2(3):100695. doi: 10.1016/j.xpro.2021.100695. eCollection 2021 Sep 17.
This protocol describes the isolation and culturing of primary neural stem cells (NSCs) from the adult mouse hippocampus, followed by the experimental approach for fluorescence loss in photobleaching assays, previously used to characterize the presence of an endoplasmic reticulum (ER) membrane diffusion barrier. The assay described here can be used to study live asymmetry in the ER membrane or other organelles that is established in dividing NSCs. For complete details on the use and execution of this protocol, please refer to Clay et al. (2014); bin Imtiaz et al. (2021); Lee et al. (2016); Luedeke et al. (2005); Moore et al. (2015); Shcheprova et al. (2008).
本方案描述了从成年小鼠海马体中分离和培养原代神经干细胞（NSCs），随后进行荧光漂白实验的实验方法，该方法先前用于表征内质网（ER）膜扩散屏障的存在。这里描述的测定可用于研究在分裂的 NSCs 中建立的 ER 膜或其他细胞器的活不对称性。有关该方案使用和执行的完整详细信息，请参考 Clay 等人（2014 年）；bin Imtiaz 等人（2021 年）；Lee 等人（2016 年）；Luedeke 等人（2005 年）；Moore 等人（2015 年）；Shcheprova 等人（2008 年）。
