Kloc Malgorzata, Bilinski Szczepan, Kubiak Jacek Z
The Houston Methodist Research Institute, Houston, TX, USA.
The Houston Methodist Hospital, Department of Surgery, 6550 Fannin St., Houston, TX, 77030, USA.
Methods Mol Biol. 2016;1457:179-90. doi: 10.1007/978-1-4939-3795-0_13.
The majority of oocyte functions involves and depends on the cytoskeletal elements, which include microtubules and actin and cytokeratin filaments. Various structures and molecules are temporarily or permanently bound to the cytoskeletal elements and their functions rely on cytoskeleton integrity and its timely assembly. Thus the accurate visualization of cytoskeleton is often crucial for studies and analyses of oocyte structure and functions. Here we describe several reliable methods for microtubule and/or microfilaments preservation and visualization in Xenopus oocyte extracts, and in situ in live and fixed insect and frog (Xenopus) oocytes. In addition, we describe visualization of cytoskeleton-bound RNAs using molecular beacons in live Xenopus oocytes.
大多数卵母细胞功能涉及并依赖于细胞骨架成分,其中包括微管、肌动蛋白和细胞角蛋白丝。各种结构和分子暂时或永久地与细胞骨架成分结合,其功能依赖于细胞骨架的完整性及其适时组装。因此,准确可视化细胞骨架对于卵母细胞结构和功能的研究与分析往往至关重要。在此,我们描述了几种可靠的方法,用于在非洲爪蟾卵母细胞提取物中以及在活体和固定的昆虫及青蛙(非洲爪蟾)卵母细胞中原位保存和可视化微管和/或微丝。此外,我们还描述了在活体非洲爪蟾卵母细胞中使用分子信标可视化与细胞骨架结合的RNA。