González-Jamett Arlek, Maldifassi María Constanza, Cárdenas Ana María
Centro Interdisciplinario de Neurociencia de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso, Chile.
Escuela de Química y Farmacia, Facultad de Farmacia, Universidad de Valparaíso, Valparaíso, Chile.
Methods Mol Biol. 2023;2565:283-296. doi: 10.1007/978-1-0716-2671-9_19.
Chromaffin granules isolated from adrenal glands constitute a powerful experimental tool to the study of secretory vesicle components and their participation in fusion and docking processes, vesicle aggregation, and interactions with cytosolic components. Although it is possible to isolate and purify chromaffin granules from adrenal glands of different species, bovine adrenal glands are the most used tissue source due to its easy handling and the large amount of granules that can be obtained from this tissue. In this chapter, we describe an easy-to-use and short-term protocol for efficiently obtaining highly purified chromaffin granules from bovine adrenal medulla. We additionally include protocols to isolate granules from cultured bovine chromaffin cells and PC12 cells, as well as a section to obtain chromaffin granules from mouse adrenal glands.
从肾上腺分离出的嗜铬颗粒是研究分泌囊泡成分及其在融合和对接过程、囊泡聚集以及与胞质成分相互作用中所起作用的强大实验工具。虽然可以从不同物种的肾上腺中分离和纯化嗜铬颗粒,但由于牛肾上腺易于操作且能从该组织中获得大量颗粒,因此它是最常用的组织来源。在本章中,我们描述了一种易于使用的短期方案,用于从牛肾上腺髓质高效获得高度纯化的嗜铬颗粒。我们还包括从培养的牛嗜铬细胞和PC12细胞中分离颗粒的方案,以及从小鼠肾上腺中获取嗜铬颗粒的章节。