Alejandre-García Tzitzitlini, Segura-Chama Pedro, Parada-Parra Oscar Javier, Millán-Aldaco Diana, Hernández-Cruz Arturo
Department of Biological Sciences, Columbia University, New York, NY, USA.
Cátedras CONACyT, Laboratorio de Fisiología Celular, Instituto Nacional de Psiquiatría Ramón de La Fuente Muniz, México City, Mexico.
Methods Mol Biol. 2023;2565:129-151. doi: 10.1007/978-1-0716-2671-9_10.
The spontaneously hypertensive rat (SHR) is a model widely used to investigate the causal mechanisms of essential hypertension. The enhanced catecholamine (CA) release reported in adrenal glands from adult SHRs raised considerable interest for its possible implication in the genesis of hypertension. The use of powerful techniques such as calcium imaging, electrophysiology, and single-cell amperometry to monitor in real time the key steps in CA secretion has allowed a better understanding of the role of chromaffin cells (CC) in the pathophysiology of hypertension, although several questions remain. Additionally, the implementation of these techniques in preparations in situ, such as the acute adrenal gland slice, which maintains the microenvironment, cell-to-cell communication, and anatomical structure similar to that of the intact adrenal gland, yields data that may have even greater physiological relevance. Here, we describe the procedures to measure the blood pressure of rats in a noninvasive manner, how to obtain primary cultures of adrenal chromaffin cells and acute adrenal slices, and how to perform amperometric recordings and intracellular calcium imaging in these preparations.
自发性高血压大鼠（SHR）是一种广泛用于研究原发性高血压病因机制的模型。成年SHR肾上腺中报道的儿茶酚胺（CA）释放增强因其可能与高血压的发生有关而引起了极大关注。使用钙成像、电生理学和单细胞安培法等强大技术实时监测CA分泌的关键步骤，有助于更好地理解嗜铬细胞（CC）在高血压病理生理学中的作用，尽管仍有几个问题存在。此外，在诸如急性肾上腺切片等原位制剂中实施这些技术，该制剂保持了与完整肾上腺相似的微环境、细胞间通讯和解剖结构，所产生的数据可能具有更大的生理相关性。在这里，我们描述了以非侵入性方式测量大鼠血压的程序、如何获得肾上腺嗜铬细胞的原代培养物和急性肾上腺切片，以及如何在这些制剂中进行安培记录和细胞内钙成像。
