Visualizing Lipophagy as a New Mechanism of the Synthesis of Sex Steroids in Human Ovary and Testis Using Immunofluorescence Staining Method.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2024-02-28 DOI:10.1007/7651_2024_520

Yashar Esmaeilian, Sevgi Yusufoglu, Ece Iltumur, Gamze Bildik, Ozgur Oktem

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Abstract

Immunofluorescence, a transformative tool in cellular biology, is employed to dissect the intricate mechanisms of cholesterol trafficking in human reproductive tissues. Autophagy, a key player in cellular homeostasis, particularly lipophagy, emerges as a free cholesterol source for steroidogenesis. In this chapter, we describe a comprehensive immunofluorescence staining protocol, with details provided for the precise visualization of subcellular dynamics of mitochondria, lysosomes, and lipid droplets in ex vivo testicular tissue and primary luteal granulosa cell culture models, pivotal components in sex steroid biosynthesis. Here, we detail the culture, treatment, and immunofluorescence protocols, providing a comprehensive guide for researchers. The provided immunofluorescence toolkit serves as a valuable resource for researchers, paving way for advancements in human reproductive health to investigate the intricate interplay between autophagy, lipophagy, and cholesterol trafficking.

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用免疫荧光染色法观察嗜脂性是人类卵巢和睾丸合成性激素的新机制

免疫荧光是细胞生物学的一种变革性工具，它被用来剖析人体生殖组织中胆固醇运输的复杂机制。自噬是细胞稳态的关键角色，尤其是噬脂性自噬，是类固醇生成的游离胆固醇来源。在本章中，我们介绍了一种全面的免疫荧光染色方案，详细说明了如何精确观察体外睾丸组织和原代黄体颗粒细胞培养模型中线粒体、溶酶体和脂滴的亚细胞动态，它们是性类固醇生物合成的关键组成部分。在这里，我们详细介绍了培养、处理和免疫荧光方案，为研究人员提供了全面的指南。所提供的免疫荧光工具包是研究人员的宝贵资源，为人类生殖健康研究自噬、噬脂和胆固醇运输之间错综复杂的相互作用铺平了道路。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.