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

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....

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.)
Main Authors Esmaeilian, Yashar, Yusufoglu, Sevgi, Iltumur, Ece, Bildik, Gamze, Oktem, Ozgur
Format Journal Article
LanguageEnglish
Published United States 28.02.2024
Subjects
ImageJ
Immunofluorescence staining
Luteinized granulosa cells
Testis
Confocal microscopy
Steroidogenesis
Autophagy
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Summary: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.
ISSN:1940-6029
DOI:10.1007/7651_2024_520