Real-Time Visualization of Cholesterol Trafficking in Human Granulosa Cells Using Confocal Live Cell Microscopy as a Tool to Study the Novel Role of Autophagy in Sex Steroid Synthesis.

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2024-02-28 DOI:10.1007/7651_2024_521
Yashar Esmaeilian, Ozgur Oktem
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Abstract

Autophagy is an evolutionarily conserved process that aims to maintain the energy homeostasis of the cell by recycling long-lived proteins and organelles. We have very recently demonstrated that lipophagy, a special form of autophagy, mediates the association of the lipid droplets (LDs) with lysosomes to deliver the lipid cargo within the LDs to lysosomes for degradation in order to release free cholesterol required for steroid synthesis in human ovary and testis. In this chapter, we describe live cell confocal microscopy technique that allows us to monitor real-time cholesterol trafficking and the association of cholesterol-laden LDs with lysosome (lipophagy) in human granulosa cells.

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利用共聚焦活细胞显微镜实时观察人类颗粒细胞中胆固醇的迁移,研究自噬在性类固醇合成中的新作用。
自噬是一种进化保守过程,旨在通过回收长寿命蛋白质和细胞器来维持细胞的能量平衡。我们最近证明,自噬的一种特殊形式--噬脂作用介导脂滴(LDs)与溶酶体的结合,将脂滴内的脂质货物运送到溶酶体降解,以释放人类卵巢和睾丸合成类固醇所需的游离胆固醇。在本章中,我们将介绍活细胞共聚焦显微镜技术,该技术使我们能够实时监测人粒细胞中胆固醇的运输以及胆固醇载脂小体与溶酶体的结合(噬脂作用)。
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来源期刊
Methods in molecular biology
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.
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