Herp depletion arrests the S phase of the cell cycle and increases estradiol synthesis in mouse granulosa cells

The Journal of Reproduction and Development
Fenglei ChenYaPing Jin

Abstract

The endoplasmic reticulum (ER) stress response has been implicated in the development, atresia and luteinization of ovarian follicles. However, there have been few reports concerning the role of Herp, an ER stress-induced protein, in follicular development. The present study aims to detect the distribution and cyclic variations of Herp during the estrous cycle and to reveal the roles of Herp in regulating the cell cycle, apoptosis and steroid hormone biosynthesis in mouse granulosa cells. In this study, immunohistochemistry staining showed that Herp expression was primarily in the granulosa cells and oocytes. Furthermore, we constructed recombinant lentiviral vectors for Herp short hairpin interfering RNA (shRNA) expression; immunofluorescence staining, real-time quantitative PCR (RT-qPCR) and western blot analysis revealed that Herp was successfully knocked down. Flow cytometry showed that knockdown of Herp arrested granulosa cells at the S phase of the cell cycle. More importantly, ELISA analysis revealed that Herp knockdown significantly upregulated the concentration of estradiol (E2) in the culture supernatants. RT-qPCR was performed to determine the regulatory mechanism of Herp knockdown in the cell cycle, and in steroid s...Continue Reading

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Citations

Jul 13, 2018·The Journal of Reproduction and Development·Lanjie LeiLei Yang
Feb 6, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yongjie XiongShaojun He
May 8, 2020·Molecular and Cellular Endocrinology·Jiahui HuFenglei Chen
Nov 17, 2021·Journal of Zhejiang University. Science. B·Zhinan ZhouLei Hong

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Methods Mentioned

BETA
protein folding
PCR
Protein Assay
ELISA
Fluorescence
flow cytometry

Software Mentioned

Quantity One
SPSS ( Statistical Package for the Social Sciences )
SPSS

Related Concepts

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Apoptosis

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