An immortalized steroidogenic goat granulosa cell line as a model system to study the effect of the endoplasmic reticulum (ER)-stress response on steroidogenesis

Diqi YANG; Lei WANG; Pengfei LIN; Tingting JIANG; Nan WANG; Fan ZHAO; Huatao CHEN; Keqiong TANG; Dong ZHOU; Aihua WANG; Yaping JIN

doi:10.1262/jrd.2016-111

Original Article

An immortalized steroidogenic goat granulosa cell line as a model system to study the effect of the endoplasmic reticulum (ER)-stress response on steroidogenesis

Diqi YANG, Lei WANG, Pengfei LIN, Tingting JIANG, Nan WANG, Fan ZHAO, Huatao CHEN, Keqiong TANG, Dong ZHOU, Aihua WANG, Yaping JIN

Author information

Diqi YANG
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Lei WANG
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Pengfei LIN
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Tingting JIANG
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Nan WANG
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Fan ZHAO
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Huatao CHEN
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Keqiong TANG
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Dong ZHOU
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Aihua WANG
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China
Yaping JIN
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, 712100, Shaanxi, China
College of Veterinary Medicine, Northwest A&F University, 712100, Shaanxi, China

Keywords: Endoplasmic reticulum stress (ERS), Granulosa cells, Goat, Human telomerase reverse transcriptase (hTERT), Immortalization

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Supplementary material

2017 Volume 63 Issue 1 Pages 27-36

DOI https://doi.org/10.1262/jrd.2016-111

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Abstract

With granulosa and theca cells, the ovaries are responsible for producing oocytes and secreting sex steroids such as estrogen and progesterone. Endoplasmic reticulum stress (ERS) plays an important role in follicle atresia and embryo implantation. In this study, goat granulosa cells were isolated from medium-sized (4–6 mm) healthy follicles. Primary granulosa cells were immortalized by transfection with human telomerase reverse transcriptase (hTERT) to establish a goat granulosa cell line (hTERT-GGCs). These hTERT-GGCs expressed hTERT and had relatively long telomeres at passage 50. Furthermore, hTERT-GGCs expressed the gonadotropin receptor genes CYP11A1, StAR, and CYP19A1, which are involved in steroidogenesis. Additionally, progesterone was detectable in hTERT-GGCs. Although the proliferation potential of hTERT-GGCs significantly improved, there was no evidence to suggest that the hTERT-GGCs are tumorigenic. In addition, thapsigargin (Tg) treatment led to a significant dose-dependent decrease in progesterone concentration and steroidogenic enzyme expression. In summary, we successfully generated a stable goat granulosa cell line. We found that Tg induced ERS in hTERT-GGCs, which reduced progesterone production and steroidogenic enzyme expression. Future studies may benefit from using this cell line as a model to explore the molecular mechanisms regulating steroidogenesis and apoptosis in goat granulosa cells.

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