Role of kisspeptin on cell proliferation and steroidogenesis in luteal cells in vitro and in vivo

Chi-Ming Chiang; Hsin-Yi Chiu; Chia-Sheng Chang; Ya-Yun Chien; De-Shien Jong; Leang-Shin Wu; Chih-Hsien Chiu

doi:10.1097/JCMA.0000000000000508

Role of kisspeptin on cell proliferation and steroidogenesis in luteal cells in vitro and in vivo

J Chin Med Assoc. 2021 Apr 1;84(4):389-399. doi: 10.1097/JCMA.0000000000000508.

Authors

Chi-Ming Chiang^{1

2

3}, Hsin-Yi Chiu^{1

4

5

6

7}, Chia-Sheng Chang¹, Ya-Yun Chien¹, De-Shien Jong¹, Leang-Shin Wu¹, Chih-Hsien Chiu¹

Affiliations

¹ Department of Animal Science and Technology, National Taiwan University, Taipei, Taiwan, ROC.
² Department of Orthopedics Surgery, Cardinal Tien Hospital, New Taipei City, Taiwan, ROC.
³ Professional Master Program in Artificial Intelligence in Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan, ROC.
⁴ Division of Thoracic Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei, Taiwan, ROC.
⁵ Department of Medical Education, Taipei Medical University Hospital, Taipei, Taiwan, ROC.
⁶ Department of Education and Humanities in Medicine, School of Medicine, Taipei Medical University, Taipei, Taiwan, ROC.
⁷ Department of Surgery, School of Medicine, Taipei Medical University, Taipei, Taiwan, ROC.

PMID: 33784266
DOI: 10.1097/JCMA.0000000000000508

Abstract

Background: Kisspeptin (KISS1) and kisspeptin receptor (KISS1R) are essential gatekeepers of the reproductive system. The functions of KISS1 and KISS1R in corpus luteal cells remain ambiguous. The objective was to observe normal physiologic functions of corpus luteal cells in vivo and clarify the functions of KISS1 in vitro.

Methods: We conducted an in vivo observation of cellular patterns as well as the levels of steroidogenic enzymes and KISS1/KISS1R in corpus luteal cells obtained from female crossbred Taiwan native goats in the estrous cycle; the observation was performed using hematoxylin and eosin and immunohistochemistry staining. Subsequently, we used kisspeptin-10 (Kp-10) to stimulate temperature sensitive-caprine luteal cell line (ts-CLC-D) cells to investigate the progesterone (P4) levels, steroidogenic messenger RNA (mRNA)/protein levels, cell survival rate, intracellular Ca2+ concentration, and cell proliferation-related mRNA/protein levels in the mitogen-activated protein kinase pathway in vitro by applying immunofluorescence staining, Western blotting, 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay, and real-time polymerase chain reaction.

Results: We observed the presence of proteins and mRNAs for STAR, CYP11A1, HSD3B, KISS1, and KISS1R in the corpus luteal cells from goats in vivo. In vitro, the addition of Kp-10 reduced the P4 levels (p < 0.01) and increased cell proliferation (p < 0.05) of the ts-CLC-D cells. Furthermore, we found that the levels of proteins and mRNA for STAR, CYP11A1, and HSD3B decreased significantly when Kp-10 was added (p < 0.05). However, adding Kp-10 did not affect the mRNA levels for PLCG2, DAG1, PRKCA, KRAS, RAF1, MAP2K1, MAP2K2, MAPK3, MAPK1, and MAPK14.

Conclusion: We determined that KISS1 could affect the P4 levels, steroidogenesis, and cell proliferation in luteal cells. However, further research is required to clarify how KISS1 regulates proliferation and steroid production in luteal cells.

MeSH terms

Animals
Cell Proliferation / drug effects*
Cell Survival
Female
Gene Expression / genetics
Goats
Kisspeptins / pharmacology*
Luteal Cells / drug effects*
Polymerase Chain Reaction
RNA, Messenger
Taiwan

Substances

Kisspeptins
RNA, Messenger