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Mutual Interactions Between GnRH and Kisspeptin in GnRH- and Kiss-1-Expressing Immortalized Hypothalamic Cell Models

  • Reproductive Endocrinology; Original Article
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Abstract

Kisspeptin and gonadotropin-releasing hormone (GnRH) are central regulators of the hypothalamic-pituitary-gonadal axis and control female reproductive functions. Recently established mHypoA-50 and mHypoA-55 cells are immortalized hypothalamic neuronal cell models that originated from the anteroventral periventricular nucleus (AVPV) and arcuate nucleus (ARC) regions of the mouse hypothalamus, respectively. mHypoA-50 or mHypoA-55 cells were stimulated with kisspeptin-10 (KP10) and GnRH, after which the expression of kisspeptin and GnRH was determined. Primary cultures of fetal rat brain cells were also examined. mHypoA-50 and mHypoA-55 cells expressed mRNA for Kiss-1 (which encodes kisspeptin) and GnRH as well as receptors for kisspeptin and GnRH. We found that Kiss-1 mRNA expression was significantly increased in mHypoA-50 AVPV cells by KP10 and GnRH stimulation. Kisspeptin protein expression was also increased by KP10 and GnRH stimulation in these cells. In contrast, GnRH expression was unchanged in mHypoA-50 AVPV cells by KP10 and GnRH stimulation. In mHypoA-55 ARC cells, kisspeptin expression was also significantly increased at the mRNA and protein levels by KP10 and GnRH stimulation; however, GnRH expression was also upregulated by KP10 and GnRH stimulation in these cells. KP10 and estradiol (E2) both increased Kiss-1 gene expression in mHypoA-50 AVPV cells, but combined stimulation with KP10 and E2 did not potentiate their individual effects on Kiss-1 gene expression. On the other hand, E2 did not increase Kiss-1 gene expression in mHypoA-55 ARC cells, and the KP10-induced increase of Kiss-1 gene expression was inhibited in the presence of E2 in these cells. KP10 and GnRH significantly increased c-Fos protein expression in the mHypoA-50 AVPV and mHypoA-55 ARC cell lines. In primary cultures of fetal rat neuronal cells, KP10 significantly increased Kiss-1 gene expression, whereas GnRH significantly increased GnRH gene expression. We found that kisspeptin and GnRH affected Kiss-1- and GnRH-expressing hypothalamic cells and modulated Kiss-1 and/or GnRH gene expression with a concomitant increase in c-Fos protein expression. A mutual- or self-regulatory system might be present in Kiss-1 and/or GnRH neurons in the hypothalamus.

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Abbreviations

ARC:

arcuate nucleus

AVPV:

anteroventral periventricular nucleus

GnRH:

gonadotropin-releasing hormone

GnRHR:

gonadotropin-releasing hormone receptor

Kiss1R:

kisspeptin receptor

KNDy:

kisspeptin-neurokinin B-dynorphin A

KP10:

kisspeptin-10

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Funding

This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan (to HK and AO).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, School of Medicine, Shimane University, 89-1 Enya-cho, Izumo, Shimane, 693-8501, Japan

    Haruhiko Kanasaki, Tuvshintugs Tumurbaatar, Zolzaya Tumurgan, Aki Oride, Hiroe Okada & Satoru Kyo

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  1. Haruhiko Kanasaki
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  2. Tuvshintugs Tumurbaatar
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  3. Zolzaya Tumurgan
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  4. Aki Oride
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  6. Satoru Kyo
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Contributions

HK and SA conceived and designed the experiments. HK, TT, ZT, AO, and HO performed the experiments. HK wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Haruhiko Kanasaki.

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The study protocol was approved by the committee of the Experimental Animal Center for Integrated Research, Shimane University. Consent to participate is not applicable in this study.

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Kanasaki, H., Tumurbaatar, T., Tumurgan, Z. et al. Mutual Interactions Between GnRH and Kisspeptin in GnRH- and Kiss-1-Expressing Immortalized Hypothalamic Cell Models. Reprod. Sci. 28, 3380–3389 (2021). https://doi.org/10.1007/s43032-021-00695-z

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