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雌激素增加了新生成的源自山羊前脑区的永生化 KISS1 表达细胞系中 KISS1 的表达。

Estrogen increases KISS1 expression in newly generated immortalized KISS1-expressing cell line derived from goat preoptic area.

机构信息

Laboratory of Theriogenology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.

Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.

出版信息

J Reprod Dev. 2021 Feb 15;67(1):15-23. doi: 10.1262/jrd.2020-053. Epub 2020 Oct 25.

DOI:10.1262/jrd.2020-053
PMID:33100283
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7902218/
Abstract

Kisspeptin neurons located in the hypothalamic preoptic area (POA) are suggested to be responsible for the induction of the gonadotropin-releasing hormone (GnRH) surge and the following luteinizing hormone (LH) surge to regulate female mammals' ovulation. Accumulating evidence demonstrates that the preovulatory level of estrogen activates the POA kisspeptin neurons (estrogen positive feedback), which in turn induces a GnRH/LH surge. This study aimed to derive a cell line from goat POA kisspeptin neurons as an in vitro model to analyze the estrogen positive feedback mechanism in ruminants. Neuron-derived cell clones obtained by the immortalization of POA tissue from a female Shiba goat fetus were analyzed for the expression of kisspeptin (KISS1) and estrogen receptor α (ESR1) genes using quantitative real-time reverse transcription-polymerase chain reaction and three cell clones were selected as POA kisspeptin neuron cell line candidates. One cell line (GP64) out of the three clones showed significant increase in the KISS1 level by incubation with estradiol for 24 h, indicating that the GP64 cells mimic endogenous goat POA kisspeptin neurons. The GP64 cells showed immunoreactivities for kisspeptin and estrogen receptor α and retained a stable growth rate throughout three passages. Further, intracellular calcium levels in the GP64 cells were increased by the KCl challenge, indicating their neurosecretory ability. In conclusion, we generated a new KISS1-expressing cell line derived from goat POA. The current GP64 cell line could be a useful model to elucidate the estrogen positive feedback mechanism responsible for the GnRH/LH surge generation in ruminants.

摘要

位于下丘脑前脑区（POA）的 Kisspeptin 神经元被认为负责诱导促性腺激素释放激素（GnRH）激增和随后的黄体生成素（LH）激增，以调节雌性哺乳动物的排卵。越来越多的证据表明，促排卵前的雌激素水平激活了 POA Kisspeptin 神经元（雌激素正反馈），进而诱导 GnRH/LH 激增。本研究旨在从山羊 POA Kisspeptin 神经元中获得细胞系，作为分析反刍动物中雌激素正反馈机制的体外模型。通过对雌性柴犬胎儿 POA 组织的永生化获得神经元衍生细胞克隆，使用定量实时逆转录-聚合酶链反应分析 Kisspeptin（KISS1）和雌激素受体α（ESR1）基因的表达，选择了 3 个细胞克隆作为 POA Kisspeptin 神经元细胞系候选物。其中一个细胞系（GP64）在孵育 24 小时后，通过与雌二醇孵育显示 KISS1 水平显著增加，表明 GP64 细胞模拟内源性山羊 POA Kisspeptin 神经元。GP64 细胞对 Kisspeptin 和雌激素受体 α 表现出免疫反应性，并在整个 3 个传代过程中保持稳定的增长率。此外，GP64 细胞中的细胞内钙水平通过 KCl 挑战增加，表明它们具有神经分泌能力。总之，我们从山羊 POA 中产生了一种新的 KISS1 表达细胞系。目前的 GP64 细胞系可以作为阐明负责反刍动物 GnRH/LH 激增产生的雌激素正反馈机制的有用模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/d36973655420/jrd-67-015-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/4e3765b192a2/jrd-67-015-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/e1106253bdc7/jrd-67-015-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/cc0eda1dfa2d/jrd-67-015-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/7f6a07caf289/jrd-67-015-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/d36973655420/jrd-67-015-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/4e3765b192a2/jrd-67-015-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/e1106253bdc7/jrd-67-015-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/cc0eda1dfa2d/jrd-67-015-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/7f6a07caf289/jrd-67-015-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb3e/7902218/d36973655420/jrd-67-015-g005.jpg

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J Neuroendocrinol. 2020 Jun;32(6):e12857. doi: 10.1111/jne.12857. Epub 2020 May 20.
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Sexually dimorphic gene expression and neurite sensitivity to estradiol in fetal arcuate Kiss1 cells.胎儿弓状核 Kiss1 细胞中存在性别二态性基因表达和雌激素对轴突敏感性。
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Role of kisspeptin neurons as a GnRH surge generator: Comparative aspects in rodents and non-rodent mammals.
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