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Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase

Reproduction is regulated by gonadotropins secreted from gonadotrophs. The production and secretion of gonadotropins are mainly regulated by gonadotropin-releasing hormone (GnRH). Agonists or antagonists that influence GnRH action on gonadotrophs are important to regulate reproduction; however, thes...

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Autores principales: SATOU, Kazuhiro, MOCHIMARU, Yuta, NAKAKURA, Takashi, KUSADA, Tomoyuki, NEGISHI, Jun, MUSHA, Shiori, YOSHIMURA, Nanaka, KATO, Yukio, TOMURA, Hideaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Society for Reproduction and Development 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5401814/
https://www.ncbi.nlm.nih.gov/pubmed/28090002
http://dx.doi.org/10.1262/jrd.2016-174
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author SATOU, Kazuhiro
MOCHIMARU, Yuta
NAKAKURA, Takashi
KUSADA, Tomoyuki
NEGISHI, Jun
MUSHA, Shiori
YOSHIMURA, Nanaka
KATO, Yukio
TOMURA, Hideaki
author_facet SATOU, Kazuhiro
MOCHIMARU, Yuta
NAKAKURA, Takashi
KUSADA, Tomoyuki
NEGISHI, Jun
MUSHA, Shiori
YOSHIMURA, Nanaka
KATO, Yukio
TOMURA, Hideaki
author_sort SATOU, Kazuhiro
collection PubMed
description Reproduction is regulated by gonadotropins secreted from gonadotrophs. The production and secretion of gonadotropins are mainly regulated by gonadotropin-releasing hormone (GnRH). Agonists or antagonists that influence GnRH action on gonadotrophs are important to regulate reproduction; however, these factors have not been fully characterized due to the lack of simple and easy-to-use techniques to detect gonadotropin secretion from gonadotropin-producing cells. In the present study, we found that Gaussia luciferase (Gluc), which was expressed in LβT2 cells, can be secreted like a luteinizing-hormone (LH) upon stimulation with GnRH. The Gluc secreted into the medium was easily monitored as luminescence signals. The detection range of the GnRH-induced Gluc activity was comparable to that of the enzyme-linked immunosorbent assay for LH. In addition, when the Gluc was expressed in AtT20 cells, which produce adrenocorticotropic hormone (ACTH), the Gluc activity in the medium increased in parallel with the ACTH secretion upon stimulation with corticotropin-releasing hormone. Thus, the Gluc assay in the present study can be easily used for high-throughput screening of factors that influence LH or ACTH secretion from LβT2 or AtT20 cells, respectively.
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spelling pubmed-54018142017-04-27 Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase SATOU, Kazuhiro MOCHIMARU, Yuta NAKAKURA, Takashi KUSADA, Tomoyuki NEGISHI, Jun MUSHA, Shiori YOSHIMURA, Nanaka KATO, Yukio TOMURA, Hideaki J Reprod Dev Technology Report Reproduction is regulated by gonadotropins secreted from gonadotrophs. The production and secretion of gonadotropins are mainly regulated by gonadotropin-releasing hormone (GnRH). Agonists or antagonists that influence GnRH action on gonadotrophs are important to regulate reproduction; however, these factors have not been fully characterized due to the lack of simple and easy-to-use techniques to detect gonadotropin secretion from gonadotropin-producing cells. In the present study, we found that Gaussia luciferase (Gluc), which was expressed in LβT2 cells, can be secreted like a luteinizing-hormone (LH) upon stimulation with GnRH. The Gluc secreted into the medium was easily monitored as luminescence signals. The detection range of the GnRH-induced Gluc activity was comparable to that of the enzyme-linked immunosorbent assay for LH. In addition, when the Gluc was expressed in AtT20 cells, which produce adrenocorticotropic hormone (ACTH), the Gluc activity in the medium increased in parallel with the ACTH secretion upon stimulation with corticotropin-releasing hormone. Thus, the Gluc assay in the present study can be easily used for high-throughput screening of factors that influence LH or ACTH secretion from LβT2 or AtT20 cells, respectively. The Society for Reproduction and Development 2017-01-15 2017-04 /pmc/articles/PMC5401814/ /pubmed/28090002 http://dx.doi.org/10.1262/jrd.2016-174 Text en ©2017 Society for Reproduction and Development This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Technology Report
SATOU, Kazuhiro
MOCHIMARU, Yuta
NAKAKURA, Takashi
KUSADA, Tomoyuki
NEGISHI, Jun
MUSHA, Shiori
YOSHIMURA, Nanaka
KATO, Yukio
TOMURA, Hideaki
Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title_full Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title_fullStr Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title_full_unstemmed Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title_short Easy detection of hormone secretion from LβT2 cells by using Gaussia luciferase
title_sort easy detection of hormone secretion from lβt2 cells by using gaussia luciferase
topic Technology Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5401814/
https://www.ncbi.nlm.nih.gov/pubmed/28090002
http://dx.doi.org/10.1262/jrd.2016-174
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