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Construction and expression of vectors encoding biologically active rodent gonadotropins

The gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are important hormones in vertebrate reproduction. The isolation of gonadotropins from the pituitary gland is sub-optimal, as the cross-contamination of one hormone with another is common and often results in the var...

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Autores principales: OHTA, Akihiko, TSUNODA, Yuichiro, TAMURA, Yoshihiko, IINO, Kayoko, NISHIMURA, Naoto, NISHIHARA, Hiroto, TAKANASHI, Haruka, YOSHIDA, Saishu, KATO, Takako, KATO, Yukio
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/PMC5735272/
https://www.ncbi.nlm.nih.gov/pubmed/29033405
http://dx.doi.org/10.1262/jrd.2017-091
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author OHTA, Akihiko
TSUNODA, Yuichiro
TAMURA, Yoshihiko
IINO, Kayoko
NISHIMURA, Naoto
NISHIHARA, Hiroto
TAKANASHI, Haruka
YOSHIDA, Saishu
KATO, Takako
KATO, Yukio
author_facet OHTA, Akihiko
TSUNODA, Yuichiro
TAMURA, Yoshihiko
IINO, Kayoko
NISHIMURA, Naoto
NISHIHARA, Hiroto
TAKANASHI, Haruka
YOSHIDA, Saishu
KATO, Takako
KATO, Yukio
author_sort OHTA, Akihiko
collection PubMed
description The gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are important hormones in vertebrate reproduction. The isolation of gonadotropins from the pituitary gland is sub-optimal, as the cross-contamination of one hormone with another is common and often results in the variation in the measured activity of LH and FSH. The production of recombinant hormones is, therefore, a viable approach to solve this problem. This study aimed to express recombinant rat, mouse, and mastomys FSH and LH in Chinese hamster ovary (CHO) cells. Their common α-subunits along with their hormone-specific β-subunits were encoded in a single mammalian expression vector. FSH from all three species was expressed, whereas expression was achieved only for the mouse LH. Immunohistochemistry for rat alpha subunit of glycoprotein hormone (αGSU) and LHβ and FSHβ subunits confirmed the production of the dimeric hormone in CHO cells. The recombinant rodent gonadotropins were confirmed to be biologically active; estradiol production was increased by recombinant FSH in granulosa cells, while recombinant LH increased testosterone production in Leydig cells.
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spelling pubmed-57352722017-12-21 Construction and expression of vectors encoding biologically active rodent gonadotropins OHTA, Akihiko TSUNODA, Yuichiro TAMURA, Yoshihiko IINO, Kayoko NISHIMURA, Naoto NISHIHARA, Hiroto TAKANASHI, Haruka YOSHIDA, Saishu KATO, Takako KATO, Yukio J Reprod Dev Technology Report The gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are important hormones in vertebrate reproduction. The isolation of gonadotropins from the pituitary gland is sub-optimal, as the cross-contamination of one hormone with another is common and often results in the variation in the measured activity of LH and FSH. The production of recombinant hormones is, therefore, a viable approach to solve this problem. This study aimed to express recombinant rat, mouse, and mastomys FSH and LH in Chinese hamster ovary (CHO) cells. Their common α-subunits along with their hormone-specific β-subunits were encoded in a single mammalian expression vector. FSH from all three species was expressed, whereas expression was achieved only for the mouse LH. Immunohistochemistry for rat alpha subunit of glycoprotein hormone (αGSU) and LHβ and FSHβ subunits confirmed the production of the dimeric hormone in CHO cells. The recombinant rodent gonadotropins were confirmed to be biologically active; estradiol production was increased by recombinant FSH in granulosa cells, while recombinant LH increased testosterone production in Leydig cells. The Society for Reproduction and Development 2017-10-13 2017-12 /pmc/articles/PMC5735272/ /pubmed/29033405 http://dx.doi.org/10.1262/jrd.2017-091 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
OHTA, Akihiko
TSUNODA, Yuichiro
TAMURA, Yoshihiko
IINO, Kayoko
NISHIMURA, Naoto
NISHIHARA, Hiroto
TAKANASHI, Haruka
YOSHIDA, Saishu
KATO, Takako
KATO, Yukio
Construction and expression of vectors encoding biologically active rodent gonadotropins
title Construction and expression of vectors encoding biologically active rodent gonadotropins
title_full Construction and expression of vectors encoding biologically active rodent gonadotropins
title_fullStr Construction and expression of vectors encoding biologically active rodent gonadotropins
title_full_unstemmed Construction and expression of vectors encoding biologically active rodent gonadotropins
title_short Construction and expression of vectors encoding biologically active rodent gonadotropins
title_sort construction and expression of vectors encoding biologically active rodent gonadotropins
topic Technology Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5735272/
https://www.ncbi.nlm.nih.gov/pubmed/29033405
http://dx.doi.org/10.1262/jrd.2017-091
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