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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Society for Reproduction and Development
2017
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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. |
format | Online Article Text |
id | pubmed-5735272 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The Society for Reproduction and Development |
record_format | MEDLINE/PubMed |
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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