RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts

Abstract Embryonic cryopreservation has a relatively low survival rate because of cytoskeletal damage. However, molecular anti-freezing mechanisms have been largely unexplored. This study investigated the significance of RhoA, involved in embryonic development, and the Rho/RhoA-associated kinase (RO...

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Autores principales:	Meichao Gu, Hemin Ni, Xihui Sheng, Alfredo Pauciullo, Yunhai Liu, Yong Guo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9768612cf7284f5eaf4556c5130a982d
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spelling	oai:doaj.org-article:9768612cf7284f5eaf4556c5130a982d2021-12-02T16:06:44ZRhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts10.1038/s41598-017-07066-22045-2322https://doaj.org/article/9768612cf7284f5eaf4556c5130a982d2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07066-2https://doaj.org/toc/2045-2322Abstract Embryonic cryopreservation has a relatively low survival rate because of cytoskeletal damage. However, molecular anti-freezing mechanisms have been largely unexplored. This study investigated the significance of RhoA, involved in embryonic development, and the Rho/RhoA-associated kinase (ROCK) signalling pathway in cryopreservation. The anti-freezing mechanism in murine dormant embryos, compared with normal blastocysts, was assessed by combining molecular, physiological and pharmacological approaches. Real-time PCR and western blotting experiments showed high RhoA expression in cryo-dormant and dormant embryos. RhoA GTPases were overexpressed on the surface of trophectoderm cells in dormant embryos. Treatment with Y-27632, a ROCK antagonist, decreased survival of both normal and dormant blastocysts, while recombinant RhoA protein remarkably increased survival, after freeze–thawing, of normal hatched blastocysts. Our findings elucidated the molecular mechanism of anti-freezing, involving RhoA phosphorylation, meditated by the Rho/ROCK signalling pathway, in hatched and diapaused murine blastocysts. In addition, evidence for a potentially protective additive suggests a new method for improving the anti-freezing potential of mammalian embryos, without protecting the zona pellucida.Meichao GuHemin NiXihui ShengAlfredo PauciulloYunhai LiuYong GuoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-7 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Meichao Gu Hemin Ni Xihui Sheng Alfredo Pauciullo Yunhai Liu Yong Guo RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
description	Abstract Embryonic cryopreservation has a relatively low survival rate because of cytoskeletal damage. However, molecular anti-freezing mechanisms have been largely unexplored. This study investigated the significance of RhoA, involved in embryonic development, and the Rho/RhoA-associated kinase (ROCK) signalling pathway in cryopreservation. The anti-freezing mechanism in murine dormant embryos, compared with normal blastocysts, was assessed by combining molecular, physiological and pharmacological approaches. Real-time PCR and western blotting experiments showed high RhoA expression in cryo-dormant and dormant embryos. RhoA GTPases were overexpressed on the surface of trophectoderm cells in dormant embryos. Treatment with Y-27632, a ROCK antagonist, decreased survival of both normal and dormant blastocysts, while recombinant RhoA protein remarkably increased survival, after freeze–thawing, of normal hatched blastocysts. Our findings elucidated the molecular mechanism of anti-freezing, involving RhoA phosphorylation, meditated by the Rho/ROCK signalling pathway, in hatched and diapaused murine blastocysts. In addition, evidence for a potentially protective additive suggests a new method for improving the anti-freezing potential of mammalian embryos, without protecting the zona pellucida.
format	article
author	Meichao Gu Hemin Ni Xihui Sheng Alfredo Pauciullo Yunhai Liu Yong Guo
author_facet	Meichao Gu Hemin Ni Xihui Sheng Alfredo Pauciullo Yunhai Liu Yong Guo
author_sort	Meichao Gu
title	RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
title_short	RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
title_full	RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
title_fullStr	RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
title_full_unstemmed	RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
title_sort	rhoa phosphorylation mediated by rho/rhoa-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/9768612cf7284f5eaf4556c5130a982d
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RhoA phosphorylation mediated by Rho/RhoA-associated kinase pathway improves the anti-freezing potentiality of murine hatched and diapaused blastocysts

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