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Fluorescence-based visualization of autophagic activity predicts mouse embryo viability

Embryo quality is a critical parameter in assisted reproductive technologies. Although embryo quality can be evaluated morphologically, embryo morphology does not correlate perfectly with embryo viability. To improve this, it is important to understand which molecular mechanisms are involved in embr...

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Autores principales: Tsukamoto, Satoshi, Hara, Taichi, Yamamoto, Atsushi, Kito, Seiji, Minami, Naojiro, Kubota, Toshiro, Sato, Ken, Kokubo, Toshiaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3970120/
https://www.ncbi.nlm.nih.gov/pubmed/24681842
http://dx.doi.org/10.1038/srep04533
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author Tsukamoto, Satoshi
Hara, Taichi
Yamamoto, Atsushi
Kito, Seiji
Minami, Naojiro
Kubota, Toshiro
Sato, Ken
Kokubo, Toshiaki
author_facet Tsukamoto, Satoshi
Hara, Taichi
Yamamoto, Atsushi
Kito, Seiji
Minami, Naojiro
Kubota, Toshiro
Sato, Ken
Kokubo, Toshiaki
author_sort Tsukamoto, Satoshi
collection PubMed
description Embryo quality is a critical parameter in assisted reproductive technologies. Although embryo quality can be evaluated morphologically, embryo morphology does not correlate perfectly with embryo viability. To improve this, it is important to understand which molecular mechanisms are involved in embryo quality control. Autophagy is an evolutionarily conserved catabolic process in which cytoplasmic materials sequestered by autophagosomes are degraded in lysosomes. We previously demonstrated that autophagy is highly activated after fertilization and is essential for further embryonic development. Here, we developed a simple fluorescence-based method for visualizing autophagic activity in live mouse embryos. Our method is based on imaging of the fluorescence intensity of GFP-LC3, a versatile marker for autophagy, which is microinjected into the embryos. Using this method, we show that embryonic autophagic activity declines with advancing maternal age, probably due to a decline in the activity of lysosomal hydrolases. We also demonstrate that embryonic autophagic activity is associated with the developmental viability of the embryo. Our results suggest that embryonic autophagic activity can be utilized as a novel indicator of embryo quality.
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spelling pubmed-39701202014-04-01 Fluorescence-based visualization of autophagic activity predicts mouse embryo viability Tsukamoto, Satoshi Hara, Taichi Yamamoto, Atsushi Kito, Seiji Minami, Naojiro Kubota, Toshiro Sato, Ken Kokubo, Toshiaki Sci Rep Article Embryo quality is a critical parameter in assisted reproductive technologies. Although embryo quality can be evaluated morphologically, embryo morphology does not correlate perfectly with embryo viability. To improve this, it is important to understand which molecular mechanisms are involved in embryo quality control. Autophagy is an evolutionarily conserved catabolic process in which cytoplasmic materials sequestered by autophagosomes are degraded in lysosomes. We previously demonstrated that autophagy is highly activated after fertilization and is essential for further embryonic development. Here, we developed a simple fluorescence-based method for visualizing autophagic activity in live mouse embryos. Our method is based on imaging of the fluorescence intensity of GFP-LC3, a versatile marker for autophagy, which is microinjected into the embryos. Using this method, we show that embryonic autophagic activity declines with advancing maternal age, probably due to a decline in the activity of lysosomal hydrolases. We also demonstrate that embryonic autophagic activity is associated with the developmental viability of the embryo. Our results suggest that embryonic autophagic activity can be utilized as a novel indicator of embryo quality. Nature Publishing Group 2014-03-31 /pmc/articles/PMC3970120/ /pubmed/24681842 http://dx.doi.org/10.1038/srep04533 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images in this article are included in the article's Creative Commons license, unless indicated otherwise in the image credit; if the image is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the image. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Article
Tsukamoto, Satoshi
Hara, Taichi
Yamamoto, Atsushi
Kito, Seiji
Minami, Naojiro
Kubota, Toshiro
Sato, Ken
Kokubo, Toshiaki
Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title_full Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title_fullStr Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title_full_unstemmed Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title_short Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
title_sort fluorescence-based visualization of autophagic activity predicts mouse embryo viability
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3970120/
https://www.ncbi.nlm.nih.gov/pubmed/24681842
http://dx.doi.org/10.1038/srep04533
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