Reversible disassembly of the actin cytoskeleton improves the survival rate and developmental competence of cryopreserved mouse oocytes.
Effective cryopreservation of oocytes is critically needed in many areas of human reproductive medicine and basic science, such as stem cell research. Currently, oocyte cryopreservation has a low success rate. The goal of this study was to understand the mechanisms associated with oocyte cryopreservation through biophysical means using a mouse model. Specifically, we experimentally investigated the biomechanical properties of the ooplasm prior and after cryopreservation as well as the consequences of reversible dismantling of the F-actin network in mouse oocytes prior to freezing. The study was complemented with the evaluation of post-thaw developmental competence of oocytes after in vitro fertilization. Our results show that the freezing-thawing process markedly alters the physiological viscoelastic properties of the actin cytoskeleton. The reversible depolymerization of the F-actin network prior to freezing preserves normal ooplasm viscoelastic properties, results in high post-thaw survival and significantly improves developmental competence. These findings provide new information on the biophysical characteristics of mammalian oocytes, identify a pathophysiological mechanism underlying cryodamage and suggest a novel cryopreservation method.
Hosu
Basarab G
BG
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America.
Mullen
Steven F
SF
Critser
John K
JK
Forgacs
Gabor
G
D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D004548:Elasticity; D005260:Female; D005307:Fertilization in Vitro; D008280:Magnetics; D051379:Mice; D008954:Models, Biological; D008962:Models, Theoretical; D009865:Oocytes / Q000378:metabolism*; D013075:Sperm Capacitation
D000199:Actins; D003451:Cryoprotective Agents