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BOR - Papers in Press, published online ahead of print August 18, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.031989
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Submitted May 13, 2004
Returned for revision June 29, 2004
Accepted August 11, 2004

Gamete Biology


Sphingosine 1-Phosphate Protects Bovine Oocytes from Heat Shock During Maturation

Z. Roth and P. J. Hansen *

* To whom correspondence should be addressed. E-mail: hansen{at}animal.ufl.edu.

Abstract
Sphingosine 1-phosphate (S1P) is a sphingolipid metabolite that can block apoptosis by counteracting the pro-apoptotic effects of ceramide. Experiments were performed to evaluate whether S1P blocks the disruption in oocyte developmental competence caused by heat shock. Cumulus-oocyte complexes (COCs) were placed in maturation medium and cultured at 38.5°C or 41°C for the first 12 h of maturation. Incubation during the last 10 h of maturation, fertilization and embryonic development were performed at 38.5°C. Heat shock during the first 12 h of maturation reduced cleavage rate, the number of oocytes developing to the blastocyst stage and the percentage of cleaved embryo that subsequently developed to blastocysts. Addition of 50 nM S1P to maturation medium had no effect on oocytes matured at 38.5°C but blocked effects of thermal stress on cleavage and subsequent development. The blastocysts formed at day 8 did not differ between S1P and control groups in caspase activity, total cell number, or percentage of cells that were apoptotic. Blocking endogenous generation of S1P by addition of 50 nM N1N-dimethylsphingosine, a sphingosine kinase inhibitor, reduced or tended to reduce cleavage rate and blastocyst development regardless of whether maturation of COCs was at 38.5 or 41°C. Results demonstrate that S1P protects oocytes from a physiologically-relevant heat shock and affects oocyte maturation even in the absence of heat shock. The S1P-treated oocytes that survived heat shock and became blastocysts had a normal developmental potential as determined by caspase activity, total cell number and percentage of apoptotic cells. Thus, modulation of developmental competence of oocytes using S1P may be a useful approach for enhancing fertility in situations where developmental competence of oocytes is compromised.

Key words: Embryo • Environment • Gamete Biology • Early development • Oocyte development


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