Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print April 14, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.027920
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BIOLOGY OF REPRODUCTION 71, 598–604 (2004)
DOI: 10.1095/biolreprod.104.027920
© 2004 by the Society for the Study of Reproduction, Inc.

Reproductive Technology

Effect of 1,2-Propanediol Versus 1,2-Ethanediol on Subsequent Oocyte Maturation, Spindle Integrity, Fertilization, and Embryo Development In Vitro in the Domestic Cat1

Pierre Comizzoli2, David E. Wildt, and Budhan S. Pukazhenthi

Department of Reproductive Sciences, Smithsonian's National Zoological Park, Washington, D.C. 20008

This study assessed the impact of various cryoprotectant (CPA) exposures on nuclear and cytoplasmic maturation in the immature cat oocyte as a prerequisite to formulating a successful cryopreservation protocol. In experiment 1, immature oocytes were exposed to 0, 0.75, 1.5, or 3.0 M of 1,2-propanediol (PrOH) or 1,2-ethanediol (EG) at room temperature (25°C) or 0°C for 30 min. After CPA removal and in vitro maturation, percentage of oocytes reaching metaphase II (MII) was reduced after exposure to 3.0 M PrOH at 0°C or 3.0 M EG at both temperatures. All CPA exposures increased MII spindle abnormalities compared to control, except 1.5 M PrOH at 25°C. In experiments 2 and 3, immature oocytes were exposed to CPA conditions yielding optimal nuclear maturation that either had caused spindle damage (0.75 M PrOH, 1.5 M EG, and 3.0 M PrOH at 25°C) or not (1.5 M PrOH at 25°C). After maturation and insemination in vitro, oocytes were cultured for 7 days to assess treatment influence on developmental competence. CPA exposure did not affect fertilization, but the high incidence of MII spindle abnormalities resulted in a low percentage of cleaved embryos. Blastocyst formation and quality were influenced by both CPA types (EG was more detrimental than PrOH) and concentration (3.0 M was more detrimental than 1.5 M). Overall, cat oocytes appear to be highly sensitive to CPA except after exposure to 1.5 M PrOH at 25°C, a treatment that still allowed ~60% of the oocytes to reach MII and ~20% to form blastocysts.

1 Supported by a fellowship to P.C. from the Smithsonian Institution Scholarly Studies Program and the National Institutes of Health (KO1 RR00135).

2 Correspondence: Pierre Comizzoli, Department of Reproductive Sciences, Smithsonian's National Zoological Park, 3001 Connecticut Ave. NW, Washington, DC 20008-2598. FAX: 202 673 4733; comizzolip{at}si.edu






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Copyright © 2004 by the Society for the Study of Reproduction.