Effect of 1,2-propanediol versus 1,2-ethanediol on subsequent oocyte maturation, spindle integrity, fertilization, and embryo development in vitro in the domestic cat.
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 degrees C) or 0 degrees 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 degrees 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 degrees 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 degrees C) or not (1.5 M PrOH at 25 degrees 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 degrees C, a treatment that still allowed approximately 60% of the oocytes to reach MII and approximately 20% to form blastocysts.
Comizzoli
Pierre
P
Department of Reproductive Sciences, Smithsonian's National Zoological Park, 3001 Connecticut Avenue NW, Washington, D.C. 20008, USA. comizzolip@si.edu
Wildt
David E
DE
Pukazhenthi
Budhan S
BS
D000818:Animals; D002415:Cats; D002467:Cell Nucleus / Q000187:drug effects; D003451:Cryoprotective Agents / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects*; D019855:Ethylene Glycol / Q000494:pharmacology*; D005306:Fertilization / Q000187:drug effects; D005307:Fertilization in Vitro; D066298:In Vitro Techniques; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D019946:Propylene Glycol / Q000494:pharmacology*; D008941:Spindle Apparatus / Q000187:drug effects
D003451:Cryoprotective Agents; D019946:Propylene Glycol; D019855:Ethylene Glycol