In vitro fertilization of in vitro matured canine oocytes using frozen-thawed dog semen.
Experiments were conducted to evaluate in vitro fertilization (IVF) of in vitro matured (IVM) bitch oocytes using dog spermatozoa frozen in three different extenders. Sperm-rich fraction from eight ejaculates of five dogs was frozen in each one of three egg yolk Tris extenders with additional: (A) 1.4 g citric acid and 0.8 g glucose; (B) 0.7 g citric acid and 3.5 g glucose; or (C) 1.4 g citric acid and 0.8 g fructose (all with 5% glycerol in 100 mL milliQ water). Thawed sperm were co-incubated with IVM bitch oocytes for 6 h. Oocytes were fixed and evaluated under an epifluorescence microscope; penetrated oocytes were defined as those having sperm heads in the perivitelline space or in the oocyte cytoplasm. Higher penetration rates (P < 0.05) were obtained in oocytes cultured with spermatozoa frozen in extenders B and C than those frozen in extender A (33.1, 34.2 and 26.4%, respectively).
De los Reyes
M
M
Laboratory of Animal Reproduction, Faculty of Veterinary Sciences, Casilla 2 Correo 15, Santiago, Chile. mdlreyes@uchile.cl
Carrion
R
R
Barros
C
C
D000818:Animals; D015925:Cryopreservation / Q000662:veterinary*; D004285:Dogs / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D005456:Fluorescent Dyes / Q000737:chemistry; D008297:Male; D008856:Microscopy, Fluorescence / Q000662:veterinary; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D011419:Propidium / Q000737:chemistry; D012662:Semen Preservation / Q000662:veterinary*; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*
D005456:Fluorescent Dyes; D011419:Propidium