Biology of Reproduction, Vol 50, 510-515, Copyright © 1994 by Society for the Study of Reproduction

ARTICLES

Penetration of porcine oocytes during maturation in vitro by cryopreserved, ejaculated spermatozoa

WH Wang, LR Abeydeera, K Okuda and K Niwa
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Japan.

The present study was conducted to examine the penetrability in vitro of immature porcine oocytes with or without cumulus cells. Porcine oocytes were cultured for 0-36 h, at 39 degrees C in 5% CO2 in air, in modified tissue culture medium 199 (TCM-199B at pH 7.4) supplemented with 10 IU eCG/ml, 10 IU hCG/ml, and 1 microgram estradiol-17 beta/ml. At various times after the beginning of culture, some oocytes were freed from the cumulus (cumulus and corona cells), and cumulus-intact or cumulus-free oocytes were inseminated with cryopreserved ejaculated spermatozoa in TCM-199B (pH 7.8) containing 5 mM caffeine. When cumulus- free oocytes were examined 14 h after insemination, high proportions (69-84%) were penetrated and there were no significant differences among different periods of maturation culture. The incidence (47-68%) of polyspermy and the number (1.7-3.1) of spermatozoa that penetrated per oocyte were also not significantly different among oocytes cultured for 0-36 h. In cumulus-intact oocytes, however, the first evidence of penetration (15%) was observed in oocytes cultured for 6 h. The penetration rates increased significantly as the period of culture was prolonged up to 24 h. Almost all (95-100%) oocytes were penetrated when they were inseminated 24-36 h after the beginning of culture, by which time the cumulus masses showed moderate to complete expansion except for the corona radiata. A similar correlation was also observed for incidence of polyspermy and number of spermatozoa penetrated per oocyte. The presence of well-expanded cumulus around the oocyte during fertilization promoted male pronuclear formation in penetrant oocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

				Y.-J. Yi, G. Manandhar, M. Sutovsky, R. Li, V. Jonakova, R. Oko, C.-S. Park, R. S Prather, and P. Sutovsky Ubiquitin C-Terminal Hydrolase-Activity Is Involved in Sperm Acrosomal Function and Anti-Polyspermy Defense During Porcine Fertilization Biol Reprod, November 1, 2007; 77(5): 780 - 793. [Abstract] [Full Text] [PDF]

				W. Ma, D. Zhang, Y. Hou, Y.-H. Li, Q.-Y. Sun, X.-F. Sun, and W.-H. Wang Reduced Expression of MAD2, BCL2, and MAP Kinase Activity in Pig Oocytes after In Vitro Aging Are Associated with Defects in Sister Chromatid Segregation During Meiosis II and Embryo Fragmentation After Activation Biol Reprod, February 1, 2005; 72(2): 373 - 383. [Abstract] [Full Text] [PDF]

				H. Tatemoto, N. Muto, S.-D. Yim, and T. Nakada Anti-Hyaluronidase Oligosaccharide Derived from Chondroitin Sulfate A Effectively Reduces Polyspermy During In Vitro Fertilization of Porcine Oocytes Biol Reprod, January 1, 2005; 72(1): 127 - 134. [Abstract] [Full Text] [PDF]

				D. Zhang, W. Ma, Y.-H. Li, Y. Hou, S.-W. Li, X.-Q. Meng, X.-F. Sun, Q.-Y. Sun, and W.-H. Wang Intra-oocyte Localization of MAD2 and Its Relationship with Kinetochores, Microtubules, and Chromosomes in Rat Oocytes During Meiosis Biol Reprod, September 1, 2004; 71(3): 740 - 748. [Abstract] [Full Text] [PDF]

				Y.-H. Li, Y. Hou, W. Ma, J.-X. Yuan, D. Zhang, Q.-Y. Sun, and W.-H. Wang Localization of CD9 in pig oocytes and its effects on sperm-egg interaction Reproduction, February 1, 2004; 127(2): 151 - 157. [Abstract] [Full Text] [PDF]

				Y.-H. Li, W. Ma, M. Li, Y. Hou, L.-H. Jiao, and W.-H. Wang Reduced Polyspermic Penetration in Porcine Oocytes Inseminated in a New In Vitro Fertilization (IVF) System: Straw IVF Biol Reprod, November 1, 2003; 69(5): 1580 - 1585. [Abstract] [Full Text] [PDF]

				K. Miyoshi, S. J. Rzucidlo, S. L. Pratt, and S. L. Stice Utility of Rapidly Matured Oocytes as Recipients for Production of Cloned Embryos from Somatic Cells in the Pig Biol Reprod, August 1, 2002; 67(2): 540 - 545. [Abstract] [Full Text] [PDF]

				A. J. Kouba, L. R. Abeydeera, I. M. Alvarez, B. N. Day, and W. C. Buhi Effects of the Porcine Oviduct-Specific Glycoprotein on Fertilization, Polyspermy, and Embryonic Development In Vitro Biol Reprod, July 1, 2000; 63(1): 242 - 250. [Abstract] [Full Text]

				Y.-M. Han, W.-H. Wang, L. R. Abeydeera, A. L. Petersen, J.-H. Kim, C. Murphy, B. N. Day, and R. S. Prather Pronuclear Location Before the First Cell Division Determines Ploidy of Polyspermic Pig Embryos Biol Reprod, November 1, 1999; 61(5): 1340 - 1346. [Abstract] [Full Text]

				Y.-M. Han, L. R. Abeydeera, J.-H. Kim, H.-B. Moon, R. A. Cabot, B. N. Day, and R. S. Prather Growth Retardation of Inner Cell Mass Cells in Polyspermic Porcine Embryos Produced In Vitro Biol Reprod, May 1, 1999; 60(5): 1110 - 1113. [Abstract] [Full Text]