Ability of integrins to mediate fertilization, intracellular calcium release, and parthenogenetic development in bovine oocytes.
The ability of arginine-glycine-aspartic acid (RGD; a sequence recognized by integrins) or non-RGD-containing peptides to block fertilization, induce intracellular Ca(2+) oscillations, and initiate parthenogenetic development in bovine oocytes was investigated. Addition of a soluble RGD peptide during fertilization at concentrations ranging from 10 to 1000 microg/ml significantly decreased (P<0.05) fertilization as compared to the in vitro-fertilized controls. The addition of non-RGD peptide had no effect on fertilization. Two intracellular Ca(2+) transients 21.5+/- 1.9 min apart were observed in 56 of 60 oocytes incubated in RGD peptide concentrations ranging from 20 to 1000 microg/ml. No intracellular Ca(2+) transients were observed in medium alone, non-RGD treatment groups or in the RGD peptide at 10 microg/ml. The percentage of oocytes activated with ionomycin and 6-dimethylaminopurine (63% cleavage and 34% blastocyst development) was significantly higher (P<0.05) than those activated with the RGD peptide and 6-dimethylaminopurine (35% cleavage and 19% blastocyst development). These groups were significantly higher (P<0.05) than either peptide alone, 6-dimethylaminopurine alone, or the non-RGD peptide and 6-dimethylaminopurine treatment groups. These data provide evidence that ligation of an integrin on bovine oocytes with a soluble RGD peptide is capable of blocking fertilization, inducing intracellular Ca(2+) transients, and initiating parthenogenetic development.
Campbell
K D
KD
Department of Animal, Dairy and Veterinary Sciences, Center for Developmental and Molecular Biology, Biotechnology Center, Utah State University, Logan, Utah 84322-4815, USA.
Reed
W A
WA
White
K L
KL
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D005306:Fertilization / Q000187:drug effects; D005307:Fertilization in Vitro; D016023:Integrins / Q000502:physiology*; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D009842:Oligopeptides / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010312:Parthenogenesis
D016023:Integrins; D007476:Ionophores; D009842:Oligopeptides; D015759:Ionomycin; C001020:N(6),N(6)-dimethyladenine; C047981:arginyl-glycyl-aspartic acid; D000225:Adenine; D002118:Calcium