Capacitation of bovine sperm by heparin: inhibitory effect of glucose and role of intracellular pH.
Bovine sperm incubated with heparin for 7.5-8.5 h underwent an acrosome reaction in the absence but not the presence of glucose (5 mM). When sperm were incubated under capacitating conditions with heparin for 4 h, glucose inhibited sperm penetration of oocytes (p less than 0.01) and lysophosphatidylcholine (LC) induced acrosome reactions. Addition of glucose for the last 0.25 h of a 4.25-h incubation with heparin had no effect on ability of sperm to acrosome-react in response to LC. Nonmetabolizable sugars 3-O-methyl glucose, 2-deoxyglucose, sucrose, and sorbitol did not inhibit capacitation as judged by sperm sensitivity to LC or fertilization (p greater than 0.05), but capacitation was inhibited by the glycolyzable substrates glucose, mannose, and fructose (p less than 0.05). The glycolytic inhibitor, fluoride, reversed glucose inhibition of capacitation in a dose-dependent manner similar to its effect on glucose uptake by sperm. Extracellular pH declined from 7.4 to 7.2 during a 4-h incubation of sperm with heparin and glucose. The decline of extracellular pH during sperm incubation with glucose did not affect capacitation, since only an extracellular pH below 7.02 inhibited capacitation. The intracellular pH (pHi) of sperm increased 0.40 units over a 5-h incubation under capacitating conditions. The change in pHi was inhibited by glucose. Incubation of sperm with heparin and glucose for 12 h resulted in capacitated sperm as judged by both LC sensitivity and fertilizing ability. These studies demonstrate that glycolyzable substrates delay capacitation of bovine sperm and suggest the effect is in delaying an alkalinization of pHi.
Parrish
J J
JJ
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
Susko-Parrish
J L
JL
First
N L
NL
D000818:Animals; D002417:Cattle; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D009584:Nitrogen / Q000494:pharmacology; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D013075:Sperm Capacitation / Q000187:drug effects*; D013094:Spermatozoa / Q000378:metabolism
D011142:Polyvinyl Alcohol; D006493:Heparin; D005947:Glucose; D009584:Nitrogen