Role of zinc during hamster sperm capacitation.
Zinc stabilizes somatic cell membranes and DNA, inhibits respiration, is present in high concentrations in the male reproductive tract, and may stabilize sperm during storage and ejaculation. Zinc removal from sperm may be necessary to prepare sperm for fertilization (capacitation). Incubation with Zn2+ chelators, e.g., D-penicillamine, can capacitate hamster sperm (Andrews and Bavister, Gamete Res 1989; 23:159-70). In the present study, the Zn(2+)-specific fluorochrome N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide (TSQ) and the vital stain propidium iodide were used to assess the zinc content of live hamster sperm with flow cytometry before and after capacitation. Capacitation was monitored with a salt-stored zona pellucida penetration assay or the occurrence of spontaneous or induced (with lysophosphatidylcholine) acrosome reactions. The effect of added zinc on sperm capacitation was also evaluated. Image Analysis was used to determine the subcellular location of zinc (TSQ fluorescence) and atomic absorption to determine whether the total zinc content of sperm changes during capacitation. Sperm incubated under non-capacitating conditions had high TSQ fluorescence and could not penetrate zonae pellucidae. Sperm incubated under capacitating conditions (plus BSA or D-penicillamine) were zinc-depleted (low fluorescence) and penetrated 90% or 78% of zonae, respectively. Image analysis showed a significant reduction in zinc in the acrosomal region during capacitation with BSA, but this did not correlate with the occurrence of spontaneous acrosome reactions. The atomic absorption data showed that the total zinc content of sperm was reduced by 44% or 40% when sperm were incubated under capacitating conditions (BSA or D-penicillamine, respectively). Zona pellucida penetration was completely inhibited when zinc was present throughout the capacitation period but not when it was added at the end of incubation. These data indicate that removal of zinc from hamster sperm is correlated with capacitation and may play a key regulatory role in this process.
Andrews
J C
JC
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park 16802-4504.
Nolan
J P
JP
Hammerstedt
R H
RH
Bavister
B D
BD
D000177:Acrosome / Q000502:physiology; D000818:Animals; D006224:Cricetinae; D005260:Female; D005434:Flow Cytometry; D005453:Fluorescence; D007091:Image Processing, Computer-Assisted; D008297:Male; D010396:Penicillamine / Q000494:pharmacology; D013054:Spectrophotometry, Atomic; D013075:Sperm Capacitation / Q000187:drug effects / Q000502:physiology*; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D015032:Zinc / Q000032:analysis / Q000494:pharmacology / Q000502:physiology*; D015044:Zona Pellucida / Q000502:physiology
D010396:Penicillamine; D015032:Zinc