Incorporating lipids into boar sperm decreases chilling sensitivity but not capacitation potential.
Fresh boar sperm were incubated with small unilamellar liposomes composed of either the total lipids extracted from head plasma membranes (HPM) of fresh boar sperm or selected lipids (SL) of five defined phospholipids with specific acyl chains. To optimize fusion, liposomes with 2 mol% octadecyl rhodamine fluorophore in Beltsville Thawing Solution +/- 1 mM CaCl(2) were incubated at 35 degrees C with 1;ts 10(7) or 10(8) spermatozoa/ml and monitored over 60 min, using flow cytometry and fluorescence microscopy. The HPM fused to both sperm concentrations faster than SL but was equivalent by 30 min (10(8) sperm/ml) or 60 min (10(7) sperm/ml; 57.5 +/- 3% and 67.1 +/- 8% sperm fused to HPM and SL, respectively) +/- Ca(2+). Neither HPM nor SL affected onset of capacitation or spontaneous or ionophore-induced acrosome reactions at 0 or 3 h (chlortetracycline and fluorescein isothiocyanate-Pisum sativum agglutinin; n = 3). During cooling and after cryopreservation (n = 4 ejaculates), SL but not HPM significantly improved sperm motility and viability (Sybr14/propidium iodide staining) +/- 20% egg yolk, but egg yolk alone was more effective than SL alone. Liposomes of complex composition can fuse to boar sperm without harming in vitro capacitation or acrosome reaction and reduce sperm chilling sensitivity.
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Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Bailey
J L
JL
Buhr
M M
MM
D020101:Acrosome Reaction / Q000187:drug effects; D000818:Animals; D003080:Cold Temperature; D015925:Cryopreservation; D003451:Cryoprotective Agents; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D007476:Ionophores / Q000494:pharmacology; D050356:Lipid Metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010743:Phospholipids / Q000737:chemistry; D012235:Rhodamines; D013075:Sperm Capacitation / Q000502:physiology*; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine
D003451:Cryoprotective Agents; D005456:Fluorescent Dyes; D007476:Ionophores; D008081:Liposomes; D008563:Membrane Lipids; D010743:Phospholipids; D012235:Rhodamines; C044542:octadecyl Rhodamine B chloride