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a Department of Food Biophysics, Institute of Food Research, Colney, Norwich NR4 7UA, United Kingdom
b The Babraham Institute, Babraham, Cambridge CB2 4AT, United Kingdom
The plasma membrane of mammalian spermatozoa shows pronounced lateral asymmetry with many glycoproteins restricted to specific domains. Some of these antigens are freely diffusing throughout the membrane whereas others appear static in position. It is not clear whether these concepts also apply to membrane lipids. In this investigation we have used fluorescence recovery after photobleaching (FRAP) techniques to spatially resolve lipid dynamics in various surface domains of 5 species of mammalian spermatozoa (bull, boar, ram, mouse, and guinea pig). Sperm plasma membranes were loaded with 5-(N-octadecanoyl)aminofluorescein (ODAF) reporter probe, and its diffusion was measured in various domains by FRAP analysis. Results showed that in live bull, boar, ram, and mouse spermatozoa, diffusion coefficients (D) were significantly higher over the acrosome and postacrosome than on the midpiece and principal piece of the tail. In dead or permeabilized cells, on the other hand, large immobile phases developed, particularly on the sperm tail, that severely reduced D values. ODAF diffusion was also sensitive to temperature and cross-linking of protein components within the membrane with paraformaldehyde. Guinea pig spermatozoa were different in almost all respects from those of the other species tested. It is concluded that lipid diffusion in the plasma membrane of live spermatozoa varies significantly between surface domains, because of either compositional heterogeneity, or differences in bilayer disposition, or the presence of intramembranous barriers that impede free exchange between domains. This study emphasizes the important role of membrane lipids in regulating polarized migration of sperm surface antigens during developmental processes such as maturation and capacitation.
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