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Human Reproduction, Vol. 8, No. 12, pp. 2141-2154, 1993
© 1993 European Society of Human Reproduction and Embryology


review-article

Fertilization and early embryology: Head-specific mannose—ligand receptor expression in human spermatozoa is dependent on capacitation-associated membrane cholesterol loss*

Susan Benoff1, Ian Hurley2, George W. Cooper, Francine S. Mandel3, David L. Rosenfeld and Avner Hershlag

Department of Obstetrics and Gynecology, North Shore University Hospital, and Department of Obstetrics and Gynecology, Cornell University Medical College 33 Walt Whitman Road, Huntington Station, NY 11746, USA 2Kemron Environmental Services 33 Walt Whitman Road, Huntington Station, NY 11746 3Department of Research, North Shore University Hospital and Department of Public Health, Cornell University Medical College New York, USA

Correspondence: 1To whom correspondence should be addressed at: Molecular Biology Laboratory, Division of Human Reproduction, Department of Ob/Gyn, North Shore University Hospital, 300 Community Drive, Boas-Marks Biomedical Science Research Center, Room 125, Manhasset, NY 11030, USA

D-Mannose binding lectins appear on the human sperm head following in-vitro incubation under capacitating conditions. Surface expression of lectin is dependent on reduction of the sterol content of sperm membranes. Mannose-specific lectin distribution over the head differs in acrosome-intact and acrosome-reacted spermatozoa. Sugar competition experiments show that mannose is the only saccharide competitive with D-mannosylated albumin binding and that free mannose induces an acrosome reaction in capacitated spermatozoa. A total of 183 men with normozoospermic semen parameters were screened for the ability of their spermatozoa to bind fluorescein isothiocyanate (FITC)-labelled mannosylated albumins, before and after incubation in vitro. The spermatozoa from 176 men ‘responded’ to incubation by exhibiting time-dependent increases in head-directed mannosylated albumin binding, accompanied by increases in the percentage of spermatozoa showing spontaneous acrosome loss. Motile spermatozoa from the remaining seven men failed to express mannose—lectin binding activity after 18 h of incubation and only low percentages of their spermatozoa showed spontaneous acrosome loss. These seven men were classified as ‘non-responders’. The relative amounts of non-esterified cholesterol in the sperm membranes of the responder and non-responder males were analysed by gas—liquid chromatography. Responder spermatozoa showed decreases in free cholesterol content whereas non-responder spermatozoa exhibited either no decrease or increases in relative free cholesterol per cell. Fresh swim-up spermatozoa contain sub-plasma membrane stores of mannose lectins which are revealed by FITC-mannosylated albumin labelling before and after removal of the plasma membrane by vortexing. In contrast, the mannose—lectin binding activity of capacitated spermatozoa is entirely limited to the sperm surface. Western blots of proteins isolated from sperm plasma membranes after capacitation revealed two molecular species reactive with polyclonal antibodies against human macrophage mannose receptors. A model is proposed for the molecular mechanism whereby mannose lectins are transposed from sub-plasma membrane sites to an integral membrane position as a consequence of the loss of cholesterol from the sperm membrane.

Key words: acrosome status/capacitation/human spermatozoa/mannose-specific receptor/neoglycoprotein ligands

*Presented in part at the 39th Annual Meeting of the Society for Gynecologic Investigation, San Antonio, TX, USA, March 18–21, 1992 (Abstracts 155 and 435).


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