Regional heterogeneity of human spermatozoa detected with monoclonal antibodies

Villarroya, S; Scholler, R

doi:10.1530/jrf.0.0760435

Cited by 24 publications

(22 citation statements)

References 30 publications

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“…'This work was supported by grants from DRED (contrat DRED n91-1497) and DRED (EA 975: Pouvoir fecondant des spermatozoides et r6le des lipides). 2 Alexander [11] observed mAb with immobilizing and agglutinating properties, and some antibodies are able to inhibit the binding and penetration of zona-free hamster ova by human spermatozoa in the hamster egg penetration test [12][13][14][15] or even to prevent the fertilization of human oocytes [16]. The present paper describes the characteristics of a mAb (I9G9) generated against human sperm germ cells as well as its influence on sperm-egg interaction.…”

Section: Introductionmentioning

confidence: 97%

“…Sperm antigens have been defined in several species, including humans. A number of antigens on human mature spermatozoa [3][4][5][6][7][8][9][10][11][12][13][14][15][16] have been identified with monoclonal antibodies (mAb) that reportedly recognize antigenically distinct subsets of human spermatozoa. Some have been involved in functional aspects; for example, Isahakia and Accepted July 21, 1994.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a Monoclonal Antibody to a Human Intra-Acrosomal Antigen that Inhibits Fertilization1

Jimenez

Sion

Grizard

et al. 1994

Biology of Reproduction

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Among monoclonal antibodies (mAb) selected after the immunization of mice with human ejaculated spermatozoa, mAb I9G9 (IgG1 kappa) was found by immunoperoxidase staining to label most of the acrosome of human spermatozoa permeabilized with methanol-acetone. The antigen was poorly expressed on the surface of fresh ejaculated sperm, but was detectable on most viable sperm after 5-h incubation in medium containing human serum albumin (HSA) followed by 30-min incubation with the calcium ionophore A23187. This treatment resulted in acrosomal loss. Immunoelectron microscopy labeling with I9G9 mAb localized the antigen within the acrosome. Immunocytochemistry on testis sections showed that antigen was located in the round spermatids within the adluminal compartment of the seminiferous epithelium. Western blotting of sperm extract proteins showed that sperm intra-acrosomal (SIAA) recognized by I9G9 mAb had a polymorphism of immunogenic peptides from 16 to 35 kDa. Most of the antigenic peptides possessed an isoelectric point of approximately 5. When spermatozoa were treated with a series of protease inhibitors, the polymorphism of immunogenic peptides was reduced, suggesting that the multiple form of the antigen was due, at least in part, to proteolytic processing. In the testis, only a single peptide band of 35 kDa was detected with mAb I9G9. Studies of human tissue specificity by Western blotting showed that the epitope recognized by I9G9 mAb was present solely in ejaculated spermatozoa and the testis. I9G9 mAb did not agglutinate or immobilize sperm but inhibited the penetration of zona-free hamster ova by human sperm.

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Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Characterization of a Monoclonal Antibody to a Human Intra-Acrosomal Antigen that Inhibits Fertilization1

Jimenez

Sion

Grizard

et al. 1994

Biology of Reproduction

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“…These are proteins which are accessible to antibodies present in the reproductive tract secretions. Reports of several monoclonal antibodies are available which cross reacted with only the acrosome of living mouse ( 16 ), human (17) and guineapig sperm (18).…”

Section: Discussionmentioning

confidence: 99%

“…Although monoclonal antibodies are specific to a particular molecule they may also recognize epitopes shared by many molecules ( 17). Monoclonal antibodies labelling only one domain were reported to immunoprecipitate proteins of more than one molecular weight from that of other domains (22).…”

Section: Discussionmentioning

confidence: 99%

Monoclonal Antibody Against a Human Sperm Protein Recognizes Multiple Epitopes on Rabbit and Human Sperm and Blocks Sperm Function

Shaha

Taneja²,

Seshadri³

1993

Hybridoma

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A monoclonal antibody was raised against a human sperm protein of apparent molecular size of 40 kDa. Of the 6 hybridoma clones selected for study, one clone (B-12) was chosen for further investigations. The antibody secreted by the clone agglutinated human and rabbit spermatozoa in vitro. The antibody belonged to IgM class. In indirect immunofluorescence studies this antibody reacted to acrosome of living human and rabbit spermatozoa. On fixed sperm of the same species it recognized midpiece and parts of tail along with the acrosome. Mouse, hamster, guinea pig, monkey and rat sperm showed similar localization. Interaction between rabbit sperm and oocyte was inhibited in vitro by this antibody. On western blots of human and rabbit sperm extracts the antibody recognized more than one epitope.

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“…The spermatozoon is one of the cell types in which regional differentiation was demonstrated (for reviews see Bedford & Cooper, 1978;Koehler, 1978Koehler, , 1981Yanagimachi, 1981;Nicolson, 1982;Bellvé & O'Brien, 1983). Using monoclonal antibodies, we have demonstrated that particular antigens are restricted in a minimum of 6 domains on live human spermatozoa (Villarroya & Scholler, 1986a). One of them, HS 1A.1 antigen, localized on the plasma membrane over the acrosome, might be involved in subtle changes occurring during capacitation, and might help to visualize the loss of the acrosomal cap during the acrosome reaction.…”

Section: Introductionmentioning

confidence: 99%

Lateral diffusion of a human sperm-head antigen during incubation in a capacitation medium and induction of the acrosome reaction in vitro

Villarroya

Scholler²

1987

Reproduction

Self Cite

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An integral component of human spermatozoa, a glycoprotein of Mr 143,000 (two subunits of Mr 76,000 and 67,000) was recognized by the a-HS 1A.1 monoclonal antibody. The antigen was localized on the plasma membrane over the sperm head, as demonstrated by transmission electron microscopy. The antigen-antibody binding on gametes during changes in their functional state was followed by an indirect immunofluorescence assay of live human spermatozoa. In freshly ejaculated spermatozoa the antibody binding pattern revealed a patchwork quilt-like topography of the plasma membrane over the acrosome; the percentage of positive cells varied from 20 to 78% with a mean of 50% (n = 82). Incubation in a capacitation medium could increase this percentage up to 98%, revealing new epitopes in an energy-dependent and temperature-independent manner; concomitantly, a part of the antigen migrated in energy-independent and temperature-dependent manner and accumulated in a ring over the postacrosome. When an acrosome reaction was induced in vitro in the presence of Ca2+ with either A23187, ionomycin or human follicular fluid, the HS 1A.1 antigen migrated until immobilization in a well defined pattern around the equatorial segment (single band) or around the equatorial and postacrosomal segments (2 or, seldom, 3 bands). The new antigen localization resulted from a lateral diffusion of pre-existing molecules, occurred in only a few minutes, did not require energy and was temperature-dependent. At the same time, the well outlined large patch burst into a multitude of small spots before vanishing. this veil-like labelling was often observed in spermatozoa kept in the seminal plasma or treated with a metabolic poison. The HS 1A.1 antigen localization reflects surface changes induced by the incubation in a capacitation medium and the acrosome reaction. Apart from the regional heterogeneity of the plasma membrane of a single cell, as noted above, there were differences in the plasma membrane changes in individual spermatozoa from the same ejaculate as well as in semen samples from different donors. The new antibody binding pattern was often alike in successive ejaculates of the same donor. In patients consulting for infertility the percentage of positive cells was often low and migration of the antigen was slight or absent.

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Regional heterogeneity of human spermatozoa detected with monoclonal antibodies

Cited by 24 publications

References 30 publications

Characterization of a Monoclonal Antibody to a Human Intra-Acrosomal Antigen that Inhibits Fertilization1

Characterization of a Monoclonal Antibody to a Human Intra-Acrosomal Antigen that Inhibits Fertilization1

Monoclonal Antibody Against a Human Sperm Protein Recognizes Multiple Epitopes on Rabbit and Human Sperm and Blocks Sperm Function

Lateral diffusion of a human sperm-head antigen during incubation in a capacitation medium and induction of the acrosome reaction in vitro

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