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Titolo:
HYALURONIDASE ACTIVITY OF MACAQUE SPERM ASSESSED BY AN IN-VITRO CUMULUS PENETRATION ASSAY
Autore:
MEYERS SA; YUDIN AI; CHERR GN; VANDEVOORT CA; MYLES DG; PRIMAKOFF P; OVERSTREET JW;
Indirizzi:
UNIV PENN,SCH VET MED,DEPT CLIN STUDIES,NEW BOLTON CTR,382 W ST RD KENNETT SQ PA 19348 UNIV CALIF DAVIS,CALIF REG PRIMATE RES CTR DAVIS CA 95616 UNIV CALIF DAVIS,SCH MED,DIV OBSTET & GYNECOL DAVIS CA 95616 UNIV CALIF DAVIS,BODEGA MARINE LAB DAVIS CA 95616 UNIV CONNECTICUT,CTR HLTH,DEPT PHYSIOL FARMINGTON CT 00000
Titolo Testata:
Molecular reproduction and development
fascicolo: 3, volume: 46, anno: 1997,
pagine: 392 - 400
SICI:
1040-452X(1997)46:3<392:HAOMSA>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
ZONA-PELLUCIDA; ACROSOME REACTION; MEMBRANE-PROTEIN; GOLDEN-HAMSTER; SPERMATOZOA; INVITRO; PH-20; FERTILIZATION; CYNOMOLGUS; MATRIX;
Keywords:
PH-20; FERTILIZATION; GAMETE INTERACTION; PRIMATE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
32
Recensione:
Indirizzi per estratti:
Citazione:
S.A. Meyers et al., "HYALURONIDASE ACTIVITY OF MACAQUE SPERM ASSESSED BY AN IN-VITRO CUMULUS PENETRATION ASSAY", Molecular reproduction and development, 46(3), 1997, pp. 392-400

Abstract

A model system consisting of cynomolgus macaque sperm and ovulated hamster ova-cumulus complexes (OCCs) was utilized to study the role of the sperm protein PH-20 in cumulus penetration. The hyaluronidase activity of solubilized macaque sperm PH-20 was evaluated using an ELISA-like microplate assay prior to and following the addition of the hyaluronidase inhibitors heparin (0-100 mu g/ml) and apigenin (250 mu M), as well as the Ig fraction of a polyclonal antibody raised against purified recombinant macaque PH-20 (R10; 10-400 mu g/ml). Sperm motility following exposure to enzyme inhibitors was evaluated using computer-aided sperm motility analysis. Macaque sperm were labeled with the permeant fluorescent nuclear dye, Hoechst 33342, and were coincubated with ovulated hamster OCCs for 30 min at 37 degrees C. The addition of heparin, apigenin, or R10 antibody to solubilized sperm extracts resulted ina linear dose-dependent decrease in hyaluronidase activity(P < .01). In the heterologous cumulus penetration assay, fluorescently labeled macaque sperm that were pretreated with heparin (1-100 mu g/ml), apigenin (250 mu M), or R10 antibody (Ig fraction, 10-400 mu g/ml) demonstrated a dose-dependent decrease in the ability to penetrate hamster OCCs(P < 0.01), in the absence of effects on sperm motility. In the homologous assay, experiments using macaque OCCs and fluorescently labeled macaque sperm confirmed that the same concentrations of heparin and R10 antibody similarly suppressed spermatozoal cumulus penetration (P < .01). These results suggest that macaque sperm PH-20-derived hyaluronidase participates in cumulus penetration in this species, and that this model system is useful for further studies into primate gamete interaction. (C) 1997 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/12/20 alle ore 19:48:16