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Titolo:
FLOW CYTOMETRIC DETECTION OF BICARBONATE-INDUCED CHANGES IN LECTIN-BINDING IN BOAR AND RAM SPERM POPULATIONS
Autore:
ASHWORTH PJC; HARRISON RAP; MILLER NGA; PLUMMER JM; WATSON PF;
Indirizzi:
AFRC,BABRAHAM INST,DEPT DEV & CELL SIGNALLING CAMBRIDGE CB2 4AT ENGLAND AFRC,BABRAHAM INST,DEPT DEV & CELL SIGNALLING CAMBRIDGE CB2 4AT ENGLAND AFRC,BABRAHAM INST,DEPT IMMUNOL CAMBRIDGE CB2 4AT ENGLAND UNIV LONDON ROYAL VET COLL,DEPT VET BASIC SCI LONDON SW1 ENGLAND
Titolo Testata:
Molecular reproduction and development
fascicolo: 2, volume: 40, anno: 1995,
pagine: 164 - 176
SICI:
1040-452X(1995)40:2<164:FCDOBC>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
ACROSOME REACTION; MOUSE SPERMATOZOA; MAMMALIAN SPERMATOZOA; INVITRO CAPACITATION; SEMINAL PLASMA; SURFACE; FERTILIZATION; CA-2+; HAMSTER; AGGLUTININ;
Keywords:
CAPACITATION; FITC-LECTINS; SPERMATOZOA; CELL SURFACE; GLYCOCONJUGATES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
44
Recensione:
Indirizzi per estratti:
Citazione:
P.J.C. Ashworth et al., "FLOW CYTOMETRIC DETECTION OF BICARBONATE-INDUCED CHANGES IN LECTIN-BINDING IN BOAR AND RAM SPERM POPULATIONS", Molecular reproduction and development, 40(2), 1995, pp. 164-176

Abstract

Boar and ram spermatozoa were incubated in Tyrode's medium in the presence or absence of bicarbonate/CO2, a component believed essential for capacitation. At intervals, samples were stained with a range of FITC-lectins to detect changes in surface glycoconjugates, using a rapid staining technique to avoid problems of lectin toxicity. The samples were then analysed directly by flow cytometry, using propidium iodide to distinguish dead cells. In the presence of bicarbonate, a live subpopulation of spermatozoa developed, which in both animal species showedhigher binding affinities towards Phaseolus Vulgaris Agglutinin (PHA-E), Sophora Japonica Agglutinin (SJA), and Soybean Agglutinin (SEA), and lower binding affinity towards Erythrina Cristagalli Lectin (ECL). In boar samples, the modified subpopulation reached a maximum after 3 hr incubation, whereas in ram samples it maximized after 1.5 hr. No changes were seen when spermatozoa were incubated in bicarbonate-free medium. The bicarbonate-induced changes in lectin binding were not due to the onset of acrosome reactions, because spermatozoa induced to undergo acrosome reactions with the ionophore A23187 displayed very different lectin-binding patterns. Tested on boar spermatozoa, seminal plasma not only inhibited but reversed the bicarbonate-induced development of the modified subpopulation. EGTA also inhibited development of boarsperm subpopulations; excess Ca2+ was unable to overcome this inhibition, suggesting that multivalent metal ions might be involved in bicarbonate's action. We conclude that bicarbonate causes a loss of surfacecoating material with affinity for ECL and an unmasking of binding sites for SEA, SJA and PHA-E. A modified subpopulation of live spermatozoa is thereby established, which appears to maximize at a rate in accord with reported capacitation times. (C) 1995 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/11/20 alle ore 02:20:12