Spontaneous and calcium ionophore-induced acrosome reaction in human sperm: roles of capacitation time and conditions, plasma membrane translocation of phosphatidylserine and protein tyrosine phosphorylation



Document title: Spontaneous and calcium ionophore-induced acrosome reaction in human sperm: roles of capacitation time and conditions, plasma membrane translocation of phosphatidylserine and protein tyrosine phosphorylation
Journal: Jornal brasileiro de reproducao assistida
Database: PERIÓDICA
System number: 000270186
ISSN: 1517-5693
Authors: 1
2



3
Institutions: 1Conceber, Centro de Medicina Reprodutiva, Curitiba, Parana. Brasil
2Eastern Virginia Medical School, The Jones Institute for Reproductive Medicine, Newport News, Virginia. Estados Unidos de América
3University of Stellenbosch, Department of Obstetrics and Gynecology, Stellenbosch. Sudáfrica
Year:
Season: Jul-Sep
Volumen: 11
Number: 3
Pages: 9-16
Country: Brasil
Language: Inglés
Document type: Artículo
Approach: Experimental, aplicado
English abstract The objective of these studies was to assess the relationships between capacitation time and conditions, plasma membrane translocation of phosphatidylserine (PS) and protein tyrosine phosphorylation, and the spontaneous and agonist-induced acrosome reaction of human spermatozoa. Purified populations of highly motile sperm were incubated in human tubal fluid (HTF) supplemented with varying doses of human serum albumin (HSA) and examined at 0, 3, 6, 12 and 24 hours. Acrosome reaction of live sperm was detected by fluorescein isothiocyanate-labelled Pisum Sativum Agglutinin/Hoechst; annexin V binding was used to monitor PS translocation; tyrosine phosphorylation was examined by immunoblotting. The percentage of live, acrosome reacted sperm and the percentage of live sperm depicting PS translocation increased significantly over time, both under spontaneous- and calcium ionophoretreated conditions. Under spontaneous conditions there was a significant positive correlation between the percentage of live, acrosome reacted sperm and the percentage of live sperm with PS translocation. Tyrosine phosphorylation increased over time and with higher doses of HSA, and was associated with increased PS externalization. Taken together, these results suggest that the spontaneous acrosome reaction of human spermatozoa incubated under conditions that support capacitation is time dependent, is positively associated with changes in plasma membrane lipid distribution resulting in externalization of PS, and is accompanied by tyrosine phosphorylation. The responses to the agonistic effect of a calcium ionophore appear to result in different membrane dynamics
Portuguese abstract O objetivo desse estudo foi avaliar a relação entre a reação acrossômica espontânea e induzida dos espermatozóides humanos considerando o tempo e as condições, translocação da fosfatidilserina (FS) e a foforilação da tirosina. Amostras processadas de sêmen com alta motilidade foram incubadas com human tubal fluid (HTF) suplementado com variações de doses de human serum albumine (HSA) e examinas com 0, 3, 6, 12 e 24 horas. A reação acrossômica em espermatozóides vivos foi detectada por fluorescein isothiocyanate-labelled Pisum Sativum Agglutinin/Hoechst; anexina V foi usada para monitorst a translocação da FS; a fosforilação da tirosina foi examinada por immunoblotting. A porcentagem de vivos e a reação acrossômica dos espermatozóides representam o aumento da translocação da FS com maior rapidez, nas duas condições, espontânea e induzida. Em condições espontâneas houve uma positiva relação entre a porcentagem de vivos, a reação acrossômica e a porcentagem de vivos com translocação da PS. O aumento da fosforilação da tirosina com altas doses de HSA, foi associado com o aumento da liberação da FS. Associados, esses resultados sugerem que a reação acrossômica espontânea de espermatozóides humanos incubados em condições que propiciem a capacitação dependente do tempo, é ligada positivamente na distribuição de lipídios na membrana plasmática resultando na liberação da FS, acompanhado pela fosforilação da tirosina. A resposta para o efeito agonista do ionóforo de cálcio aparece nos resultados em diferentes dinâmicas da membrana
Disciplines: Medicina,
Biología
Keyword: Ginecología y obstetricia,
Reproducción y desarrollo,
Reproducción asistida,
Capacitación espermática,
Reacción acrosomal,
Calcio,
Ionóforos
Keyword: Medicine,
Biology,
Gynecology and obstetrics,
Reproduction and development,
Assisted reproduction,
Sperm capacitation,
Acrosome reaction,
Calcium,
Ionophores
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