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Abstract:

Sperm protein tyrosine phosphorylation has been associated with capacitation, motility changes, zona binding, and fertilizing ability. We previously demonstrated that gradient-isolated human sperm subpopulations differ in their plasma membrane composition, their ability to phosphorylate proteins in tyrosine residues, and their capacity to undergo hyperactivation. In this study, we have characterized capacitation-associated changes in protein tyrosine phosphorylation and membrane fluidity in spermatozoa of asthenozoospermic and normozoospermic patients consulting for infertility. Semen samples were studied at baseline and after a capacitating incubation with or without the addition of a permeable cAMP analog and a phosphodiesterase inhibitor. Basic sperm and computer-assisted motion parameters, hyperactivation, protein tyrosine phosphorylation (immunofluorescence and Western blot), and membrane fluidity (fluorescent Laurdan probe) were the main study parameters. In comparison with normozoospermic and proven-fertile donor semen, asthenozoospermic samples showed lower motility, velocity, and amplitude of lateral head displacement, both originally and after a 6-h capacitating incubation. Unlike those in normal samples, asthenozoospermic spermatozoa were unable to increase protein tyrosine phosphorylation during capacitation. Such impairment, however, was overcome when they were incubated with a membrane-permeable cAMP analog and a phosphodiesterase inhibitor, indicating a possible membrane defect. Confirming this hypothesis, plasma membranes of asthenozoospermic sperm showed decreased fluidity (increased Laurdan polarization), even after a capacitating incubation. In conclusion, spermatozoa from functional asthenozoospermic samples may owe their poor motility, and their inability to properly capacitate and develop hyperactivation, to an impairment in the tyrosine phosphorylation of critical proteins caused by decreased membrane fluidity. These findings suggest a molecular pathogenetic mechanism for a common seminal pathology associated with male infertility. © 2005 Society for Reproduction and Fertility.

Registro:

Documento: Artículo
Título:Capacitation-associated protein tyrosine phosphorylation and membrane fluidity changes are impaired in the spermotozoa of asthenozzospermic patients
Autor:Buffone, M.G.; Calamera, J.C.; Verstraeten, S.V.; Doncel, G.F.
Filiación:Departamento de Química Biológica, IQUIFIB (CONICET-UBA), Universidad de Buenos Aires, Buenos Aires, Argentina
CONRAD, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, 601 Colley Ave., Norfolk, VA 23507, United States
Laboratorio de Estudios en Reproducción, Buenos Aires, Argentina
Palabras clave:cyclic AMP; laurdan; phosphodiesterase inhibitor; tyrosine; adult; article; asthenospermia; cell activation; cell isolation; cell membrane; cell subpopulation; clinical article; computer analysis; controlled study; disease association; fertilization; fluorescence analysis; human; human cell; hypothesis; immunofluorescence; incubation time; male; male fertility; male infertility; membrane binding; membrane fluidity; membrane permeability; pathogenesis; polarization; priority journal; protein phosphorylation; semen analysis; sperm donor; spermatozoon capacitation; spermatozoon count; spermatozoon motility; velocity; Western blotting; zona pellucida; Adult; Analysis of Variance; Blotting, Western; Bucladesine; Case-Control Studies; Cells, Cultured; Fluorescent Antibody Technique, Indirect; Humans; Image Processing, Computer-Assisted; Male; Membrane Fluidity; Microscopy, Fluorescence; Oligospermia; Pentoxifylline; Phosphodiesterase Inhibitors; Phosphorylation; Phosphotyrosine; Sperm Capacitation; Sperm Motility; Spermatozoa; Tyrosine
Año:2005
Volumen:129
Número:6
Página de inicio:697
Página de fin:705
DOI: http://dx.doi.org/10.1530/rep.1.00584
Título revista:Reproduction
Título revista abreviado:Reproduction
ISSN:14701626
CODEN:RCUKB
CAS:cyclic AMP, 60-92-4; laurdan, 74515-25-6; tyrosine, 16870-43-2, 55520-40-6, 60-18-4; Bucladesine, 362-74-3; Pentoxifylline, 6493-05-6; Phosphodiesterase Inhibitors; Phosphotyrosine, 21820-51-9; Tyrosine, 55520-40-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14701626_v129_n6_p697_Buffone

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Citas:

---------- APA ----------
Buffone, M.G., Calamera, J.C., Verstraeten, S.V. & Doncel, G.F. (2005) . Capacitation-associated protein tyrosine phosphorylation and membrane fluidity changes are impaired in the spermotozoa of asthenozzospermic patients. Reproduction, 129(6), 697-705.
http://dx.doi.org/10.1530/rep.1.00584
---------- CHICAGO ----------
Buffone, M.G., Calamera, J.C., Verstraeten, S.V., Doncel, G.F. "Capacitation-associated protein tyrosine phosphorylation and membrane fluidity changes are impaired in the spermotozoa of asthenozzospermic patients" . Reproduction 129, no. 6 (2005) : 697-705.
http://dx.doi.org/10.1530/rep.1.00584
---------- MLA ----------
Buffone, M.G., Calamera, J.C., Verstraeten, S.V., Doncel, G.F. "Capacitation-associated protein tyrosine phosphorylation and membrane fluidity changes are impaired in the spermotozoa of asthenozzospermic patients" . Reproduction, vol. 129, no. 6, 2005, pp. 697-705.
http://dx.doi.org/10.1530/rep.1.00584
---------- VANCOUVER ----------
Buffone, M.G., Calamera, J.C., Verstraeten, S.V., Doncel, G.F. Capacitation-associated protein tyrosine phosphorylation and membrane fluidity changes are impaired in the spermotozoa of asthenozzospermic patients. Reproduction. 2005;129(6):697-705.
http://dx.doi.org/10.1530/rep.1.00584