Artículo

Maldera, J.A.; Muñoz, M.W.; Chirinos, M.; Busso, D.; Raffo, F.G.E.; Battistone, M.A.; Blaquier, J.A.; Larrea, F.; Cuasnicu, P.S. "Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3" (2014) Molecular Human Reproduction. 20(4):341-349
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Abstract:

Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm- ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm-egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein- protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved not only in sperm-egg fusion but also in the previous stage of sperm-ZP binding through its specific interaction with human ZP3. © The Author 2013. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.

Registro:

Documento: Artículo
Título:Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3
Autor:Maldera, J.A.; Muñoz, M.W.; Chirinos, M.; Busso, D.; Raffo, F.G.E.; Battistone, M.A.; Blaquier, J.A.; Larrea, F.; Cuasnicu, P.S.
Filiación:Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires 1428, Argentina
Departamento de Biologiá de la Reproducción, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán (INCMNSZ), México DF, Mexico
Centro Médico Fertilab, Buenos Aires 1116, Argentina
German Cancer Research Center (DKFZ), Center for Molecular Biology of Heidelberg University (ZMBH) Alliance, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany
Departamento de Nutrición, Diabetes y Metabolismo, Facultad de Medicina, Pontificia Universidad Cató lica de Chile, 8331010 Santiago, Chile
Palabras clave:Fertilization; HCRISP1; Oocyte; Sperm; Zona pellucida; CRISP1 protein; polyclonal antibody; protein; recombinant protein; unclassified drug; zona pellucida protein 2; zona pellucida protein 3; zona pellucida protein 4; acrosome reaction; animal cell; article; artificial insemination; binding site; controlled study; enzyme linked immunosorbent assay; female; fertilization; human; human cell; immunofluorescence; insect cell; male; nonhuman; priority journal; protein expression; protein phosphorylation; protein protein interaction; rat; semen analysis; sperm; spermatozoon capacitation; spermatozoon motility; zona pellucida; fertilization; hCRISP1; oocyte; sperm; zona pellucida; Acrosome Reaction; Adult; Antibodies; Binding Sites; Binding, Competitive; Egg Proteins; Epididymis; Female; Gene Expression Regulation; Humans; Male; Membrane Glycoproteins; Protein Binding; Receptors, Cell Surface; Recombinant Proteins; Signal Transduction; Sperm Capacitation; Spermatozoa; Zona Pellucida
Año:2014
Volumen:20
Número:4
Página de inicio:341
Página de fin:349
DOI: http://dx.doi.org/10.1093/molehr/gat092
Título revista:Molecular Human Reproduction
Título revista abreviado:Mol. Hum. Reprod.
ISSN:13609947
CODEN:MHREF
CAS:protein, 67254-75-5
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13609947_v20_n4_p341_Maldera

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Citas:

---------- APA ----------
Maldera, J.A., Muñoz, M.W., Chirinos, M., Busso, D., Raffo, F.G.E., Battistone, M.A., Blaquier, J.A.,..., Cuasnicu, P.S. (2014) . Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3. Molecular Human Reproduction, 20(4), 341-349.
http://dx.doi.org/10.1093/molehr/gat092
---------- CHICAGO ----------
Maldera, J.A., Muñoz, M.W., Chirinos, M., Busso, D., Raffo, F.G.E., Battistone, M.A., et al. "Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3" . Molecular Human Reproduction 20, no. 4 (2014) : 341-349.
http://dx.doi.org/10.1093/molehr/gat092
---------- MLA ----------
Maldera, J.A., Muñoz, M.W., Chirinos, M., Busso, D., Raffo, F.G.E., Battistone, M.A., et al. "Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3" . Molecular Human Reproduction, vol. 20, no. 4, 2014, pp. 341-349.
http://dx.doi.org/10.1093/molehr/gat092
---------- VANCOUVER ----------
Maldera, J.A., Muñoz, M.W., Chirinos, M., Busso, D., Raffo, F.G.E., Battistone, M.A., et al. Human fertilization: Epididymal hCRISP1 mediates sperm-zona pellucida binding through its interaction with ZP3. Mol. Hum. Reprod. 2014;20(4):341-349.
http://dx.doi.org/10.1093/molehr/gat092