HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 77/5/848    most recent biolreprod.107.061788v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Busso, D. Articles by Cuasnicu, P. S Search for Related Content PubMed PubMed Citation Articles by Busso, D. Articles by Cuasnicu, P. S Agricola Articles by Busso, D. Articles by Cuasnicu, P. S

A Novel Function for CRISP1 in Rodent Fertilization: Involvement in Sperm-Zona Pellucida Interaction¹

Instituto de Biología y Medicina Experimental (IBYME-CONICET), 1428 Buenos Aires, Argentina

ABSTRACT

Epididymal protein CRISP1 participates in rat and mouse gamete fusion through its interaction with complementary sites on the egg surface. Based on in vivo observations, in the present study we investigated the possibility that CRISP1 plays an additional role in the sperm-zona pellucida (ZP) interaction that precedes gamete fusion. In vitro fertilization experiments using zona-intact rat and mouse eggs indicated that the presence of either an antibody against rat CRISP1 (anti-CRISP1) or rat native CRISP1 (rCRISP1) during gamete co-incubation produced a significant decrease in the percentage of fertilized eggs. However, differently to that expected for a protein involved in gamete fusion, no accumulation of perivitelline sperm was observed, suggesting that the inhibitions occurred at the sperm-ZP interaction level. Bacterially expressed recombinant CRISP1 (recCRISP1) also significantly inhibited egg fertilization. In this case, however, an increase in the number of perivitelline sperm was observed. Subsequent experiments evaluating the effect of anti-CRISP1 or rCRISP1 on the number of sperm bound per egg indicated that the protein is involved in the initial step of sperm-ZP binding. In agreement with these functional studies, indirect immunofluorescence experiments revealed that although rCRISP1 is capable of binding to both the ZP and the oolema, recCRISP1 only binds to the egg surface. The finding that deglycosylated rCRISP1 behaves as the untreated protein, whereas the heat-denatured rCRISP1 associated only with the oolema, indicates that the protein ZP-binding ability resides in the conformation rather than in the glycosydic portion of the molecule. The interaction between rCRISP1 and the ZP reproduces the sperm-ZP-binding behavior, as judged by the failure of the protein to interact with the ZP of fertilized eggs. Together, these results support the idea that CRISP1 participates not only in sperm-egg fusion but also in the prior stage of sperm-ZP interaction.

epididymis, fertilization, gamete biology, ovum, sperm

³These authors contributed equally to this work.

⁴Current address: Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Avda Bernardo O'Higgins 340, 6513492 Santiago, Chile.

¹Supported by the World Health Organization (WHO) LID grant (H9/181/R429) to P.S.C. D.B. was a recipient of a fellowship from the National Research Council of Argentina (CONICET). J.A.M. is a recipient of a fellowship from CONICET. A.D. was a recipient of a fellowship from the Latinomerican Programme for Training and Research in Human Reproduction (PLACIRH).

Correspondence: ²Patricia S. Cuasnicu, Instituto de Biología y Medicina Experimental, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina. FAX: 11 4786 2564; e-mail: cuasnicu{at}dna.uba.ar

This article has been cited by other articles:

				P Sipila, J Jalkanen, I T Huhtaniemi, and M Poutanen Novel epididymal proteins as targets for the development of post-testicular male contraception Reproduction, March 1, 2009; 137(3): 379 - 389. [Abstract] [Full Text] [PDF]

				D. Jamsai, D. M Bianco, S. J Smith, D. J Merriner, J. D Ly-Huynh, A. Herlihy, B. Niranjan, G. M Gibbs, and M. K O'Bryan Characterization of gametogenetin 1 (GGN1) and its potential role in male fertility through the interaction with the ion channel regulator, cysteine-rich secretory protein 2 (CRISP2) in the sperm tail Reproduction, June 1, 2008; 135(6): 751 - 759. [Abstract] [Full Text] [PDF]