Estamos trabajando para incorporar este artículo al repositorio
Consulte la política de Acceso Abierto del editor


Mammalian spermatozoa must undergo an exocytotic event during fertilization, the acrosome reaction (AR). In most species studied this process is induced by specific glycoproteins of the oocyte extracellular matrix, the zona pellucida (ZP), and it involves guanine nucleotide-binding regulatory proteins (G-proteins), resulting in an uptake of extracellular calcium by the sperm. In the bull, this event has been reported to be mediated by voltage-dependent calcium channels (VDCC). Previous observations showed that neoglycoproteins (NGPs) with N-acetylglucosamine or mannose (GlcNAc-BSA or Man-BSA) residues induce the AR in capacitated human spermatozoa. We report here that the pretreatment of spermatozoa with 125 ng/ml pertussis toxin (PTx) inhibited GlcNAc-BSA-or Man-BSA-induced AR, whereas 1 μg/ml cholera toxin had no effect. These data indicate that the transduction mechanism for GlcNAc-BSA- and Man-BSA-induced AR involves G- proteins of the inhibitory type (G(i)). An increase in the AR rate was observed when capacitated spermatozoa were incubated with increasing concentrations of potassium ions (K+) in Biggers-Whitten-Whittingham (BWW) modified medium (2.6 ± 0.3-fold at 80 mM K+). This induction was observed only when the pH was raised to 8.5, and it was inhibited by verapamil, nitrendipine, ω-conotoxin, nickel ions (Ni2+), lanthanum ions (La3+), or cadmium ions (Cd2+) in a concentration-dependent manner, indicating the participation of VDCC activated by membrane depolarization. The GlcNAc-BSA- or Man-BSA-induced AR was completely inhibited by preincubation of spermatozoa with VDCC blockers and calcium antagonists, indicating a link between the binding of sugar residues of the NGPs and channel activation. The AR induced by membrane depolarization with high K+ medium was not inhibited by PTx, suggesting that Ca2+ entry is downstream to G(i)-protein activation. These data show that the induction of the AR in human spermatozoa by GlcNAc- or Man-NGPs involves VDCC and G(i)-like regulatory proteins similar to the induction described for ZP in other mammalian species.


Documento: Artículo
Título:Voltage-dependent calcium channels and G(i) regulatory protein mediate the human sperm acrosomal exocytosis induced by N-acetylglucosaminyl/mannosyl neoglycoproteins
Autor:Brandelli, A.; Miranda, P.V.; Tezón, J.G.
Filiación:Departamento de Bioquimica, Institute de Biociencias, Univ. Federal do Rio Grande do Sul, Porto Alegre, Brazil
Inst. de Biol. y Med. Exp. -CONICET, Buenos Aires, Argentina
Inst. de Biol. y Med. Experimental, Obligado 2490, Buenos Aires 1428, Argentina
Palabras clave:acrosome reaction; Carbohydrate; G-protein; neoglycoproteins; VDCC; calcium channel affecting agent; glycoprotein; guanine nucleotide binding protein; lanthanum; n acetylglucosamine; nickel; nitrendipine; omega conotoxin; verapamil; acrosome reaction; article; drug receptor binding; exocytosis; extracellular calcium; extracellular matrix; fertilization; human; human cell; membrane depolarization; priority journal; signal transduction; spermatozoon capacitation; zona pellucida; Acetylglucosamine; Acrosome; Calcium Channel Blockers; Calcium Channels; Cholera Toxin; Electrophysiology; Exocytosis; Glycoproteins; GTP-Binding Protein alpha Subunits, Gi-Go; Humans; Male; Mannose; Membrane Potentials; Pertussis Toxin; Serum Albumin; Serum Albumin, Bovine; Virulence Factors, Bordetella; Zona Pellucida
Página de inicio:522
Página de fin:529
Título revista:Journal of Andrology
Título revista abreviado:J. ANDROL.
CAS:Acetylglucosamine, 7512-17-6; Calcium Channel Blockers; Calcium Channels; Cholera Toxin, 9012-63-9; Glycoproteins; GTP-Binding Protein alpha Subunits, Gi-Go, EC; Mannose, 31103-86-3; mannose-bovine serum albumin conjugate; N-acetylglucosamine-bovine serum albumin conjugate; Pertussis Toxin, EC; Serum Albumin; Serum Albumin, Bovine; Virulence Factors, Bordetella


  • Aurell Wistrom, C., Meizel, S., Evidence suggesting involvement of a unique human sperm steroid receptor/Cl- channel complex in the progesterone-initiated acrosome reaction (1993) Dev Biol, 159, pp. 679-690
  • Babcock, D.F., Pfeiffer, D.R., Independent elevation of cytosolic [Ca2+] and pH of mammalian sperm by voltage-dependent and pH-sensitive mechanisms (1987) J Biol Chem, 262, pp. 15041-15047
  • Bailey, J.L., Storey, B.T., Calcium influx into mouse spermatozoa activated by solubilized mouse zona pellucida, monitored with the calcium fluorescent indicator, fluo-3. Inhibition of the influx by three inhibitors of the zona pellucida induced acrosome reaction: Tyrphostic A48, pertussis toxin, and 3-quinuclidinyl benzilate (1994) Mol Reprod Dev, 39, pp. 297-308
  • Benoff, S., Cooper, G.W., Hurley, I., Mandel, F.S., Rosenfeld, D.L., Scholl, G.M., Gilbert, B.R., Hershlag, A., The effect of calcium ion channel blockers on sperm fertilization potential (1994) Fertil Steril, 62, pp. 606-617
  • Benoff, S., Cooper, G.W., Hurley, I., Napolitano, B., Rosenfeld, D.L., Scholl, G.M., Hershlag, A., Human sperm fertilizing potential in vitro is correlated with differential expression of a head-specific mannose-ligand receptor (1993) Fertil Steril, 59, pp. 854-862
  • Benoff, S., Hurley, I., Cooper, G.W., Mandel, F.S., Rosenfeld, D.L., Hershlag, A., Head-specific mannose-ligand receptor expression in human spermatozoa is dependent on capacitation-associated membrane cholesterol loss (1993) Hum Reprod, 8, pp. 2141-2154
  • Bentley, J.K., Garbers, D.L., Domino, S.E., Noland, T.D., Van Dop, C., Spermatozoa contain a guanine nucleotide-binding protein ADP-ribosylated by pertussis toxin (1986) Biochem Biophys Res Commun, 138, pp. 728-734
  • Biggers, J.D., Whitten, W.K., Whittingham, D.G., The culture of mouse embryos in vitro (1971) Methods in Mammalian Embryology, pp. 86-116. , Daniel JD, ed. San Francisco: Freeman
  • Blackmore, P.F., Beebe, S.J., Danforth, D.R., Alexander, N., Progesterone and 17α-hydroxyprogesterone: Novel stimulators of calcium influx in human sperm (1990) J Biol Chem, 265, pp. 1376-1380
  • Bleil, J.D., Wassarman, P.M., Sperm-egg interactions in the mouse: Sequence of events and induction of the acrosome reaction by a zone pellucida glycoprotein (1983) Dev Biol, 95, pp. 317-324
  • Bleil, J.D., Wassarman, P.M., Galactose at the nonreducing terminus of O-linked oligosaccharides of mouse egg zone pellucida glycoprotein ZP3 is essential for the glycoprotein's sperm receptor activity (1988) Proc Natl Acad Sci USA, 85, pp. 6778-6782
  • Brandelli, A., Miranda, P.V., Añon Vazquez, M.G., Marin Briggiler, C.I., Sanjurjo, C., Gonzalez-Echeverria, M.F., Blaquier, J.A., Tezón, J.G., A new predictive test for in vitro fertilization based on the induction of sperm acrosome reaction by N-acetylglucosamine neoglycoprotein (1995) Hum Reprod, 10, pp. 1751-1756
  • Brandelli, A., Miranda, P.V., Tezón, J.G., Participation of glycosylated residues in the human sperm acrosome reaction: Possible role of N-acetylglucosaminidase (1994) Biochim Biophys Acta, 1220, pp. 299-304
  • Burkman, L.J., Coddington, C.C., Franken, D.R., Kruger, T.F., Rosenwaks, Z., Hodgen, G.D., The hemizona assay (HZA): Development of a diagnostic test for the binding of human spermatozoa to the human hemizona pellucida to predict fertilization potential (1988) Fertil Steril, 49, pp. 688-697
  • Chen, J.-S., Doncel, G.F., Alvarez, C., Acosta, A.A., Expression of mannose-binding sites on human spermatozoa and their role in sperm-zona pellucida binding (1995) J Androl, 16, pp. 55-63
  • Cox, T., Peterson, R.N., Identification of calcium conducting channels in isolated boar sperm plasma membranes (1989) Biochem Biophys Res Commun, 161, pp. 162-168
  • Cross, N.L., Morales, P., Overstreet, J.W., Hanson, F.W., Two simple methods for detecting acrosome-reacted human sperm (1986) Gamete Res, 15, pp. 213-226
  • Cross, N.L., Morales, P., Overstreet, J.W., Hanson, F.W., Induction of acrosome reactions by the human zona pellucida (1988) Biol Reprod, 38, pp. 235-242
  • Dejonge, C.J., Mack, S.R., Zaneveld, L.J.D., Synchronous assay for human sperm capacitation and the acrosome reaction (1989) J Androl, 10, pp. 232-239
  • Endo, Y., Lee, M.A., Kopf, G.S., Evidence for the role of a guanine nucleotide-binding regulatory protein in the zona pellucida-induced mouse sperm acrosome reaction (1987) Dev Biol, 119, pp. 210-216
  • Endo, Y., Lee, M.A., Kopf, G.S., Characterization of an islet activating protein-sensitive site in mouse sperm that is involved in the zona pellucida-induced acrosome reaction (1988) Dev Biol, 129, pp. 12-24
  • Florman, H.M., Corron, M.E., Kim, T.D.-H., Babcock, D.F., Activation of voltage-dependent calcium channels of mammalian sperm is required for zona pellucida-induced acrosomal exocytosis (1992) Dev Biol, 152, pp. 304-314
  • Florman, M.H., Tombes, R.M., First, N.L., Babcock, D.F., An adhesion-associated agonist from the zona pellucida activates G protein-promoted elevations of internal Ca2+ and pH that mediate mammalian sperm acrosomal exocytosis (1989) Dev Biol, 135, pp. 133-146
  • Florman, H.M., Wassarman, P.M., O-linked oligosacharides of mouse egg ZP3 account for its sperm receptor activity (1985) Cell, 41, pp. 313-324
  • Foresta, C., Rossato, M., DiVirgilio, F., Ion fluxes through the progesterone-activated channel of the sperm plasma membrane (1993) Biochem J, 294, pp. 279-283
  • Hildebrandt, J.D., Codina, J., Tash, J.S., Kirchick, H.J., Lipschultz, L., Sekura, R.D., Birnbaumer, L., The membrane-bound spermatozoal adenylyl cyclcase system does not share coupling characteristics with somatic cell adenylyl cyclases (1985) Endocrinology, 116, pp. 1357-1366
  • Hyne, R.V., Garbers, D.L., Calcium dependent elevation of adenosine 3′,5′-monophosphate and induction of the acrosome reaction in guinea pig spermatozoa (1979) Proc Natl Acad Sci USA, 76, pp. 5699-5703
  • Jamil, K., White, I.G., Induction of acrosomal reaction in sperm with ion-ophore A23187 and calcium (1981) Arch Androl, 7, pp. 283-292
  • Kopf, G.S., Wilde, M.W., Signal transduction processes leading to acrosomal exocytosis in mammalian spermatozoa (1990) Trends Endocrinol Metab, 1, pp. 362-368
  • Kopf, G.S., Woolkalis, M.J., Gerton, G.L., Evidence for a guanine nucleotide-binding regulatory protein in invertebrate and mammalian sperm: Identification by islet-activating protein-catalyzed ADP-ribosylation and immunochemical methods (1986) J Biol Chem, 261, pp. 7327-7331
  • Lee, M.A., Check, J.H., Kopf, G.S., A guanine nucleotide-binding regulatory protein in human sperm mediates acrosomal exocytosis induced by the human zona pellucida (1992) Mol Reprod Dev, 31, pp. 78-86
  • Leyton, L., Saling, P.M., Evidence that aggregation of sperm receptors by ZP3 triggers the acrosome reaction (1989) J Cell Biol, 108, pp. 2163-2168
  • Liu, D.Y., Baker, H.W.G., Tests of human sperm function and fertilization in vitro (1992) Fertil Steril, 58, pp. 465-483
  • Liu, D.Y., Baker, H.W.G., A new test for the assessment of sperm-zona pellucida penetration: Relationship with results of other sperm tests and fertilization in vitro (1994) Hum Reprod, 9, pp. 489-496
  • Liu, D.Y., Baker, H.W.G., Disordered acrosome reaction of spermatozoa bound to the zona pellucida: A newly discovered sperm defect causing infertility with reduced sperm-zona pellucida penetration and reduced fertilization in vitro (1994) Hum Reprod, 9, pp. 1694-1700
  • Llanos, M.N., Evidence in support of a role for Ca2+-activated K+ channels in the hamster sperm acrosome reaction (1994) J Exp Zool, 269, pp. 484-488
  • Macek, M.B., Lopez, L.C., Shur, B.D., Aggregation of β-1,4-galactosyltransferase on mouse sperm induces the acrosome reaction (1991) Dev Biol, 147, pp. 440-444
  • Mendoza, C., Carreras, A., Moos, J., Tesarik, J., Distinction between true acrosome reaction and degenerative acrosome loss by a one-step staining method using Pisum sativum agglutinin (1992) J Reprod Fertil, 95, pp. 755-763
  • Okamura, N., Fukuda, A., Tanba, M., Sugita, Y., Nagai, T., Changes in the nature of calcium transport systems on the porcine sperm plasma membrane during epididymal maturation (1992) Biochim Biophys Acta, 1108, pp. 110-114
  • Okamura, N., Tanba, M., Fukuda, A., Sugida, Y., Nagai, T., Forskolin stimulates porcine sperm capacitation by increasing calcium uptake (1993) FEBS Lett, 316, pp. 283-286
  • Roblero, L.S., Fernandez, E., Guadarrama, A., Zegers-Hochschild, F., Ortiz, M.E., High potassium concentration and the cumulus corona oocyte complex stimulate the fertilizing capacity of human spermatozoa (1990) Fertil Steril, 54, pp. 328-332
  • Roblero, L.S., Guadarrama, A., Ortiz, M.E., Fernandez, E., Zegers-Hochschild, F., High potassium concentration improves the rate of acrosome reaction in human spermatozoa (1988) Fertil Steril, 49, pp. 676-680
  • Saragücta, P., Lanuza, G., Miranda, P.V., Tezón, J.G., Baranao, J.L., Immunoglobulins from human follicular fluid induce the acrosome reaction in human sperm (1994) Mol Reprod Dev, 39, pp. 280-288
  • Siiteri, J.E., Gottlieb, W., Meizel, S., Partial characterization of a fraction from human follicular fluid that initiates the human sperm acrosome reaction in vitro (1988) Gamete Res, 20, pp. 25-43
  • Tesarik, J., Carreras, A., Mendoza, C., Differential sensitivity of progesterone- and zona pellucida-induced acrosome rea ctions to pertussis toxin (1993) Mol Repord Dev, 34, pp. 183-189
  • Turner, K.O., Garcia, M.A., Meizel, S., Progesterone initiation of the human sperm acrosome reaction: The obligatory increase in intracellular calcium is independent of the chloride requirement (1994) Mol Cell Endocrinol, 101, pp. 221-225
  • (1992) WHO Laboratory Manual for the Examination of Human Semen and Sperm Cervical Mucus Interaction. 3rd Ed., , New York: Cambridge University Press
  • Yanagimachi, R., Usui, N., Calcium dependence of the acrosome reaction and activation of guinea pig spermatozoa (1974) Exp Cell Res, 89, pp. 161-174
  • Zeng, Y., Clark, E.N., Florman, H.M., Sperm membrane potential: Hyperpolarization during capacitation regulates zona pellucida-dependent acrosomal secretion (1995) Dev Biol, 171, pp. 554-563


---------- APA ----------
Brandelli, A., Miranda, P.V. & Tezón, J.G. (1996) . Voltage-dependent calcium channels and G(i) regulatory protein mediate the human sperm acrosomal exocytosis induced by N-acetylglucosaminyl/mannosyl neoglycoproteins. Journal of Andrology, 17(5), 522-529.
Recuperado de [ ]
---------- CHICAGO ----------
Brandelli, A., Miranda, P.V., Tezón, J.G. "Voltage-dependent calcium channels and G(i) regulatory protein mediate the human sperm acrosomal exocytosis induced by N-acetylglucosaminyl/mannosyl neoglycoproteins" . Journal of Andrology 17, no. 5 (1996) : 522-529.
Recuperado de [ ]
---------- MLA ----------
Brandelli, A., Miranda, P.V., Tezón, J.G. "Voltage-dependent calcium channels and G(i) regulatory protein mediate the human sperm acrosomal exocytosis induced by N-acetylglucosaminyl/mannosyl neoglycoproteins" . Journal of Andrology, vol. 17, no. 5, 1996, pp. 522-529.
Recuperado de [ ]
---------- VANCOUVER ----------
Brandelli, A., Miranda, P.V., Tezón, J.G. Voltage-dependent calcium channels and G(i) regulatory protein mediate the human sperm acrosomal exocytosis induced by N-acetylglucosaminyl/mannosyl neoglycoproteins. J. ANDROL. 1996;17(5):522-529.
Available from: [ ]