المؤلفون: Puga Molina LC; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Buenos Aires, Argentina., Pinto NA; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Buenos Aires, Argentina., Torres Rodríguez P; Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, Morelos, México., Romarowski A; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Buenos Aires, Argentina., Vicens Sanchez A; Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, Morelos, México., Visconti PE; Department of Veterinary and Animal Science, Paige Labs, University of Massachusetts, Amherst, Massachusetts., Darszon A; Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, Morelos, México., Treviño CL; Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, Morelos, México., Buffone MG; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Buenos Aires, Argentina.

المصدر: Journal of cellular physiology [J Cell Physiol] 2017 Jun; Vol. 232 (6), pp. 1404-1414. Date of Electronic Publication: 2016 Oct 26.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Wiley-Liss Country of Publication: United States NLM ID: 0050222 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1097-4652 (Electronic) Linking ISSN: 00219541 NLM ISO Abbreviation: J Cell Physiol Subsets: MEDLINE

مواضيع طبية MeSH: Membrane Potentials*/drug effects , Sperm Capacitation*/drug effects, Alkalies/*metabolism , Cyclic AMP-Dependent Protein Kinases/*metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/*metabolism, Acrosome Reaction/drug effects ; Benzoates/metabolism ; Cell Movement/drug effects ; Chlorides/metabolism ; Cyclic AMP/agonists ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors ; Humans ; Hydrogen-Ion Concentration ; Intracellular Space/drug effects ; Intracellular Space/metabolism ; Isoquinolines/pharmacology ; Models, Biological ; Phosphorylation/drug effects ; Protein Kinase Inhibitors/pharmacology ; Signal Transduction/drug effects ; Sulfonamides/pharmacology ; Thiazolidines/metabolism

مستخلص: Mammalian sperm require to spend a limited period of time in the female reproductive tract to become competent to fertilize in a process called capacitation. It is well established that HCO 3 ^- is essential for capacitation because it activates the atypical soluble adenylate cyclase ADCY10 leading to cAMP production, and promotes alkalinization of cytoplasm, and membrane hyperpolarization. However, how HCO 3 ^- is transported into the sperm is not well understood. There is evidence that CFTR activity is involved in the human sperm capacitation but how this channel is integrated in the complex signaling cascades associated with this process remains largely unknown. In the present work, we have analyzed the extent to which CFTR regulates different events in human sperm capacitation. We observed that inhibition of CFTR affects HCO 3 ^- -entrance dependent events resulting in lower PKA activity. CFTR inhibition also affected cAMP/PKA-downstream events such as the increase in tyrosine phosphorylation, hyperactivated motility, and acrosome reaction. In addition, we demonstrated for the first time, that CFTR and PKA activity are essential for the regulation of intracellular pH, and membrane potential in human sperm. Addition of permeable cAMP partially recovered all the PKA-dependent events altered in the presence of inh-172 which is consistent with a role of CFTR upstream of PKA activation. J. Cell. Physiol. 232: 1404-1414, 2017. © 2016 Wiley Periodicals, Inc.
(© 2016 Wiley Periodicals, Inc.)

المؤلفون: Gervasi MG; Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, USA., Visconti PE; Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, USA.

المصدر: Andrology [Andrology] 2017 Mar; Vol. 5 (2), pp. 204-218.

نوع المنشور: Journal Article; Review; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Wiley-Blackwell Country of Publication: England NLM ID: 101585129 Publication Model: Print Cited Medium: Internet ISSN: 2047-2927 (Electronic) Linking ISSN: 20472919 NLM ISO Abbreviation: Andrology Subsets: MEDLINE

مواضيع طبية MeSH: Epididymis/*metabolism , Signal Transduction/*physiology , Sperm Maturation/*physiology , Spermatogenesis/*physiology , Spermatozoa/*metabolism, Animals ; Epididymis/cytology ; Fertilization/physiology ; Humans ; Male ; Spermatozoa/cytology

مستخلص: After leaving the testis, spermatozoa have not yet acquired the ability to move progressively and are unable to fertilize oocytes. To become fertilization competent, they must go through an epididymal maturation process in the male, and capacitation in the female tract. Epididymal maturation can be defined as those changes occurring to spermatozoa in the epididymis that render the spermatozoa the ability to capacitate in the female tract. As part of this process, sperm cells undergo a series of biochemical and physiological changes that require incorporation of new molecules derived from the epididymal epithelium, as well as post-translational modifications of endogenous proteins synthesized during spermiogenesis in the testis. This review will focus on epididymal maturation events, with emphasis in recent advances in the understanding of the molecular basis of this process.
(© 2017 American Society of Andrology and European Academy of Andrology.)

المؤلفون: Gervasi MG; Department of Veterinary and Animal Sciences, ISB, University of Massachusetts, Amherst, Massachusetts., Visconti PE; Department of Veterinary and Animal Sciences, ISB, University of Massachusetts, Amherst, Massachusetts. pvisconti@vasci.umass.edu.

المصدر: Molecular reproduction and development [Mol Reprod Dev] 2016 Oct; Vol. 83 (10), pp. 860-874.

نوع المنشور: Historical Article; Journal Article; Review; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Wiley-Liss Country of Publication: United States NLM ID: 8903333 Publication Model: Print Cited Medium: Internet ISSN: 1098-2795 (Electronic) Linking ISSN: 1040452X NLM ISO Abbreviation: Mol Reprod Dev Subsets: MEDLINE

مواضيع طبية MeSH: Sperm Capacitation*, Fertilization in Vitro/*methods , Fertilization in Vitro/*trends, Animals ; Female ; Fertilization in Vitro/history ; History, 20th Century ; History, 21st Century ; Humans ; Male

مستخلص: Dr. Min Chue Chang's contributions to the field of reproductive biology set the stage for the development of the contraceptive pill and in vitro fertilization. Throughout his publications, Dr. Chang was also able to transmit his view of the fertilization process in ways that organized research for newer generations of reproductive biologists. Particularly relevant for the achievement of in vitro fertilization in mammals was the discovery that the sperm required a period of residence in the female tract to become fertilization-competent; Dr. Chang and Dr. Austin, in Australia, independently reported this process, now known as sperm capacitation. This review discusses Dr. Chang's views on capacitation, and puts them in the context of recent advances in the understanding of the molecular basis of this process. Mol. Reprod. Dev. 83: 860-874, 2016 © 2016 Wiley Periodicals, Inc.
(© 2016 Wiley Periodicals, Inc.)

المؤلفون: Ramos-Espiritu L; Department of Pharmacology, Weill Cornell Medical College, New York, New York, USA.; The High-Throughput Screening and Spectroscopy Resource Center, The Rockefeller University, New York, New York, USA., Kleinboelting S; Department of Biochemistry, University of Bayreuth, Bayreuth, Germany., Navarrete FA; Department of Veterinary and Animal Science, University of Massachusetts, Amherst, Massachusetts, USA., Alvau A; Department of Veterinary and Animal Science, University of Massachusetts, Amherst, Massachusetts, USA., Visconti PE; Department of Veterinary and Animal Science, University of Massachusetts, Amherst, Massachusetts, USA., Valsecchi F; Brain and Mind Research Institute, Weill Cornell Medical College, New York, New York, USA., Starkov A; Brain and Mind Research Institute, Weill Cornell Medical College, New York, New York, USA., Manfredi G; Brain and Mind Research Institute, Weill Cornell Medical College, New York, New York, USA., Buck H; Department of Pharmacology, Weill Cornell Medical College, New York, New York, USA., Adura C; The High-Throughput Screening and Spectroscopy Resource Center, The Rockefeller University, New York, New York, USA., Zippin JH; Department of Dermatology, Weill Cornell Medical College, New York, New York, USA., van den Heuvel J; Helmholtz Zentrum fur Infektionsforschung, Braunschweig, Germany., Glickman JF; The High-Throughput Screening and Spectroscopy Resource Center, The Rockefeller University, New York, New York, USA., Steegborn C; Department of Biochemistry, University of Bayreuth, Bayreuth, Germany., Levin LR; Department of Pharmacology, Weill Cornell Medical College, New York, New York, USA., Buck J; Department of Pharmacology, Weill Cornell Medical College, New York, New York, USA.

المصدر: Nature chemical biology [Nat Chem Biol] 2016 Oct; Vol. 12 (10), pp. 838-44. Date of Electronic Publication: 2016 Aug 22.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Nature Pub. Group Country of Publication: United States NLM ID: 101231976 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1552-4469 (Electronic) Linking ISSN: 15524450 NLM ISO Abbreviation: Nat Chem Biol Subsets: MEDLINE

مواضيع طبية MeSH: Adenylyl Cyclase Inhibitors/*pharmacology , Adenylyl Cyclases/*metabolism , Pyrimidines/*pharmacology , Thiophenes/*pharmacology, Adenylyl Cyclase Inhibitors/chemistry ; Adenylyl Cyclases/chemistry ; Allosteric Regulation/drug effects ; Dose-Response Relationship, Drug ; Humans ; Models, Molecular ; Molecular Structure ; Pyrimidines/chemistry ; Solubility ; Structure-Activity Relationship ; Thiophenes/chemistry

مستخلص: The prototypical second messenger cAMP regulates a wide variety of physiological processes. It can simultaneously mediate diverse functions by acting locally in independently regulated microdomains. In mammalian cells, two types of adenylyl cyclase generate cAMP: G-protein-regulated transmembrane adenylyl cyclases and bicarbonate-, calcium- and ATP-regulated soluble adenylyl cyclase (sAC). Because each type of cyclase regulates distinct microdomains, methods to distinguish between them are needed to understand cAMP signaling. We developed a mass-spectrometry-based adenylyl cyclase assay, which we used to identify a new sAC-specific inhibitor, LRE1. LRE1 bound to the bicarbonate activator binding site and inhibited sAC via a unique allosteric mechanism. LRE1 prevented sAC-dependent processes in cellular and physiological systems, and it will facilitate exploration of the therapeutic potential of sAC inhibition.
Competing Interests: Drs. Buck, Levin and Zippin own equity interest in CEP Biotech which has licensed commercialization of a panel of monoclonal antibodies directed against sAC. All other authors declare that they have no conflicts of interest with the contents of this article.

المؤلفون: Navarrete FA; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Alvau A; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Lee HC; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Levin LR; Department of Pharmacology, Weill Cornell Medical College, New York, NY, USA., Buck J; Department of Pharmacology, Weill Cornell Medical College, New York, NY, USA., Leon PM; Department of Biological Sciences, University of Delaware, Newark, DE, USA., Santi CM; Department of Obstetrics and Gynecology, Basic Sciences Division, Washington University School of Medicine. St. Louis, MO, USA., Krapf D; Instituto de Biología Molecular y Celular de Rosario (CONICET-UNR), 2000 Rosario, Argentina., Mager J; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Fissore RA; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Salicioni AM; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Darszon A; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca, México., Visconti PE; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA.

المصدر: Scientific reports [Sci Rep] 2016 Sep 15; Vol. 6, pp. 33589. Date of Electronic Publication: 2016 Sep 15.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

مواضيع طبية MeSH: Fertilization in Vitro*, Calcium Ionophores/*pharmacology , Infertility, Male/*pathology, Adenylyl Cyclases/genetics ; Adenylyl Cyclases/metabolism ; Animals ; Calcimycin/pharmacology ; Calcium Channels/genetics ; Calcium Channels/metabolism ; Disease Models, Animal ; Fertilization/drug effects ; Large-Conductance Calcium-Activated Potassium Channels/genetics ; Large-Conductance Calcium-Activated Potassium Channels/metabolism ; Male ; Mice, Inbred C57BL ; Mice, Knockout ; Models, Genetic ; Spermatozoa/drug effects

مستخلص: Mammalian sperm acquire fertilizing capacity in the female tract in a process called capacitation. At the molecular level, capacitation requires protein kinase A activation, changes in membrane potential and an increase in intracellular calcium. Inhibition of these pathways results in loss of fertilizing ability in vivo and in vitro. We demonstrated that transient incubation of mouse sperm with Ca(2+) ionophore accelerated capacitation and rescued fertilizing capacity in sperm with inactivated PKA function. We now show that a pulse of Ca(2+) ionophore induces fertilizing capacity in sperm from infertile CatSper1 (Ca(2+) channel), Adcy10 (soluble adenylyl cyclase) and Slo3 (K(+) channel) KO mice. In contrast, sperm from infertile mice lacking the Ca(2+) efflux pump PMACA4 were not rescued. These results indicate that a transient increase in intracellular Ca(2+) can overcome genetic infertility in mice and suggest this approach may prove adaptable to rescue sperm function in certain cases of human male infertility.
Competing Interests: Drs Levin and Buck report owning equity interest in CEP Biotech which has licensed commercialization of a panel of monoclonal antibodies directed against sAC. All other author(s) declare no competing financial interests.

المؤلفون: Alvau A; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA., Battistone MA; Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina., Gervasi MG; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA., Navarrete FA; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA., Xu X; Department of Electrical and Computer Engineering and School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80521, USA., Sánchez-Cárdenas C; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México., De la Vega-Beltran JL; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México., Da Ros VG; Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina., Greer PA; Department of Pathology and Molecular Medicine, Queen's University, Kingston, Ontario, Canada, K7L 3N6., Darszon A; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México., Krapf D; Department of Electrical and Computer Engineering and School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80521, USA., Salicioni AM; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA., Cuasnicu PS; Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina., Visconti PE; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA pvisconti@vasci.umass.edu.

المصدر: Development (Cambridge, England) [Development] 2016 Jul 01; Vol. 143 (13), pp. 2325-33. Date of Electronic Publication: 2016 May 25.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Company Of Biologists Limited Country of Publication: England NLM ID: 8701744 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1477-9129 (Electronic) Linking ISSN: 09501991 NLM ISO Abbreviation: Development Subsets: MEDLINE

مواضيع طبية MeSH: Phosphotyrosine/*metabolism , Protein-Tyrosine Kinases/*metabolism , Sperm Capacitation/*physiology , Spermatozoa/*enzymology, Animals ; Focal Adhesion Kinase 2/metabolism ; Male ; Mice, Inbred C57BL ; Phosphorylation

مستخلص: Sperm capacitation is required for fertilization. At the molecular level, this process is associated with fast activation of protein kinase A. Downstream of this event, capacitating conditions lead to an increase in tyrosine phosphorylation. The identity of the tyrosine kinase(s) mediating this process has not been conclusively demonstrated. Recent experiments using stallion and human sperm have suggested a role for PYK2 based on the use of small molecule inhibitors directed against this kinase. However, crucially, loss-of-function experiments have not been reported. Here, we used both pharmacological inhibitors and genetically modified mice models to investigate the identity of the tyrosine kinase(s) mediating the increase in tyrosine phosphorylation in mouse sperm. Similar to stallion and human, PF431396 blocks the capacitation-associated increase in tyrosine phosphorylation. Yet, sperm from Pyk2(-/-) mice displayed a normal increase in tyrosine phosphorylation, implying that PYK2 is not responsible for this phosphorylation process. Here, we show that PF431396 can also inhibit FER, a tyrosine kinase known to be present in sperm. Sperm from mice targeted with a kinase-inactivating mutation in Fer failed to undergo capacitation-associated increases in tyrosine phosphorylation. Although these mice are fertile, their sperm displayed a reduced ability to fertilize metaphase II-arrested eggs in vitro.
(© 2016. Published by The Company of Biologists Ltd.)

المؤلفون: Stival C; Instituto de Biología Molecular y Celular de Rosario (CONICET-UNR) and Laboratorio de Especialidades Reproductivas (Facultad de Ciencias Bioquímicas y Farmacéuticas (UNR), 2000, Rosario, Argentina., Puga Molina Ldel C; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Paudel B; Department of Veterinary and Animal Sciences, ISB, University of Massachusetts, 01003, Amherst, MA, USA., Buffone MG; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Visconti PE; Department of Veterinary and Animal Sciences, ISB, University of Massachusetts, 01003, Amherst, MA, USA., Krapf D; Instituto de Biología Molecular y Celular de Rosario (CONICET-UNR) and Laboratorio de Especialidades Reproductivas (Facultad de Ciencias Bioquímicas y Farmacéuticas (UNR), 2000, Rosario, Argentina. krapf@ibr-conicet.gov.ar.

المصدر: Advances in anatomy, embryology, and cell biology [Adv Anat Embryol Cell Biol] 2016; Vol. 220, pp. 93-106.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Review

بيانات الدورية: Publisher: Springer Verlag Country of Publication: Germany NLM ID: 0407712 Publication Model: Print Cited Medium: Print ISSN: 0301-5556 (Print) Linking ISSN: 03015556 NLM ISO Abbreviation: Adv Anat Embryol Cell Biol Subsets: MEDLINE

مواضيع طبية MeSH: Acrosome Reaction/*physiology , Cell Membrane/*metabolism , Sperm Capacitation/*physiology , Spermatozoa/*physiology, Animals ; Cell Membrane Permeability ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/genetics ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Gene Expression Regulation ; Ion Transport ; Large-Conductance Calcium-Activated Potassium Channels/genetics ; Large-Conductance Calcium-Activated Potassium Channels/metabolism ; Male ; Mammals ; Phosphorylation ; Signal Transduction ; Spermatozoa/cytology

مستخلص: Physiological changes that endow mammalian sperm with fertilizing capacity are known as sperm capacitation. As part of capacitation, sperm develop an asymmetrical flagellar beating known as hyperactivation and acquire the ability to undergo the acrosome reaction. Together, these processes promote fertilizing competence in sperm. At the molecular level, capacitation involves a series of signal transduction events which include activation of cAMP-dependent phosphorylation pathways, removal of cholesterol, hyperpolarization of the sperm plasma membrane, and changes in ion permeability. In recent years, new technologies have aided in the study of sperm signaling molecules with better resolution, at both spatial and temporal levels, unraveling how different cascades integrate and cooperate to render a fertilizing sperm. Despite this new information, the molecular mechanisms connecting capacitation with acrosomal exocytosis and hyperactivation are not well understood. This review brings together results obtained in mammalian species in the field of sperm capacitation with special focus on those pathways involved in the preparation to undergo the acrosomal reaction.

المؤلفون: Romarowski A; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Battistone MA; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., La Spina FA; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Puga Molina Ldel C; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Luque GM; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Vitale AM; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Cuasnicu PS; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Visconti PE; Department of Veterinary and Animal Science, Paige Labs, University of Massachusets, Amherst, MA 01003, USA., Krapf D; Instituto de Biología Molecular y Celular de Rosario (CONICET-UNR), Rosario 2000 Argentina., Buffone MG; Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. Electronic address: mgbuffone@ibyme.conicet.gov.ar.

المصدر: Developmental biology [Dev Biol] 2015 Sep 15; Vol. 405 (2), pp. 237-49. Date of Electronic Publication: 2015 Jul 10.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Elsevier Country of Publication: United States NLM ID: 0372762 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1095-564X (Electronic) Linking ISSN: 00121606 NLM ISO Abbreviation: Dev Biol Subsets: MEDLINE

مواضيع طبية MeSH: Exocytosis*, Acrosome Reaction/*physiology , Cofilin 1/*metabolism , Cyclic AMP-Dependent Protein Kinases/*metabolism , Lim Kinases/*metabolism , Sperm Capacitation/*physiology, Actins/metabolism ; Animals ; Crosses, Genetic ; Gene Expression Regulation ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Transgenic ; Microscopy, Fluorescence ; Phosphorylation ; Signal Transduction ; Spermatozoa/metabolism

مستخلص: Mammalian sperm must acquire their fertilizing ability after a series of biochemical modifications in the female reproductive tract collectively called capacitation to undergo acrosomal exocytosis, a process that is essential for fertilization. Actin dynamics play a central role in controlling the process of exocytosis in somatic cells as well as in sperm from several mammalian species. In somatic cells, small GTPases of the Rho family are widely known as master regulators of actin dynamics. However, the role of these proteins in sperm has not been studied in detail. In the present work we characterized the participation of small GTPases of the Rho family in the signaling pathway that leads to actin polymerization during mouse sperm capacitation. We observed that most of the proteins of this signaling cascade and their effector proteins are expressed in mouse sperm. The activation of the signaling pathways of cAMP/PKA, RhoA/C and Rac1 is essential for LIMK1 activation by phosphorylation on Threonine 508. Serine 3 of Cofilin is phosphorylated by LIMK1 during capacitation in a transiently manner. Inhibition of LIMK1 by specific inhibitors (BMS-3) resulted in lower levels of actin polymerization during capacitation and a dramatic decrease in the percentage of sperm that undergo acrosomal exocytosis. Thus, we demonstrated for the first time that the master regulators of actin dynamics in somatic cells are present and active in mouse sperm. Combining the results of our present study with other results from the literature, we have proposed a working model regarding how LIMK1 and Cofilin control acrosomal exocytosis in mouse sperm.
(Copyright © 2015 Elsevier Inc. All rights reserved.)

المؤلفون: Navarrete FA; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., García-Vázquez FA; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA.; Department of Physiology, Veterinary School, University of Murcia, Murcia, Spain.; International Excellence Campus for Higher Education and Research (Campus Mare Nostrum) and Institute for Biomedical Research of Murcia, Murcia, Spain., Alvau A; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Escoffier J; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Krapf D; Instituto de Biología Celular y Molecular de Rosario (CONICET), UNR, Buenos Aires, Argentina., Sánchez-Cárdenas C; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca, México., Salicioni AM; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA., Darszon A; Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca, México., Visconti PE; Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst MA, USA.

المصدر: Journal of cellular physiology [J Cell Physiol] 2015 Aug; Vol. 230 (8), pp. 1758-1769.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Wiley-Liss Country of Publication: United States NLM ID: 0050222 Publication Model: Print Cited Medium: Internet ISSN: 1097-4652 (Electronic) Linking ISSN: 00219541 NLM ISO Abbreviation: J Cell Physiol Subsets: MEDLINE

مواضيع طبية MeSH: Calcium/*metabolism , Signal Transduction/*physiology , Sperm Capacitation/*physiology, Animals ; Blotting, Western ; Calcium Channels/metabolism ; Cyclic AMP/metabolism ; Male ; Mice ; Mice, Knockout ; Phosphorylation ; Sperm Motility/physiology ; Tyrosine/metabolism

مستخلص: Mammalian sperm acquire fertilizing ability in the female tract in a process known as capacitation. At the molecular level, capacitation is associated with up-regulation of a cAMP-dependent pathway, changes in intracellular pH, intracellular Ca(2+), and an increase in tyrosine phosphorylation. How these signaling systems interact during capacitation is not well understood. Results presented in this study indicate that Ca(2+) ions have a biphasic role in the regulation of cAMP-dependent signaling. Media without added Ca(2+) salts (nominal zero Ca(2+)) still contain micromolar concentrations of this ion. Sperm incubated in this medium did not undergo PKA activation or the increase in tyrosine phosphorylation suggesting that these phosphorylation pathways require Ca(2+). However, chelation of the extracellular Ca(2+) traces by EGTA induced both cAMP-dependent phosphorylation and the increase in tyrosine phosphorylation. The EGTA effect in nominal zero Ca(2+) media was mimicked by two calmodulin antagonists, W7 and calmidazolium, and by the calcineurin inhibitor cyclosporine A. These results suggest that Ca(2+) ions regulate sperm cAMP and tyrosine phosphorylation pathways in a biphasic manner and that some of its effects are mediated by calmodulin. Interestingly, contrary to wild-type mouse sperm, sperm from CatSper1 KO mice underwent PKA activation and an increase in tyrosine phosphorylation upon incubation in nominal zero Ca(2+) media. Therefore, sperm lacking Catsper Ca(2+) channels behave as wild-type sperm incubated in the presence of EGTA. This latter result suggests that Catsper transports the Ca(2+) involved in the regulation of cAMP-dependent and tyrosine phosphorylation pathways required for sperm capacitation.
(© 2015 Wiley Periodicals, Inc.)

المؤلفون: Stival C; From the Instituto de Biología Molecular y Celular de Rosario, CONICET-UNR, and Laboratorio de Especialidades Reproductivas, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Rosario SF2000, Argentina., La Spina FA; the Instituto de Biología y Medicina Experimental, CONICET, Ciudad Autónoma de Buenos Aires C1428ADN, Argentina., Baró Graf C; From the Instituto de Biología Molecular y Celular de Rosario, CONICET-UNR, and Laboratorio de Especialidades Reproductivas, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Rosario SF2000, Argentina., Arcelay E; the Department of Veterinary and Animal Science, Paige Labs, University of Massachusetts, Amherst, Massachusetts 01003., Arranz SE; From the Instituto de Biología Molecular y Celular de Rosario, CONICET-UNR, and Laboratorio de Especialidades Reproductivas, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Rosario SF2000, Argentina., Ferreira JJ; the Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis Missouri 63110, and., Le Grand S; the Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis Missouri 63110, and., Dzikunu VA; the Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis Missouri 63110, and., Santi CM; the Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis Missouri 63110, and., Visconti PE; the Department of Veterinary and Animal Science, Paige Labs, University of Massachusetts, Amherst, Massachusetts 01003., Buffone MG; the Instituto de Biología y Medicina Experimental, CONICET, Ciudad Autónoma de Buenos Aires C1428ADN, Argentina., Krapf D; From the Instituto de Biología Molecular y Celular de Rosario, CONICET-UNR, and Laboratorio de Especialidades Reproductivas, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Rosario SF2000, Argentina, krapf@ibr-conicet.gov.ar.

المصدر: The Journal of biological chemistry [J Biol Chem] 2015 Jul 24; Vol. 290 (30), pp. 18855-64. Date of Electronic Publication: 2015 Jun 09.

نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't

بيانات الدورية: Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE

مواضيع طبية MeSH: Cyclic AMP-Dependent Protein Kinases/*biosynthesis , Large-Conductance Calcium-Activated Potassium Channels/*genetics , Membrane Potentials/*genetics , src-Family Kinases/*metabolism, Acrosome/metabolism ; Animals ; Cell Membrane/genetics ; Cell Polarity/genetics ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/genetics ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Gene Expression Regulation ; Humans ; Large-Conductance Calcium-Activated Potassium Channels/metabolism ; Male ; Mice ; Signal Transduction/genetics ; Sperm Capacitation/genetics ; Spermatozoa/metabolism ; src-Family Kinases/genetics

مستخلص: Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among the signaling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have shown previously that mice deficient in the tyrosine kinase cSrc are infertile and exhibit improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti-Tyr(P)-416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompanies capacitation. In addition, we show that cSrc inhibition also blocks the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on the heterologously expressed SLO3 channel. Our results indicate that, similar to SLO1 K(+) channels, cSrc blockers significantly decreased SLO3-mediated currents. Together, these results are consistent with findings showing that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.
(© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)