المؤلفون: Maximilian Lyon, Ping Li, Juan J. Ferreira, Roman M. Lazarenko, Sujay V. Kharade, Meghan Kramer, Samantha J. McClenahan, Emily Days, Joshua A. Bauer, Brittany D. Spitznagel, C. David Weaver, Aluet Borrego Alvarez, Lis C. Puga Molina, Pascale Lybaert, Saayli Khambekar, Alicia Liu, Craig W. Lindsley, Jerod Denton, Celia M. Santi

المصدر: Proceedings of the National Academy of Sciences. 120

مصطلحات موضوعية: Multidisciplinary

الوصف: To fertilize an oocyte, the membrane potential of both mouse and human sperm must hyperpolarize (become more negative inside). Determining the molecular mechanisms underlying this hyperpolarization is vital for developing new contraceptive methods and detecting causes of idiopathic male infertility. In mouse sperm, hyperpolarization is caused by activation of the sperm-specific potassium (K + ) channel SLO3 [C. M. Santi et al. , FEBS Lett. 584 , 1041–1046 (2010)]. In human sperm, it has long been unclear whether hyperpolarization depends on SLO3 or the ubiquitous K + channel SLO1 [N. Mannowetz, N. M. Naidoo, S. A. S. Choo, J. F. Smith, P. V. Lishko, Elife 2 , e01009 (2013), C. Brenker et al. , Elife 3 , e01438 (2014), and S. A. Mansell, S. J. Publicover, C. L. R. Barratt, S. M. Wilson, Mol. Hum. Reprod. 20 , 392–408 (2014)]. In this work, we identified the first selective inhibitor for human SLO3—VU0546110—and showed that it completely blocked heterologous SLO3 currents and endogenous K + currents in human sperm. This compound also prevented sperm from hyperpolarizing and undergoing hyperactivated motility and induced acrosome reaction, which are necessary to fertilize an egg. We conclude that SLO3 is the sole K + channel responsible for hyperpolarization and significantly contributes to the fertilizing ability of human sperm. Moreover, SLO3 is a good candidate for contraceptive development, and mutation of this gene is a possible cause of idiopathic male infertility.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::ede63a72f24deecc4d9bbc9e46c1f884
https://doi.org/10.1073/pnas.2212338120

المؤلفون: Pedro Oliveira, Lis C. Puga Molina, Marco G. Alves, David Martin-Hidalgo

المصدر: F&S Reviews. 2:317-329

مصطلحات موضوعية: Infertility, endocrine system, Assisted reproductive technology, urogenital system, business.industry, medicine.medical_treatment, Embryo, Reproductive technology, Bioinformatics, medicine.disease, Sperm, Intracytoplasmic sperm injection, Male infertility, Medicine, business, Live birth, reproductive and urinary physiology

الوصف: The prevalence of obesity and diabetes, two of the most prevalent metabolic disorders (MetDs) in the world, has been rising exponentially over the last two decades. In addition to other comorbidities, MetDs have a detrimental impact on reproductive features, leading to a boost of the use of assisted reproductive technologies (ARTs) to overcome fertility problems. Although ARTs help to improve MetD male reproductive outcomes, data show that the results are less successful compared with those of men without MetD. Currently, intracytoplasmic sperm injection is the election procedure to bypass infertility in men with MetD. Nevertheless, embryos obtained by intracytoplasmic sperm injection using spermatozoa of men with MetD have a lower probability to end in a live birth. This embryo development shutdown has been related to a higher rate of spermatozoa with fragmented DNA and with modifications on pathways that do not allow embryos to go further in the development process. This special detrimental feature of sperm from men with MetD indicates that advanced sperm selection techniques should be used in these patients to avoid sperm with fragmented DNA. Fortunately, sperm selection procedures are under constant development and eventually will allow physicians to select spermatozoa with higher quality and low DNA fragmentation to be used in further ART, increasing the outcome of those procedures. Future research should be performed to enlighten alterations in embryos derived from spermatozoa of men with MetD.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::b76d31277057f9ec7a538cc777871080
https://doi.org/10.1016/j.xfnr.2021.09.002

المؤلفون: Nicholas C. Spies, Celia M. Santi, Joan K. Riley, Paula A. Balestrini, Mariano G. Buffone, Lis C. Puga Molina, Emily S. Jungheim, Stephanie Gunderson

المصدر: Fertility and Sterility. 115:930-939

مصطلحات موضوعية: 0301 basic medicine, 030219 obstetrics & reproductive medicine, In vitro fertilisation, medicine.diagnostic_test, Pronucleus, urogenital system, business.industry, medicine.medical_treatment, Area under the curve, Obstetrics and Gynecology, Semen analysis, Sperm, Intracytoplasmic sperm injection, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Human fertilization, Reproductive Medicine, Capacitation, medicine, business, Algorithm, reproductive and urinary physiology

الوصف: Objective To measure human sperm intracellular pH (pHi) and develop a machine-learning algorithm to predict successful conventional in vitro fertilization (IVF) in normospermic patients. Design Spermatozoa from 76 IVF patients were capacitated in vitro. Flow cytometry was used to measure sperm pHi, and computer-assisted semen analysis was used to measure hyperactivated motility. A gradient-boosted machine-learning algorithm was trained on clinical data and sperm pHi and membrane potential from 58 patients to predict successful conventional IVF, defined as a fertilization ratio (number of fertilized oocytes [2 pronuclei]/number of mature oocytes) greater than 0.66. The algorithm was validated on an independent set of data from 18 patients. Setting Academic medical center. Patient(s) Normospermic men undergoing IVF. Patients were excluded if they used frozen sperm, had known male factor infertility, or used intracytoplasmic sperm injection only. Intervention(s) None. Main Outcome Measure(s) Successful conventional IVF. Result(s) Sperm pHi positively correlated with hyperactivated motility and with conventional IVF ratio (n = 76) but not with intracytoplasmic sperm injection fertilization ratio (n = 38). In receiver operating curve analysis of data from the test set (n = 58), the machine-learning algorithm predicted successful conventional IVF with a mean accuracy of 0.72 (n = 18), a mean area under the curve of 0.81, a mean sensitivity of 0.65, and a mean specificity of 0.80. Conclusion(s) Sperm pHi correlates with conventional fertilization outcomes in normospermic patients undergoing IVF. A machine-learning algorithm can use clinical parameters and markers of capacitation to accurately predict successful fertilization in normospermic men undergoing conventional IVF.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::5dd3b632e3e286310f12ec9727cc8212
https://doi.org/10.1016/j.fertnstert.2020.10.038

المؤلفون: Thomas Rossetti, Lis C. Puga Molina, Makoto Fushimi, Jochen Buck, Dagmar Wachten, Dario Krapf, Lonny R. Levin, Lubna Ghanem, Carla Ritagliati, Melanie Balbach, Navpreet Kaur, Jacob Ferreira, Peter T. Meinke, Jan N. Hansen, Celia M. Santi

المصدر: Mol Hum Reprod

مصطلحات موضوعية: Male, Embryology, endocrine system, Acrosome reaction, Motility, Biology, ADCY10, Mice, Human fertilization, Pregnancy, Capacitation, Genetics, medicine, Animals, Humans, Protein kinase A, education, Molecular Biology, Sperm motility, Cells, Cultured, reproductive and urinary physiology, Original Research, Mice, Knockout, Mice, Inbred ICR, education.field_of_study, urogenital system, Chemistry, Obstetrics and Gynecology, Cell Biology, Soluble adenylyl cyclase, Oocyte, Spermatozoa, Sperm, Cell biology, Mice, Inbred C57BL, medicine.anatomical_structure, Reproductive Medicine, Fertilization, Adenylyl Cyclase Inhibitors, Female, Developmental Biology, Adenylyl Cyclases

الوصف: Soluble adenylyl cyclase (sAC: ADCY10) has been genetically confirmed to be essential for male fertility in mice and humans. In mice, ex vivo studies of dormant, caudal epididymal sperm demonstrated that sAC is required for initiating capacitation and activating motility. We now use an improved sAC inhibitor, TDI-10229, for a comprehensive analysis of sAC function in mouse and human sperm. In contrast to caudal epididymal mouse sperm, human sperm are collected post-ejaculation, after sAC activity has already been stimulated. In addition to preventing the capacitation-induced stimulation of sAC and protein kinase A activities, tyrosine phosphorylation, alkalinization, beat frequency and acrosome reaction in dormant mouse sperm, sAC inhibitors interrupt each of these capacitation-induced changes in ejaculated human sperm. Furthermore, we show for the first time that sAC is required during acrosomal exocytosis in mouse and human sperm. These data define sAC inhibitors as candidates for non-hormonal, on-demand contraceptives suitable for delivery via intravaginal devices in women.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ff990e408dd249230cc26942d0d9a63a
https://doi.org/10.1101/2021.04.27.441671

المؤلفون: Vanina Gabriela Da Ros, Lis C. Puga Molina, Mariano G. Buffone, Sol Yanel Nuñez, Nicolás Gastón Brukman, Patricia S. Cuasnicú, Alberto Darszon

المصدر: Journal of Cellular Physiology. 234:5276-5288

مصطلحات موضوعية: 0301 basic medicine, Physiology, Intracellular pH, Clinical Biochemistry, Motility, CALCIUM, Ciencias Biológicas, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Capacitation, TYROSINE PHOSPHORYLATION, CAPACITATION, Chemistry, Tyrosine phosphorylation, Cell Biology, Sperm, Cell biology, 030104 developmental biology, Cytoplasm, 030220 oncology & carcinogenesis, INTRACELLULAR PH, Signal transduction, Biología Reproductiva, CIENCIAS NATURALES Y EXACTAS, Intracellular, SPERM

الوصف: Capacitation is a mandatory process for the acquisition of mammalian sperm fertilization competence and involves the activation of a complex and still not fully understood system of signaling pathways. Under in vitro conditions, there is an increase in both protein tyrosine phosphorylation (pTyr) and intracellular Ca2+ levels in several species. In human sperm, results from our group revealed that pTyr signaling can be blocked by inhibiting proline-rich tyrosine kinase 2 (PYK2). Based on the role of PYK2 in other cell types, we investigated whether the PYK2-dependent pTyr cascade serves as a sensor for Ca 2+ signaling during human sperm capacitation. Flow cytometry studies showed that exposure of sperm to the PYK2 inhibitor N-[2-[[[2-[(2,3-dihydro-2-oxo-1 H-indol-5-yl)amino]-5-(trifluoromethyl)-4-pyrimidinyl]amino]methyl]phenyl]- N-methyl-methanesulfonamide hydrate (PF431396) produced a significant and concentration-dependent reduction in intracellular Ca 2+ levels during capacitation. Further studies revealed that PF431396-treated sperm exhibited a decrease in the activity of CatSper, a key sperm Ca 2+ channel. In addition, time course studies during capacitation in the presence of PF431396 showed a significant and sustained decrease in both intracellular Ca 2+ and pH levels after 2 hr of incubation, temporarily coincident with the activation of PYK2 during capacitation. Interestingly, decreases in Ca 2+ levels and progressive motility caused by PF431396 were reverted by inducing intracellular alkalinization with NH 4Cl, without affecting the pTyr blockage. Altogether, these observations support pTyr as an intracellular sensor for Ca 2+ entry in human sperm through regulation of cytoplasmic pH. These results contribute to a better understanding of the modulation of the polymodal CatSper and signaling pathways involved in human sperm capacitation. Fil: Brukman, Nicolás Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Nuñez, Sol Yanel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Puga Molina, Lis del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Buffone, Mariano Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Darszon, Alberto. Universidad Nacional Autónoma de México. Instituto de Biotecnología; México Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Da Ros, Vanina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina

وصف الملف: application/pdf

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::556e80410ad8791631a8bf40f7ec0082
https://doi.org/10.1002/jcp.27337

المؤلفون: Chinwendu Amazu, Sarah K. England, Celia M. Santi, Juan J. Ferreira, Xiaofeng Ma, Lis C. Puga-Molina

مصطلحات موضوعية: Coupling (electronics), Membrane potential, medicine.anatomical_structure, Membrane, Contraction (grammar), Chemistry, Cell, medicine, Biophysics, Depolarization, Hyperpolarization (biology), Intracellular

الوصف: Depolarization of the myometrial smooth muscle cell (MSMC) resting membrane potential is necessary for the transition of the uterus from a quiescent state to a contractile state. The molecular mechanisms involved in this transition are not completely understood. Here, we report a novel coupled system between the Na+-activated K+ channel (SLO2.1) and the non-selective Na+ leak channel (NALCN) which determines the MSMC membrane potential. We show that SLO2.1 currents are activated by an inward Na+ leak current carried by the NALCN channel leading to MSMC hyperpolarization. These results show an unanticipated role for the Na+ leak currents in activating a negative feedback system countering the excitable effects of Na+ currents. This is a novel role for the NALCN channel in which Na+ acts as an intracellular signaling molecule. In fact, we report here that the net effect of Na+ entry through NALCN channels is a hyperpolarization of the MSMCs plasma membrane because of the activation of SLO2.1 K channel. Importantly, we also report that a decrease in NALCN/SLO2.1 activity triggers both Ca2+ entries through VDCCs, promoting myometrial contraction. Consistently, with a functional coupling, our data show that NALCN and SLO2.1 are in proximity to one another in human MSMCs. We propose that the spatial arrangement of SLO2.1 and NALCN permits these channels to functionally interact in order to regulate human MSMC membrane potential and cell excitability to modulate uterine contractile activity.

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::6152e4401e122a820db08ec39e24122d
https://doi.org/10.1101/2020.07.31.229138

المؤلفون: Mariano G. Buffone, Lis C. Puga Molina, Pablo E. Visconti, Ana Romarowski, Nicolás A. Pinto, Alberto Vicens Sanchez, Alberto Darszon, Paulina Torres Rodríguez, Claudia L. Treviño

المصدر: Journal of Cellular Physiology. 232:1404-1414

مصطلحات موضوعية: 0301 basic medicine, 03 medical and health sciences, 030104 developmental biology, Physiology, Capacitation, Chemistry, Clinical Biochemistry, Phosphorylation, Cell Biology, Hyperpolarization (biology), Sperm, Molecular biology

الوصف: Fil: Puga Molina, Lis del Carmen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Instituto de Biologia y Medicina Experimental. Fundacion de Instituto de Biologia y Medicina Experimental. Instituto de Biologia y Medicina Experimental; Argentina

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::2d1d9c16de18fbeb449c5c320df791c5
https://doi.org/10.1002/jcp.25634

المؤلفون: Stephanie Gunderson, Celia M. Santi, Emily S. Jungheim, Lis C. Puga Molina, Joan K. Riley

المصدر: Fertility and Sterility. 112:e201-e202

مصطلحات موضوعية: Andrology, Human fertilization, In vitro fertilisation, Reproductive Medicine, Intracellular pH, medicine.medical_treatment, medicine, Obstetrics and Gynecology, Biology, Sperm

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::fd260b505e2af919d22aafa0867147e5
https://doi.org/10.1016/j.fertnstert.2019.07.642

المؤلفون: Lis C. Puga Molina, Nicholas C. Spies, Stephanie Gunderson, Joan K. Riley, Emily S. Jungheim, Celia M. Santi

المصدر: Fertility and Sterility. 114:e297

مصطلحات موضوعية: In vitro fertilisation, Reproductive Medicine, Capacitation, medicine.medical_treatment, medicine, Obstetrics and Gynecology, Computational biology, Biology

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::35cd5cf33d186772d7ca00c05cc713a6
https://doi.org/10.1016/j.fertnstert.2020.08.815

المؤلفون: Mariano G. Buffone, Tomás L. Falzone, Dario Krapf, Ana Romarowski, Florenza Antonella la Spina, Noritaki Hirohashi, Lis C. Puga Molina, Alejandra Mariel Vitale

المصدر: CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET

مصطلحات موضوعية: Male, 0301 basic medicine, medicine.medical_treatment, Acrosome reaction, Oviducts, Mice, 0302 clinical medicine, Human fertilization, FERTILIZATION, Acrosomal exocytosis, Zona pellucida, reproductive and urinary physiology, OVIDUCT, Cumulus Cells, 030219 obstetrics & reproductive medicine, purl.org/becyt/ford/3.1 [https], Oviduct, Bioquímica y Biología Molecular, Spermatozoa, Medicina Básica, medicine.anatomical_structure, purl.org/becyt/ford/3 [https], Female, Acrosome, endocrine system, CIENCIAS MÉDICAS Y DE LA SALUD, Green Fluorescent Proteins, Mice, Transgenic, Biology, Article, Exocytosis, Andrology, 03 medical and health sciences, medicine, Animals, Molecular Biology, Zona Pellucida, Sperm-Ovum Interactions, In vitro fertilisation, urogenital system, Acrosome Reaction, Cell Biology, Sperm, Mice, Inbred C57BL, 030104 developmental biology, Fertilization, ACROSOME REACTION, Immunology, Oocytes, Sperm Capacitation, SPERM, Developmental Biology

الوصف: Recent evidence demonstrated that most fertilizing mouse sperm undergo acrosomal exocytosis (AE) before binding to the zona pellucida of the eggs. However, the sites where fertilizing sperm could initiate AE and what stimuli trigger it remain unknown. Therefore, the aim of this study was to determine physiological sites of AE by using double transgenic mouse sperm, which carried EGFP in the acrosome and DsRed2 fluorescence in mitochondria. Using live imaging of sperm during in vitro fertilization of cumulus-oocyte complexes, it was observed that most sperm did not undergo AE. Thus, the occurrence of AE within the female reproductive tract was evaluated in the physiological context where this process occurs. Most sperm in the lower segments of the oviduct were acrosome-intact; however, a significant number of sperm that reached the upper isthmus had undergone AE. In the ampulla, only 5% of the sperm were acrosome-intact. These results support our previous observations that most of mouse sperm do not initiate AE close to or on the ZP, and further demonstrate that a significant proportion of sperm initiate AE in the upper segments of the oviductal isthmus. Fil: la Spina, Florenza Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Puga Molina, Lis del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Romarowski, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Vitale, Alejandra Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Falzone, Tomas Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; Argentina Fil: Krapf, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina Fil: Hirohashi, Noritaki. Shimane University; Japón Fil: Buffone, Mariano Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina

وصف الملف: application/pdf

URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7775afbc5ba27740069dbec4ae1f1458
https://doi.org/10.1016/j.ydbio.2016.02.006