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Claudia L Trevino - One of the best experts on this subject based on the ideXlab platform.
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Differences and Similarities: The Richness of Comparative Sperm Physiology.
Physiology (Bethesda Md.), 2020Co-Authors: Alberto Darszon, Takuya Nishigaki, Pablo E Visconti, Ignacio López-gonzález, Claudia L TrevinoAbstract:Species preservation depends on the success of fertilization. Sperm are uniquely equipped to fulfill this task, and, although several mechanisms are conserved among species, striking functional dif...
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Sperm Physiology varies according to ultradian and infradian rhythms
Scientific Reports, 2019Co-Authors: Ayelén Moreno-irusta, Esteban M. Domínguez, Arturo Matamoros-volante, Maria N. Gallea, Rosa Molina, Hector A. Guidobaldi, Jackelyn Melissa Kembro, Maria José Figueras, Claudia L Trevino, Ana BabiniAbstract:The Spermatozoon must be physiologically prepared to fertilize the egg, process called capacitation. Human Sperm samples are heterogeneous in their ability to capacitate themselves, which leads to variability between samples from the same or different donors, and even along the seasons. Here we studied Sperm variation in the capacitation state according to the ability of capacitated Spermatozoa to acrosome react upon stimulation (% ARi) and to be recruited by chemotaxis (% Chex). Both indirect indicators of Sperm capacitation increased along the incubation time with fluctuations. Those capacitated Sperm recruited by chemotaxis showed an ultradian rhythm with a cycle every 2 h, which might be influenced by unknown intrinsic Sperm factors. Two infradian rhythms of 12 months for the % ARi and of 6 months for % Chex were observed, which are associated with the joint action of temperature and photoperiod. Thus, to avoid false negative results, human Sperm samples are recommended to be incubated for a long period (e.g. 18 h) preferably in spring time. This innovative point of view would lead to better comprehend human reproductive biology and to think experimental designs in the light of Sperm cyclicity or to improve Sperm aptitude for clinical purposes.
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Sperm Physiology varies according to ultradian and infradian rhythm s
Scientific Reports, 2019Co-Authors: Ayelén Moreno-irusta, Esteban M. Domínguez, Arturo Matamoros-volante, Maria N. Gallea, Rosa Molina, Hector A. Guidobaldi, Jackelyn Melissa Kembro, Maria José Figueras, Claudia L Trevino, Ana BabiniAbstract:The Spermatozoon must be physiologically prepared to fertilize the egg, process called capacitation. Human Sperm samples are heterogeneous in their ability to capacitate themselves, which leads to variability between samples from the same or different donors, and even along the seasons. Here we studied Sperm variation in the capacitation state according to the ability of capacitated Spermatozoa to acrosome react upon stimulation (% ARi) and to be recruited by chemotaxis (% Chex). Both indirect indicators of Sperm capacitation increased along the incubation time with fluctuations. Those capacitated Sperm recruited by chemotaxis showed an ultradian rhythm with a cycle every 2 h, which might be influenced by unknown intrinsic Sperm factors. Two infradian rhythms of 12 months for the % ARi and of 6 months for % Chex were observed, which are associated with the joint action of temperature and photoperiod. Thus, to avoid false negative results, human Sperm samples are recommended to be incubated for a long period (e.g. 18 h) preferably in spring time. This innovative point of view would lead to better comprehend human reproductive biology and to think experimental designs in the light of Sperm cyclicity or to improve Sperm aptitude for clinical purposes.
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semi automatized segmentation method using image based flow cytometry to study Sperm Physiology the case of capacitation induced tyrosine phosphorylation
Molecular Human Reproduction, 2018Co-Authors: Arturo Matamorosvolante, Pablo E Visconti, Laura Cecilia Giojalas, Ayelen Morenoirusta, Paulina Torresrodriguez, Maria Gracia Gervasi, Claudia L TrevinoAbstract:STUDY QUESTION Is image-based flow cytometry a useful tool to study intracellular events in human Sperm such as protein tyrosine phosphorylation or signaling processes? SUMMARY ANSWER Image-based flow cytometry is a powerful tool to study intracellular events in a relevant number of Sperm cells, which enables a robust statistical analysis providing spatial resolution in terms of the specific subcellular localization of the labeling. WHAT IS KNOWN ALREADY Sperm capacitation is required for fertilization. During this process, Spermatozoa undergo numerous physiological changes, via activation of different signaling pathways, which are not completely understood. Classical approaches for studying Sperm Physiology include conventional microscopy, flow cytometry and Western blotting. These techniques present disadvantages for obtaining detailed subcellular information of signaling pathways in a relevant number of cells. This work describes a new semi-automatized analysis using image-based flow cytometry which enables the study, at the subcellular and population levels, of different Sperm parameters associated with signaling. The increase in protein tyrosine phosphorylation during capacitation is presented as an example. STUDY DESIGN SIZE, DURATION Sperm cells were isolated from seminal plasma by the swim-up technique. We evaluated the intensity and distribution of protein tyrosine phosphorylation in Sperm incubated in non-capacitation and capacitation-supporting media for 1 and 18 h under different experimental conditions. We used an antibody against FER kinase and pharmacological inhibitors in an attempt to identify the kinases involved in protein tyrosine phosphorylation during human Sperm capacitation. PARTICIPANTS/MATERIALS, SETTING, METHODS Semen samples from normoSpermic donors were obtained by masturbation after 2-3 days of sexual abstinence. We used the innovative technique image-based flow cytometry and image analysis tools to segment individual images of Spermatozoa. We evaluated and quantified the regions of Sperm where protein tyrosine phosphorylation takes place at the subcellular level in a large number of cells. We also used immunocytochemistry and Western blot analysis. Independent experiments were performed with semen samples from seven different donors. MAIN RESULTS AND THE ROLE OF CHANCE Using image analysis tools, we developed a completely novel semi-automatic strategy useful for segmenting thousands of individual cell images obtained using image-based flow cytometry. Contrary to immunofluorescence which relies on the analysis of a limited Sperm population and also on the observer, image-based flow cytometry allows for unbiased quantification and simultaneous localization of post-translational changes in an extended Sperm population. Interestingly, important data can be independently analyzed by looking to the frame of interest. As an example, we evaluated the capacitation-associated increase in tyrosine phosphorylation in Sperm incubated in non-capacitation and capacitation-supporting media for 1 and 18 h. As previously reported, protein tyrosine phosphorylation increases in a time-depending manner, but our method revealed that this increase occurs differentially among distinct Sperm segments. FER kinase is reported to be the enzyme responsible for the increase in protein tyrosine phosphorylation in mouse Sperm. Our Western blot analysis revealed for the first time the presence of this enzyme in human Sperm. Using our segmentation strategy, we aimed to quantify the effect of pharmacological inhibition of FER kinase and found a marked reduction of protein tyrosine phosphorylation only in the flagellum, which corresponded to the physical localization of FER in human Sperm. Our method provides an alternative strategy to study signaling markers associated with capacitation, such as protein tyrosine phosphorylation, in a fast and quantitative manner. LARGE SCALE DATA None. LIMITATIONS REASONS FOR CAUTION This is an in vitro study performed under controlled conditions. Chemical inhibitors are not completely specific for the intended target; the possibility of side effects cannot be discarded. WIDER IMPLICATIONS OF THE FINDINGS Our results demonstrate that the use of image-based flow cytometry is a very powerful tool to study Sperm Physiology. A large number of cells can be easily analyzed and information at the subcellular level can be obtained. As the segmentation process works with bright-field images, it can be extended to study expression of other proteins of interest using different antibodies or it can be used in living Sperm to study intracellular parameters that can be followed using fluorescent dyes sensitive to the parameter of interest (e.g. pH, Ca2+). Therefore, this a versatile method that can be exploited to study several aspects of Sperm Physiology. STUDY FUNDING AND COMPETING INTEREST(S) This work was supported DGAPA (IN203116 to C. Trevino), Fronteras-CONACyT No. 71 and Eunice Kennedy Shriver National Institute of Child Health and Human Development NIH (RO1 HD38082) to P.E. Visconti and by a Lalor Foundation fellowship to M.G. Gervasi. A. Matamoros is a student of the Maestria en Ciencias Bioquimicas-UNAM program supported by CONACyT (416400) and DGAPA-UNAM. A. Moreno obtained a scholarship from Red MacroUniversidades and L. Giojalas obtained a schloarhip from CONICET and Universidad Nacional de Cordoba. The authors declare there are not conflicts of interest.
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Carbonic anhydrases and their functional differences in human and mouse Sperm Physiology.
Biochemical and biophysical research communications, 2015Co-Authors: Omar José, Paulina Torres-rodríguez, L.s. Forero-quintero, Julio C. Chávez, J. L. De La Vega-beltran, Fabrizio Carta, Claudiu T. Supuran, J.w. Deitmer, Claudia L TrevinoAbstract:Fertilization is a key reproductive event in which Sperm and egg fuse to generate a new individual. Proper regulation of certain parameters (such as intracellular pH) is crucial for this process. Carbonic anhydrases (CAs) are among the molecular entities that control intracellular pH dynamics in most cells. Unfortunately, little is known about the function of CAs in mammalian Sperm Physiology. For this reason, we re-explored the expression of CAI, II, IV and XIII in human and mouse Sperm. We also measured the level of CA activity, determined by mass spectrometry, and found that it is similar in non-capacitated and capacitated mouse Sperm. Importantly, we found that CAII activity accounts for half of the total CA activity in capacitated mouse Sperm. Using the general CA inhibitor ethoxyzolamide, we studied how CAs participate in fundamental Sperm physiological processes such as motility and acrosome reaction in both species. We found that capacitated human Sperm depend strongly on CA activity to support normal motility, while capacitated mouse Sperm do not. Finally, we found that CA inhibition increases the acrosome reaction in capacitated human Sperm, but not in capacitated mouse Sperm.
Ilona I. Concha - One of the best experts on this subject based on the ideXlab platform.
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Presence and Function of Dopamine Transporter (DAT) in Stallion Sperm: Dopamine Modulates Sperm Motility and Acrosomal Integrity
2016Co-Authors: Javier A. Urra, Ra A. Covarrubias, Joan Enric Rodrı́guez-gil, Ilona I. ConchaAbstract:Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and Sperm Physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive Physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar Sperm, suggesting that dopamine acts as a physiological modulator of Sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian Sperm, specifically monoamine transporters as dopamine (DAT), serotonin (SERT) and norepinephrine (NET) transporters in equine Sperm. We also demonstrated for the first time in equine Sperm a functional dopamine transporter using 4-[4-(Dimethylamino)styryl]-N-methylpyridinium iodide (ASP+), as substrate. In addition, we also showed that dopamine (1 mM) treatment in vitro, does not affect Sperm viability but decreases total and progressive Sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909) and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in Sperm Physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine Sperm is significantly reduced at high concentrations of this catecholamine. In summary, ou
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Presence and Function of Dopamine Transporter (DAT) in Stallion Sperm : Dopamine Modulates Sperm Motility and Acrosomal Integrity
PLOS ONE, 2014Co-Authors: Javier A. Urra, Franz Villaroel-espíndola, Alfredo Ramírez-reveco, Joan E. Rodríguez-gil, Alejandra A Covarrubias, Ilona I. ConchaAbstract:Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and Sperm Physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive Physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar Sperm, suggesting that dopamine acts as a physiological modulator of Sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian Sperm, specifically monoamine transporters as dopamine (DAT), serotonin (SERT) and norepinephrine (NET) transporters in equine Sperm. We also demonstrated for the first time in equine Sperm a functional dopamine transporter using 4-[4-(Dimethylamino)styryl]-N-methylpyridinium iodide (ASP+), as substrate. In addition, we also showed that dopamine (1 mM) treatment in vitro, does not affect Sperm viability but decreases total and progressive Sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909) and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in Sperm Physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine Sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine Sperm, and that the dopamine uptake by DAT can regulate Sperm function, specifically acrosomal integrity and Sperm motility.
Christophe Arnoult - One of the best experts on this subject based on the ideXlab platform.
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pantoprazole a proton pump inhibitor impairs human Sperm motility and capacitation in vitro
Journal of Andrology, 2020Co-Authors: Jessica Escoffier, Bastien Arnaud, Mayis Kaba, Jean Pascal Hograindleur, Emilie Le Blevec, Guillaume Martinez, Isabelle Stevant, Christophe ArnoultAbstract:Background The effects of PPIs on human Sperm fertilizing capacity were poorly investigated although these drugs are widely over-used. Two publications retrospectively studied relationships between any PPI intake and Sperm parameters from patients consulting at infertility clinics, but the conclusions of these reports were contradictory. Only two reports investigated the effects of lansoprazole and omeprazole on Sperm motility and found lansoprazole to be deleterious and omeprazole to be neutral for Sperm motility. The inconsistency of the PPI effect in the previous reports emphasizes the need for more basic research on human Spermatozoa, taking into account the hypothesis that the different PPI drugs may have different effects on Sperm Physiology. Objectives Do PPIs, which are among the most widely sold drug in the word, impact negatively human Sperm capacitation and Sperm motility? Materials and methods The effects of PPIs on human Sperm maturation and motility were analyzed by CASA, flow cytometry, and Western blot. Results We tested the impact of 6 different PPIs on human Sperm motility and capacitation. We showed that pantoprazole, but not the other PPIs, decreased Sperm progressive motility and capacitation-induced Sperm hyperactivation. We therefore investigated further the effects of pantoprazole on Sperm capacitation, and we observed that it had a significant deleterious effect on the capacitation-induced hyperpolarization of the membrane potential and capacitation-associated protein phosphorylation. Discussion and conclusion Our results indicate that exposure to pantoprazole has an adverse effect on the physiological competence of human Spermatozoa. As the capacitation process takes place within the female tract, our results suggest that PPIs intake by the female partner may impair in vivo Sperm maturation and possibly fertilization. Moreover, the absence of adverse effect by PPIs on mouse Sperm emphasizes the need to develop reprotox assays using human material to better assess the effects of medication intake on Sperm Physiology.
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Snake venoms as a source of compounds modulating Sperm Physiology: Secreted phospholipases A2 from Oxyuranus scutellatus scutellatus impact Sperm motility, acrosome reaction and in vitro fertilization in mice.
Biochimie, 2010Co-Authors: Jessica Escoffier, Morgane Couvet, Harold De Pomyers, Pierre Ray, Michel Seve, Gérard Lambeau, Michel De Waard, Christophe ArnoultAbstract:The goal of this study was to identify new compounds from venoms able to modulate Sperm Physiology and more particularly Sperm motility. For this purpose, we screened the effects of 16 snake venoms cleared of molecules higher than 15 kDa on Sperm motility. Venoms rich in neurotoxins like those from Oxyuranus scutellatus scutellatus or Daboia russelii, were highly potent inhibitors of Sperm motility. In contrast, venoms rich in myotoxins like those from Echis carinatus, Bothrops alternatus and Macrovipera lebetina, were inactive. From the main pharmacologically-active fraction of the Taipan snake O. scutellatus s., a proteomic approach allowed us to identify 16 different proteins, among which OS1 and OS2, two secreted phospholipases A2 (sPLA(2)). Purified OS1 and OS2 mimicked the inhibitory effect on Sperm motility and were likely responsible for the inhibitory effect of the active fraction. OS1 and OS2 triggered Sperm acrosome reaction and induced lipid rearrangements of the plasma membrane. The catalytic activity of OS2 was required to modulate Sperm Physiology since catalytically inactive mutants had no effect. Finally, Sperm treated with OS2 were less competent than control Sperm to initiate in vitro normal embryo development. This is the first report characterizing sPLA(2) toxins that modulate in vitro Sperm Physiology.
Alberto Darszon - One of the best experts on this subject based on the ideXlab platform.
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Differences and Similarities: The Richness of Comparative Sperm Physiology.
Physiology (Bethesda Md.), 2020Co-Authors: Alberto Darszon, Takuya Nishigaki, Pablo E Visconti, Ignacio López-gonzález, Claudia L TrevinoAbstract:Species preservation depends on the success of fertilization. Sperm are uniquely equipped to fulfill this task, and, although several mechanisms are conserved among species, striking functional dif...
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Intracellular pH in Sperm Physiology.
Biochemical and biophysical research communications, 2014Co-Authors: Takuya Nishigaki, Claudia L Trevino, Omar José, Ana Laura González-cota, Francisco J. Romero, Alberto DarszonAbstract:Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique Sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca2+ channel; Slo3, a K+ channel; the Sperm-specific Na+/H+ exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for Sperm Physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental Sperm functions such as motility, maturation and the acrosome reaction.
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T-type Ca2+ channels in Spermatogenic cells and Sperm.
Pflugers Archiv : European journal of physiology, 2014Co-Authors: Alberto Darszon, Arturo Hernández-cruzAbstract:Cell function is importantly regulated by the intracellular concentration of Ca2+ ([Ca2+]i). Sperm development and function are deeply influenced by [Ca2+]i which is modulated amongst other ion transporters by plasma membrane Ca2+ permeable channels. The presence and role of voltage-dependent Ca2+ channels (CaV) of the T-type (CaV3) in Sperm Physiology have become a matter of debate in recent years. Though they are functionally present in later stages of development in Spermatogenic cells and testicular Sperm and their mRNAs and proteins detected from Spermatogenic cells to mature mammalian Spermatozoa, their currents have not been recorded in mature Spermatozoa. This review critically summarizes the evidence for the involvement of CaV3 channels in Sperm development and function.
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Cl− Channels and Transporters in Sperm Physiology
Sexual Reproduction in Animals and Plants, 2014Co-Authors: Claudia L Trevino, Omar José, J. L. De La Vega-beltran, Gerardo Orta, Dulce Figueiras-fierro, Gonzalo Ferreira, Enrique Balderas, Alberto DarszonAbstract:Spermatozoa must decode environmental and cellular cues to succeed in fertilization, and this process relies heavily on ion channels. New observations bring to light the relevant participation of Cl− channels and anion transporters in some of the main Sperm functions. Here we review the evidence that indicates the participation of Cl− channels in motility, maturation, and the acrosome reaction (AR), and what is known about their molecular identity and regulation. Our better understanding of Sperm anion transport will yield tools to handle some infertility problems, improve animal breeding and preserve biodiversity, and develop selective and secure male contraceptives.
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cl channels and transporters in Sperm Physiology
2014Co-Authors: Claudia L Trevino, Omar José, Gerardo Orta, Gonzalo Ferreira, Enrique Balderas, Dulce Figueirasfierro, J L De La Vegabeltran, Alberto DarszonAbstract:Spermatozoa must decode environmental and cellular cues to succeed in fertilization, and this process relies heavily on ion channels. New observations bring to light the relevant participation of Cl− channels and anion transporters in some of the main Sperm functions. Here we review the evidence that indicates the participation of Cl− channels in motility, maturation, and the acrosome reaction (AR), and what is known about their molecular identity and regulation. Our better understanding of Sperm anion transport will yield tools to handle some infertility problems, improve animal breeding and preserve biodiversity, and develop selective and secure male contraceptives.
Jessica Escoffier - One of the best experts on this subject based on the ideXlab platform.
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pantoprazole a proton pump inhibitor impairs human Sperm motility and capacitation in vitro
Journal of Andrology, 2020Co-Authors: Jessica Escoffier, Bastien Arnaud, Mayis Kaba, Jean Pascal Hograindleur, Emilie Le Blevec, Guillaume Martinez, Isabelle Stevant, Christophe ArnoultAbstract:Background The effects of PPIs on human Sperm fertilizing capacity were poorly investigated although these drugs are widely over-used. Two publications retrospectively studied relationships between any PPI intake and Sperm parameters from patients consulting at infertility clinics, but the conclusions of these reports were contradictory. Only two reports investigated the effects of lansoprazole and omeprazole on Sperm motility and found lansoprazole to be deleterious and omeprazole to be neutral for Sperm motility. The inconsistency of the PPI effect in the previous reports emphasizes the need for more basic research on human Spermatozoa, taking into account the hypothesis that the different PPI drugs may have different effects on Sperm Physiology. Objectives Do PPIs, which are among the most widely sold drug in the word, impact negatively human Sperm capacitation and Sperm motility? Materials and methods The effects of PPIs on human Sperm maturation and motility were analyzed by CASA, flow cytometry, and Western blot. Results We tested the impact of 6 different PPIs on human Sperm motility and capacitation. We showed that pantoprazole, but not the other PPIs, decreased Sperm progressive motility and capacitation-induced Sperm hyperactivation. We therefore investigated further the effects of pantoprazole on Sperm capacitation, and we observed that it had a significant deleterious effect on the capacitation-induced hyperpolarization of the membrane potential and capacitation-associated protein phosphorylation. Discussion and conclusion Our results indicate that exposure to pantoprazole has an adverse effect on the physiological competence of human Spermatozoa. As the capacitation process takes place within the female tract, our results suggest that PPIs intake by the female partner may impair in vivo Sperm maturation and possibly fertilization. Moreover, the absence of adverse effect by PPIs on mouse Sperm emphasizes the need to develop reprotox assays using human material to better assess the effects of medication intake on Sperm Physiology.
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Snake venoms as a source of compounds modulating Sperm Physiology: Secreted phospholipases A2 from Oxyuranus scutellatus scutellatus impact Sperm motility, acrosome reaction and in vitro fertilization in mice.
Biochimie, 2010Co-Authors: Jessica Escoffier, Morgane Couvet, Harold De Pomyers, Pierre Ray, Michel Seve, Gérard Lambeau, Michel De Waard, Christophe ArnoultAbstract:The goal of this study was to identify new compounds from venoms able to modulate Sperm Physiology and more particularly Sperm motility. For this purpose, we screened the effects of 16 snake venoms cleared of molecules higher than 15 kDa on Sperm motility. Venoms rich in neurotoxins like those from Oxyuranus scutellatus scutellatus or Daboia russelii, were highly potent inhibitors of Sperm motility. In contrast, venoms rich in myotoxins like those from Echis carinatus, Bothrops alternatus and Macrovipera lebetina, were inactive. From the main pharmacologically-active fraction of the Taipan snake O. scutellatus s., a proteomic approach allowed us to identify 16 different proteins, among which OS1 and OS2, two secreted phospholipases A2 (sPLA(2)). Purified OS1 and OS2 mimicked the inhibitory effect on Sperm motility and were likely responsible for the inhibitory effect of the active fraction. OS1 and OS2 triggered Sperm acrosome reaction and induced lipid rearrangements of the plasma membrane. The catalytic activity of OS2 was required to modulate Sperm Physiology since catalytically inactive mutants had no effect. Finally, Sperm treated with OS2 were less competent than control Sperm to initiate in vitro normal embryo development. This is the first report characterizing sPLA(2) toxins that modulate in vitro Sperm Physiology.