Conserved mechanism of bicarbonate-induced sensitization of CatSper channels in human and mouse sperm

To fertilize an egg, mammalian sperm must undergo capacitation in the female genital tract. A key contributor to capacitation is the calcium (C

Mecanismos moleculares asociados a la entrada de HCO3 - durante la capacitación espermática humana

Mammalian sperm require to spend a limited period of time in the female reproductive tract to become\ncompetent to fertilize an egg in a process called capacitation. This process consists in a series of cellular\n(hyperactivated motility (HA) and acrosomal exocytosis (AE)) and biochemical changes. Among\nbiochemical changes, HCO3\n- influx and the consequent activation of PKA, the increase in intracellular\npH and membrane potential (Em) hyperpolarization are explored in this thesis work. We have analyzed\nhow HCO3\n- is transported into human sperm. We propose that in the seminal plasma there is a\nrapid HCO3\n- influx through NBC cotransporters. As a consequence, this uptake induce an increase in\nthe basal PKA activity. In contrast, the presence of seminal plasma proteins (?PS) modulates the activation\nof PKA. Once in the female tract, there is a removal of ?PS and a sustained HCO3\n- influx mediated\nby CFTR through SLC26. The sustained activation of PKA then modulates changes in pH and\nEm, which are essential for HA and AE.Fil: Puga Molina, Lis del Carmen. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, ArgentinaLos espermatozoides eyaculados de mamíferos no son capaces de fecundar al ovocito a menos que\nresidan un tiempo en el tracto femenino. Este proceso, denominado capacitación, consiste en una\nserie de complejos cambios bioquímicos y celulares (desarrollo de la movilidad hiperactivada (HA) y\nla exocitosis acrosomal (EA). En cuanto a los cambios bioquímicos, la entrada de HCO3\n- y la\nconsecuente activación de PKA, el aumento de pHi y los cambios en el potencial de membrana (Em)\nson algunos de ellos.\nEn este trabajo estudiamos los mecanismos moleculares asociados con la entrada de HCO3\n- en el\nespermatozoide humano. Planteamos que en el plasma seminal existe un ingreso rápido de HCO3\n- a\ntravés del transportador- NBC, produciendo la activación basal de PKA. En contraparte, la presencia\nde proteínas del plasma seminal (?PS) modula la activación de PKA. Una vez en el tracto femenino\nocurre la remoción de las ?PS y la entrada sostenida de HCO3\n- mediada por CFTR a través de los\nSLC26. La consecuente activación sostenida de PKA modula los cambios en el Em y el pHi durante la\ncapacitación, los cuales a su vez son fundamentales para la HA y EA

Identification of hydrogeochemical processes linked to minor elements in Sierra de Gádor (Almería, Spain)

A groundwater sampling has been carried out in Sierra de Gador (SE Spain) with the aim of determining the processes in which minor ions are involved. Previous works have allowed us to differentiate four groups according to the major ion content. This study enables to subdivide one of these groups. The SO42-/Ba2+ relationship shows two distinct trends, controlled by the precipitation of barite in the sulfated samples. Ions as lithium and boron are related to the existence of sulfate and to the groundwater thermalism. Last, the ion Br is a useful indicator of salinity in the samplesSe ha realizado un muestreo de las aguas subterráneas de Sierra de Gádor (SE España) con objeto de ver los procesos en los que están involucrados los iones minoritarios. Trabajos previos nos han permitido reconocer una diferenciación de las aguas en cuatro grupos en función de su contenido en iones mayoritarios. El presente estudio permite subdividir uno de estos grupos. La relación SO42-/Ba2+ muestra dos tendencias bien marcadas, controladas por la precipitación de barita en las muestras más sulfatadas. Iones como litio y boro se relacionan con la presencia de sulfatos y con el termalismo de las aguas. Por último, el ion Br sería un indicador muy útil de la salinidad de las muestra

SLO2.1/NALCN a sodium signaling complex that regulates uterine activity

Depolarization of the myometrial smooth muscle cell (MSMC) resting membrane potential is necessary for the uterus to transition from a quiescent state to a contractile state. The molecular mechanisms involved in this transition are not completely understood. Here, we report that a coupled system between the N

Late Quaternary multi-genetic processes and products on the northern Gulf of Cadiz upper continental slope (SW Iberian Peninsula)

On continental margins, the upper slope to shelf break environment forms a critical region where sediment supply, hydrographic activity and gravitational processes determine how and when sediments are partitioned between the shallow- and deep-marine realm. On the SW Iberian margin, relatively few studies have addressed the dynamics of this region, although it holds key information regarding the link between the sedimentary evolution of the continental shelf and the contourite depositional system on the middle slope. This work therefore presents a high-resolution analysis of the morphological and stratigraphic expressions of late Quaternary (dominantly last glacial and present-day interglacial) sedimentary processes on the upper slope and shelf margin sector between 7° and 7°30’ W. The integration of seismic, bathymetric and hydrographic data reveals the presence of alongslope processes and products (a bottom current-related plastered drift, moat, erosional surface and terrace, an internal wave-/tide-controlled sediment wave field), downslope (gravitational) processes and products (an upper slope – shelf margin valley system, slumps, debrites, gullies), neotectonic elements (diapirs) and fluid flow features (pockmarks, bright spots). The spatial distribution of these features indicates that the study area becomes increasingly alongslope-dominated towards the W, and oppositely, more downslope-dominated towards the E, because sediment supply to the latter area is enhanced under the dominant eastward dispersal of fluvially supplied sediments on the shelf. In addition, glacial-interglacial variations in the amount of sediments supplied to the shelf edge and the intensity of oceanographic processes in the study area also generate a distinct temporal variability, with glacial and interglacial intervals respectively recording principally downslope- and alongslope-controlled morphological elements. Finally, regardless of these overall spatial and temporal patterns, diapirism and fluid flow are inferred to locally destabilize sediments and induce small-scale mass wasting in the study area. These findings are not only relevant to the northern Gulf of Cadiz, but also to the general understanding of sedimentary dynamics and controls in mixed downslope- and alongslope-controlled upper slope to shelf margin settings worldwide.T. Mestdagh is funded through a doctoral scholarship of the Ghent University Special Research Fund (BOF). We would like to thank the captains, crews and scientists involved in research campaigns COMIC 2013 on board of RV Belgica and LASEA 2013 on board of RV Ramón Margalef. Ship time on RV Belgica was provided by BELSPO and RBINS-OD Nature. This research was performed in collaboration with ”The Drifters” Research Group of the Royal Holloway University of London (UK), and is associated to projects CTM 2012-39599-C03 , CGL2016-80445-R ‘SCORE’ ( AEI/FEDER , UE), CTM2016-75129-C3-1-R , CGL2011-30302-C02-02 and CTM2017-88237-P . Editor Michele Rebesco and two anonymous reviewers are kindly acknowledged for providing constructive feedback

A selective inhibitor of the sperm-specific potassium channel SLO3 impairs human sperm function

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 (

Seismic architecture of mud volcano systems in the Ceuta

Analysis of a new dataset of high-resolution seismic and swath bathymetry data acquired during the CONTOURIBER-1 cruise in the Ceuta Contourite Depositional System (Western Alborán Sea), reveals the detailed architecture of several mud volcanoes systems that comprises both surface and buried constructional edifices as well as their subsurface feeder complex which pierces the contourite deposits. Seismic characterization, distribution and interpretation of structural elements associated to the mud volcano system promotes a more comprehensive understanding of the processes involved in their genesis and evolution.El análisis de los datos batimétricos y de sísmica de alta resolución durante la campaña CONTOURIBER-1 en el Sistema Deposicional Contornítico de Ceuta (Mar de Alborán occidental) muestran la arquitectura en detalle de varios sistemas de volcanes de fango que comprenden edificios en el fondo marino y edificios enterrados, así como sus complejos de alimentación a través de los depósitos contorníticos. La caracterización sísmica, distribución e interpretación de estos elementos estructurales que componen el sistema de volcán de fango constituyen una importante ayuda para un mejor y mayor conocimiento de su génesis y evolución.Publicado

Deep Phenotypic Characterisation of CTCs by Combination of Microfluidic Isolation (IsoFlux) and Imaging Flow Cytometry (ImageStream)

Ines Aznar-Peralta holds a "Garantia Juvenil" fellowship (contract number 8040), and M. Carmen Garrido-Navas has a postdoctoral fellowship funded by the Ministry of Economy, Competitiveness, Enterprises and Universities (DOC_01682).The isolation of circulating tumour cells (CTCs) in colorectal cancer (CRC) mostly relies on the expression of epithelial markers such as EpCAM, and phenotypic characterisation is usually performed under fluorescence microscopy with only one or two additional markers. This limits the ability to detect different CTC subpopulations based on multiple markers. The aim of this work was to develop a novel protocol combining two platforms (IsoFluxTM and ImageStream®X) to improve CTC evaluation. Cancer cell lines and peripheral blood from healthy donors were used to evaluate the efficiency of each platform independently and in combination. Peripheral blood was extracted from 16 early CRC patients (before loco-regional surgery) to demonstrate the suitability of the protocol for CTC assessment. Additionally, peripheral blood was extracted from nine patients one month after surgery to validate the utility of our protocol for identifying CTC subpopulation changes over time. Results: Our protocol had a mean recovery efficiency of 69.5% and a limit of detection of at least four cells per millilitre. We developed an analysis method to reduce noise from magnetic beads used for CTC isolation. CTCs were isolated from CRC patients with a median of 37 CTCs (IQ 13.0–85.5) at baseline. CTCs from CRC patients were significantly (p < 0.0001) larger than cytokeratin (CK)-negative cells, and patients were stratified into two groups based on BRAFV600E and PD-L1 expression on CK-positive cells. The changes observed over time included not only the number of CTCs but also their distribution into four different subpopulations defined according to BRAFV600E and PD-L1 positivity. We developed a novel protocol for semi-automatic CTC isolation and phenotypic characterisation by combining two platforms. Assessment of CTCs from early CRC patients using our protocol allowed the identification of two clusters of patients with changing phenotypes over time."Garantia Juvenil" fellowship 8040Ministry of Economy, Competitiveness, Enterprises and Universities DOC_0168

Molecular Basis of Human Sperm Capacitation

In the early 1950s, Austin and Chang independently described the changes that are required for the sperm to fertilize oocytes in vivo. These changes were originally grouped under name of “capacitation” and were the first step in the development of in vitro fertilization (IVF) in humans. Following these initial and fundamental findings, a remarkable number of observations led to characterization of the molecular steps behind this process. The discovery of certain sperm-specific molecules and the possibility to record ion currents through patch-clamp approaches helped to integrate the initial biochemical observation with the activity of ion channels. This is of particular importance in the male gamete due to the fact that sperm are transcriptionally inactive. Therefore, sperm must control all these changes that occur during their transit through the male and female reproductive tracts by complex signaling cascades that include post-translational modifications. This review is focused on the principal molecular mechanisms that govern human sperm capacitation with particular emphasis on comparing all the reported pieces of evidence with the mouse model

CFTR/ENaC-dependent regulation of membrane potential during human sperm capacitation is initiated by bicarbonate uptake through NBC

To fertilize an egg, sperm must reside in the female reproductive tract to undergo several maturational changes that are collectively referred to as capacitation. From a molecular point of view, the HCO3-dependent activation of the atypical soluble adenylyl cyclase (ADCY10) is one of the first events that occurs during capacitation and leads to the subsequent cAMP-dependent activation of protein kinase A (PKA). Capacitation is also accompanied by hyperpolarization of the sperm plasma membrane. We previously reported that PKA activation is necessary for CFTR (cystic fibrosis transmembrane conductance regulator channel) activity and for the modulation of membrane potential (Em). However, the main HCO3 transporters involved in the initial transport and the PKA-dependent Em changes are not well known nor characterized. Here, we analyzed how the activity of CFTR regulates Em during capacitation and examined its relationship with an electrogenic Na/HCO3 cotransporter (NBC) and epithelial Na channels (ENaCs). We observed that inhibition of both CFTR and NBC decreased HCO3 influx, resulting in lower PKA activity, and that events downstream of the cAMP activation of PKA are essential for the regulation of Em. Addition of a permeable cAMP analog partially rescued the inhibitory effects caused by these inhibitors. HCO3 also produced a rapid membrane hyperpolarization mediated by ENaC channels, which contribute to the regulation of Em during capacitation. Altogether, we demonstrate for the first time, that NBC cotransporters and ENaC channels are essential in the CFTR-dependent activation of the cAMP/PKA signaling pathway and Em regulation during human sperm capacitation.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; ArgentinaFil: Pinto, Nicolás Alejandro. 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; ArgentinaFil: Torres, Nicolás I.. 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; ArgentinaFil: González Cota, Ana, L.. University of Washington; Estados UnidosFil: Luque, Guillermina Maria. 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; ArgentinaFil: Balestrini, Paula Ania. 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; ArgentinaFil: 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; ArgentinaFil: 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; ArgentinaFil: Santi, Celia M.. University of Washington; Estados UnidosFil: Treviño, Claudia L.. Universidad Nacional Autónoma de México; MéxicoFil: Darszon, Alberto. Universidad Nacional Autónoma de México; MéxicoFil: 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; Argentin