Jíménez Movilla, María

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Jíménez Movilla, María

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Jíménez Movilla, María

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Universidad de Murcia. Departamento de Biología Celular e Histología

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Calreticulin from suboolemmal vesicles affects membrane regulation of polyspermy
(2014-02-05) Saavedra MD; Coy, Pilar; Betancourt, M; González-Marquez, H; Mondéjar Corbalán, Irene; Jíménez Movilla, María; Avilés Sánchez, Manuel; Romar Andrés, Raquel; Fisiología; Facultades de la UMU
This study was designed to determine whether calreticulin (CRT), a chaperone protein, is present in in vitro-matured (IVM) pig oocytes and to study its potential role in the block to polyspermy. Western blot analysis, using an anti-CRT antibody, of oocyte lysate showed an immunoreactive band of w60 kDa. Simultaneous labeling of IVM oocytes with anti-CRT antibody and peanut agglutinin lectin (PNA lectin, a porcine cortical granules (CG)-specific binding lectin) revealed localization of CRT in the subplasmalemmal region with a 27.7% colocalization with PNA staining. After IVF, PNA labeling was not observed and anti-CRT labeling decreased significantly in zygotes and disappeared in two-cell embryos. Western blot analysis of oocyte exudate obtained from zona pellucida (ZP)-free oocytes activated with calcium ionophore confirmed the presence of a band that reacted with an anti-CRT antibody. Anti-CRT antibody and PNA labeling were not observed in activated oocytes despite being detectable in non-activated oocytes. The presence of CRT in vesicles located under the oolemma was demonstrated using immunogold cytochemistry at the ultrastructural level. To study the role of CRT in fertilization, ZP-enclosed and ZP-free oocytes were incubated with exogenous CRT and then inseminated. Whereas ZP-free oocytes showed fewer penetrating sperm and lower polyspermy rates than untreated oocytes, the opposite effect was observed in ZP-enclosed oocytes. In conclusion, CRT is confined to subplasmalemmal vesicles partially overlapping with CG contents. Its exocytosis after the oocyte activation seems to participate in the membrane block to polyspermy in pigs but is not involved in the ZP block.
Open Access
JUNO-Coated Beads as a Functional Assay to Capture and Characterize Fertilization-Competent Human Sperm
(2026) Cots Rodríguez, Paula; Jíménez Movilla, María; Biología Celular e Histología; Facultades de la UMU::Facultad de Medicina
Open Access
Comparative View on the Oviductal Environment during the Periconception Period.
(2020) González-Brusi, Leopoldo; Algarra, Blanca; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Jíménez Movilla, María; Moros Nicolás, Carla; Biología Celular e Histología
The oviduct plays important roles in reproductive events: sperm reservoir formation, final gamete maturation, fertilization and early embryo development. It is well known that the oviductal environment affects gametes and embryos and, ultimately, the health of offspring, so that in vivo embryos are better in terms of morphology, cryotolerance, pregnancy rates or epigenetic profile than those obtained in vitro. The deciphering of embryo–maternal interaction in the oviduct may provide a better understanding of the embryo needs during the periconception period to improve reproductive efficiency. Here, we perform a comparative analysis among species of oviductal gene expression related to embryonic development during its journey through the oviduct, as described to date. Cross-talk communication between the oviduct environment and embryo will be studied by analyses of the secreted or exosomal proteins of the oviduct and the presence of receptors in the membrane of the embryo blastomeres. Finally, we review the data that are available to date on the expression and characterization of the most abundant protein in the oviduct, oviductin (OVGP1), highlighting its fundamental role in fertilization and embryonic development.
Open Access
New Insights into the Mammalian Egg Zona Pellucida
(2021) Chevret, Pascale; Algarra, Blanca; González-Brusi, Leopoldo; Cots Rodríguez, Paula; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Jíménez Movilla, María; Moros Nicolás, Carla; Biología Celular e Histología
Mammalian oocytes are surrounded by an extracellular coat called the zona pellucida (ZP), which, from an evolutionary point of view, is the most ancient of the coats that envelope vertebrate oocytes and conceptuses. This matrix separates the oocyte from cumulus cells and is responsible for species-specific recognition between gametes, preventing polyspermy and protecting the preimplantation embryo. The ZP is a dynamic structure that shows different properties before and after fertilization. Until very recently, mammalian ZP was believed to be composed of only three glycoproteins, ZP1, ZP2 and ZP3, as first described in mouse. However, studies have revealed that this composition is not necessarily applicable to other mammals. Such differences can be explained by an analysis of the molecular evolution of the ZP gene family, during which ZP genes have suffered pseudogenization and duplication events that have resulted in differing models of ZP protein composition. The many discoveries made in recent years related to ZP composition and evolution suggest that a compilation would be useful. Moreover, this review analyses ZP biosynthesis, the role of each ZP protein in different mammalian species and how these proteins may interact among themselves and with other proteins present in the oviductal lumen.
Open Access
Cytochemical and biochemical evidences for a complex tridimensional structure of the hamster zona pellucida
(Murcia : F. Hernández, 2009) Saavedra Leos, María Dolores; Gutiérrez Gallego, Ricardo; Fayrer Hosken, Richard; Ballesta Germán, José; Avilés Sánchez, Manuel; Castells Mora, María Teresa; Izquierdo Rico, María José; Jíménez Movilla, María; Martínez Alonso, Emma; Biología Celular
Zona pellucida (ZP) is an extracellular matrix that surrounds eggs and pre-implantation embryos and is required for in vivo fertility. A key event in successful fertilization is sperm binding to the surface of the ZP. It has been previously described that the hamster sperm binds mainly the outer region of the ZP which corresponds to the porous region in contact with the cumulus cells. Using ultrastructural cytochemistry approaches with an antibody developed against porcine ZP, this study shows that the pig ZP shares epitopes with some rodent species like hamster, rat and mouse. In the hamster, these epitopes are located mainly in the outer region of the ZP of preovulatory and ovulated oocytes. By means of biochemical approaches it was demonstrated that 1) the antibody is specific for the native hamster ZP3, 2) four different bands with a molecular weight of 67, 60, 48 and 38 kDa after Nlinked deglycosylation suggesting that the hamster ZP is formed by four proteins, and 3) the different composition observed in the outer region of the hamster ZP could be due to a specific supramolecular structure that makes some epitopes accessible for the antibodies. In summary, this study provides evidence that the different composition observed in the different regions of the ZP is mediated by a different organization of the components of the ZP produced during the oocyte maturation. This different organization could be responsible for the different sperm binding affinity observed for sperm to the outer region versus the inner region of the ZP.
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Raw quantitative proteomics dataset of the membrane fractions of bovine and mouse oocytes and zygotes.
(2025) Jíménez Movilla, María; Hamze Araujo, Julieta Gabriela; Biología Celular e Histología
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Papel de la zona pelúcida en la fecundación
(2017) Algarra, B.; Ballesta, J.; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Jíménez Movilla, María; Moros Nicolás, Carla; Biología Celular e Histología
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Oocytes use the plasminogen-plasmin system to remove supernumerary spermatozoa
(2012-05-03) Grullón LA; Coy Fuster, Pilar; Jíménez Movilla, María; García Vázquez, Francisco Alberto; Mondéjar Corbalán, Irene; Romar Andrés, Raquel; Romar Andrés, Raquel; Fisiología
background: The role of the plasminogen-plasmin (PLG-PLA) system in fertilization is unknown, although its dysfunction has been associated with subfertility in humans. We have recently detected and quantified plasminogen in the oviductal fluid of two mammals and showed a reduction in sperm penetration during IVF when plasminogen is present. The objective of this study was to describe the mechanism by which PLG-PLA system regulates sperm entry into the oocyte. methods and results: By combining biochemical, functional, electron microscopic, immunocytochemical and live cell imaging methods, we show here that (i) plasminogen is activated into the protease plasmin, by gamete interaction; (ii) urokinase-type and tissuetype plasminogen activators are present in oocytes, but they are not of cortical granule origin; (iii) sperm binding to oocytes triggers the releasing of plasminogen activators and (iv) the generated plasmin causes sperm detachment from the zona pellucida. conclusions: Our results describe a novel mechanism for the success or failure of fertilization in mammals, by which molecules present in the oviductal environment are activated by molecules originating within the gametes. We anticipate that therapeutic up- or down-regulation of this physiological mechanism may be used to help in conception or as a contraceptive tool. Since components of the PLG-PLA system are already available as drugs for heart attacks or cancer therapies, basic research on this novel function would be rapidly transferable for clinical application.
Open Access
Mammalian spermatozoa and cumulus cells bind to a 3D model generated by recombinant zona pellucida protein-coated beads.
(Springer Nature, 2019-11-29) Hamze Araujo, Julieta Gabriela; Canha-Gouveia, Analuce; Algarra, Blanca; Gómez-Torres, María José; Romar Andrés, Raquel; Jíménez Movilla, María; Olivares Sánchez, María Concepción; Bioquímica y Biología Molecular B e Inmunología; Biotecnología (Universidad de Alicante)
The egg is a spherical cell encapsulated by the zona pellucida (ZP) which forms a filamentous matrix composed of several glycoproteins that mediate gamete recognition at fertilization. Studies on molecular mechanisms of sperm-egg binding are limited in many mammalian species by the scarcity of eggs, by ethical concerns in harvesting eggs, and by the high cost of producing genetically modified animals. To address these limitations, we have reproduced a three-dimensional (3D) model mimicking the oocyte’s shape, by means of magnetic sepharose beads coated with recombinant ZP glycoproteins (BZP) and cumulus cells. Three preparations composed of either ZP2 (C and N-termini; BZP2), ZP3 (BZP3) or ZP4 (BZP4) were obtained and characterized by protein SDS-PAGE, immunoblot and imaging with confocal and electron microscopy. The functionality of the model was validated by adhesion of cumulus cells, the ability of the glycoprotein-beads to support spermatozoa binding and induce acrosome exocytosis. Thus, our findings document that ZP-beads provide a novel 3D tool to investigate the role of specific proteins on egg-sperm interactions becoming a relevant tool as a diagnostic predictor of mammalian sperm function once transferred to the industry.
Open Access
Análisis de la composición, estructura y función de las glicoproteínas de la zona pelúcida con especial referencia a la especie humana
(Universidad de Murcia, 2012-02-23) Jíménez Movilla, María; Ballesta Germán, José Francisco; Avilés Sánchez, Manuel; Biología Celular
En la presente tesis doctoral se han estudiados diferentes aspectos relacionados con la composición glucídica, estructura, formación y origen de la zona pelúcida y las glicoproteínas que la forman, poniendo especial interés en la especie humana. Para este propósito hemos desarrollado tres líneas principales de investigación, estudio ultraestructural de la zona pelúcida y gránulos corticales de ovocitos humanos y de la síntesis de las glicoproteínas de la ZP humana y de ratón, análisis bioquímico y biofísico de las glicoproteínas de la zona pelúcida de hámster y análisis bioquímico y fisiológico de ZP2 y ZP3 recombinantes humanas expresadas en células CHO. In the present thesis, several aspects of the carbohydrate content, structure, formation and origin of the ZP, with special attention to the human ZP, have been analyzed. Therefore, the methods and techniques used in the present thesis included the ultrasctuctural study of human ZP, cortical granules and biosynthetic pathway of the human and mouse ZP, biochemical and cytochemical characterization of the hamster zona pellucid and characterization of recombinant human ZP2 y ZP3 expressed in CHO cells.