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Romar Andrés, Raquel

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Romar Andrés, Raquel
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Universidad de Murcia. Departamento de Fisiología
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Subject: IVF ×

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    Fertilization outcome could be regulated by binding of oviductal plasminogen to oocytes and by releasing of plasminogen activators during interplay between gametes
    (2012-02-02) Grullón, LA; Mondéjar Corbalán, Irene; García Vázquez, Francisco Alberto; Romar Andrés, Raquel; Coy Fuster, Pilar; Coy Fuster, Pilar; Fisiología; Facultades de la UMU
    Objective: To detect plasminogen and plasminogen activators (PA) in oviduct and oocytes and to clarify the role of the plasminogen/plasmin system on mammalian fertilization. Design: Experimental prospective study. Setting: Mammalian reproduction research laboratory. Animal(s): Oviducts and ovaries from porcine and bovine females were collected at slaughterhouse. A total of 52 oviducts and 2,292 oocytes were used. Boar and bull ejaculated spermatozoa were also used. Intervention(s): Plasminogen concentration in oviductal fluid (OF) through the cycle was measured. Immunolocalization of plasminogen and PAs in oocytes was carried out before and after fertilization. Porcine and bovine oocytes were in vitro fertilized, with plasminogen and plasmin added to the culture medium at different concentrations. Main Outcome Measure(s): Plasminogen concentration in OF. Plasminogen and PAs immunolocalization in oocytes. Penetration and monospermy rates, number of spermatozoa in the ooplasma and on the zona pellucida (ZP) after IVF. Result(s): Oviductal fluid contains about 92 mg/mL of plasminogen. The mature oocyte shows immunoreactivity toward plasminogen and toward PAs on its oolemma and ZP. After fertilization, plasminogen and PAs immunolabeling decreases in the oocyte, suggesting its conversion into plasmin. When exogenous plasminogen is added to the IVF medium, sperm entry into the oocyte is hampered, suggesting that the role of plasminogen activation during fertilization is to reduce the number of (or to select) penetrating spermatozoa. Conclusion(s): The plasminogen/plasmin system is activated during gamete interaction and regulates the sperm entry into the oocyte
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    Mimicking the temperature gradient between the sow’s oviduct and uterus improves in vitro embryo culture output
    (Oxford University Press, 2020-07-09) García Martínez, Soledad; Latorre Reviriego, Rafael Manuel; Sánchez-Hurtado, M. A.; Sánchez-Margallo, F. M.; Bernabò, N.; Romar Andrés, Raquel; López Albors, Octavio Miguel; Coy Fuster, Pilar; Anatomía y Anatomía Patológica Comparada
    This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21–168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.
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    How is plasminogen/plasmin system contributing to regulate sperm entry into the oocyte?
    (2013-12-30) Grullón LA; Gadea Mateos, Joaquín; Mondéjar Corbalán, Irene; Romar Andrés, Raquel; Matas Parra, Carmen; Coy Fuster, Pilar; Coy Fuster, Pilar; Fisiología; Facultades de la UMU
    Plasminogen is present in the oviduct, on the zona pellucida (ZP) and on oolemma, and reduces the number of sperm penetrating the oocyte during in vitro fertilization in pig and cow. It is unknown how this reduction occurs. We tested whether plasminogen (1) changed the ZP resistance to enzymatic digestion thus making the passage of the spermatozoa across it difficult; (2) reduced the sperm functionality, assessed by sperm viability, motility, spontaneous acrosome reaction and membrane lipid disorder; or (3) affected the sperm-ZP binding before or after sperm-ZP interaction. The mechanism by which plasminogen/plasmin system contributes to regulate sperm entry into the oocyte is not inducing a ZP hardening or a decrease in sperm functionality but detaching more than 50% of sperm bound to the ZP. It is suggested that the fertilizing spermatozoon activates plasminogen into plasmin at the oocyte surface and that plasmin removes additional spermatozoa attached to the ZP.
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