Romar Andrés, Raquel

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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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Open Access
Metabolic profile and glycemic response in fully-grown sows born using assisted reproductive technologies
(Elsevier, 2024-10-02) Cánovas Bernabé, Sebastián; Heras García, Sonia; Romero Aguirregomezcorta, Jon; Quintero Moreno, Armando Arturo; Gadea Mateos, Joaquín; Coy Fuster, Pilar; Romar Andrés, Raquel; Anatomía y Anatomía Patológica Comparada; Facultad de Veterinaria
The aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (P < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.
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Oviduct-specific glycoprotein and heparin modulate sperm-zona pellucida interaction during fertilization and contribute to the control of polyspermy
(2008-10-14) Saavedra, MD; Grullón, LA; Coy Fuster, Pilar; Cánovas Bernabé, Sebastián; Mondéjar Corbalán, Irene; Romar Andrés, Raquel; Matas Parra, Carmen; Avilés Sánchez, Manuel; Fisiología
Polyspermy is an important anomaly of fertilization in placental mammals, causing premature death of the embryo. It is especially frequent under in vitro conditions, complicating the successful generation of viable embryos. A block to polyspermy develops as a result of changes after sperm entry (i.e., cortical granule exocytosis). However, additional factors may play an important role in regulating polyspermy by acting on gametes before sperm-oocyte interaction. Most studies have used rodents as models, but ungulates may differ in mechanisms preventing polyspermy. We hypothesize that zona pellucida (ZP) changes during transit of the oocyte along the oviductal ampulla modulate the interaction with spermatozoa, contributing to the regulation of polyspermy. We report here that periovulatory oviductal fluid (OF) from sows and heifers increases (both, con- and heterospecifically) ZP resistance to digestion with pronase (a parameter commonly used to measure the block to polyspermy), changing from digestion times of approximately 1 min (pig) or 2 min (cattle) to 45 min (pig) or several hours (cattle). Exposure of oocytes to OF increases monospermy after in vitro fertilization in both species, and in pigs, sperm-ZP binding decreases. The resistance of OF-exposed oocytes to pronase was abolished by exposure to heparin-depleted medium; in a medium with heparin it was not altered. Proteomic analysis of the content released in the heparin-depleted medium after removal of OF-exposed oocytes allowed the isolation and identification of oviduct-specific glycoprotein. Thus, an oviduct-specific glycoprotein-heparin protein complex seems to be responsible for ZP changes in the oviduct before fertilization, affecting sperm binding and contributing to the regulation of polyspermy.
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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
Open Access
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.
Open Access
Differences in weight, hierarchy, and incidence of lameness between two groups of adult pigs derived from assisted reproductive technologies
(MDPI, 2022-12-17) Ramírez, Lisette L.; Ortin Bustillo, Alba; Ramis, Guillermo; Romar, Raquel; Coy, Pilar; Romero Aguirregomezcorta, Jon; Ramírez, Lisette L.; Ortín Bustillo, Alba; Ramis Vidal, Manuel Guillermo; Romar Andrés, Raquel; Coy Fuster, Pilar; Anatomía y Anatomía Patológica Comparadas ; Facultad de Veterinaria
The in vitro production (IVP) and subsequent transfer of embryos (ET) to recipient mothers is not yet an established reproductive technology in the pig industry, as it is in cattle. However, that the trade of IVP-cryopreserved pig embryos is expected to start in the next decades. Society and governments are increasingly aware of the repercussions that IVP could have for animal health, welfare, behavior, or food safety, but proven scientific information for this type of animal does not exist, since no colonies of pigs have been created to this end. We created a small one and studied the differences between 16 IVP-derived pigs and 14 pigs derived from artificial insemination (AI), at 3.5 years of age, conceived from the same boar, and housed and fed under the same conditions since they were born. Incidence of lameness, position in the herd hierarchy, weight, adenosine deaminase activity, and hematological and biochemical analytes were compared between the two groups of animals. The results showed that the IVP animals weighed more, occupied higher positions in the herd hierarchy, and had a lower incidence of lameness. Although genetic differences from the maternal line could explain some of these results, it is also possible that the IVP animals developed better adaptative abilities, but more studies with a higher number of animals are necessary to reach consistent conclusions.
Open Access
Growth parameters and growth-related hormone profile in a herd of cattle up to 4 years of age derived from assisted reproductive technologies
(MDPI, 2025-02-21) Heras García, Sonia; Sena Lopes, Jordana; Quintero Moreno, Armando Arturo; Romero Aguirregomezcorta, Jon; Cánovas Bernabé, Sebastián; Romar Andrés, Raquel; Coy Fuster, Pilar; Fisiología; Veterinaria
Assisted reproductive technologies (ART) are routinely used in livestock to generate animals of high genetic value. Despite representing an outstanding accomplishment, recent studies suggest differences in health, fertility, and gestational length of in vitro-produced compared to in vivo-derived animals. Currently, there are no data available on the long-term effects of ART on growth and development. This observational study aimed to understand the relationship between growth and growth-influencing hormones in a herd of cattle derived from artificial insemination (AI) or from in vitro-produced embryos either with BSA (C-IVP) or with reproductive fluids (RF-IVP) as a protein source in culture. Cortisol was associated positively with weight in AI and negatively with body length in males. Thyroxine decreased with age, and it was positively associated with thoracic circumference in RF-IVP. Insulin-like growth factor-1 was greater in RF-IVP than in C-IVP, and it was positively associated with body length and withers height. Growth hormone was greater in females than in males and RF-IVP compared to AI and C-IVP. In conclusion, we present here the first datasets on growth parameters and growth-influencing hormones in cattle from birth to 4 years of age without observing major evidence of differences depending on the embryo origin.
Embargo
Growth performance in pigs derived from in vitro produced embryos is enhanced compared to their artificial insemination-derived counterparts from birth to adulthood
(Elsevier, 2025-03-04) Heras García, Sonia; Soriano-Ubeda, Cristina; Quintero Moreno, Armando Arturo; Romero Aguirregomezcorta, Jon; Paris-Oller, Evelyne; Gadea Mateos, Joaquín; Romar Andrés, Raquel; Cánovas Bernabé, Sebastián; Coy Fuster, Pilar; Fisiología
Research suggests that assisted reproductive technologies may lead to adverse effects on the offspring. To date, long-term effects from fetal development through adulthood have been scarcely researched. In the present observational study, we aimed to describe growth (body weight, BW; crown-rump length, CRL; and average daily gain, ADG), and growth-related hormone levels (growth hormone, GH; insulin-like growth factor 1, IGF-1;thyroxine, T4; cortisol) of pigs derived from artificial insemination (AI) and from embryos produced in vitro under two different conditions, C-IVP, with Bovine Serum Albumin (BSA), and RF-IVP, with reproductive fluids and BSA, as protein source in culture, from birth to 5 years of age. In vitro-produced animals showed greater BW, CRL and ADG than AI. Additionally, C-IVP animals had greater BW, CRL and ADG than RF-IVP up to 6 months of age. Regarding hormones, GH concentration was greater in RF-IVP compared to AI and C-IVP. In males, IGF-1 levels of RF-IVP were greater than those of AI. Instead, T4 levels of AI males were greater than C-IVP and RFIVP. Males showed greater IGF-1 and GH than females, the latter only from 1 year old. Instead, females had greater T4 concentration than males. No differences in cortisol were observed between groups or sexes. In conclusion, this study sets reference values of growth and hormone concentrations for adult pigs that are missing in literature. Although the differences observed between the experimental groups were within physiological ranges, there was a greater growth performance in the IVP groups.
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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
Long-term study of physical, haematological, and biochemical parameters in cattle with different embryo origins
(MDPI, 2025-06-14) Serrano-Albal, María; Romero Aguirregomezcorta, Jon; Cánovas Bernabé, Sebastián; Heras García, Sonia; Gadea Mateos, Joaquín; Coy Fuster, Pilar; Romar Andrés, Raquel; Anatomía y Anatomía Patológica Comparada; Facultad de Veterinaria
Assisted reproductive technologies are vital in cattle breeding to improve genetic selection and productivity. While early-life differences between artificially inseminated (AI) and in vitro-produced (IVP) cattle have been studied, long-term physiological, haematological, and biochemical effects remain unclear. This observational study assessed AI and IVP cattle from 1.5 to 5 years of age to determine if early differences persist. IVP cattle were produced after the transfer of the embryo produced by supplementing (RF-IVP group) or not supplementing (C-IVP) the embryo culture with oviductal and uterine fluids. Physical evaluations showed body mass index increased until 3.5 years, while temperature and respiratory rate declined with age, with no significant differences between AI and IVP groups. Haematological analysis revealed age-related changes, including decreased red and white blood cell counts and increased mean corpuscular volume and haemoglobin. AI cattle had higher white blood cell counts than IVP groups. Sex significantly influenced many haematological variables. Biochemical analysis showed age-related increases in total protein, creatinine, and urea, and decreases in glucose and alkaline phosphatase. AI cattle had lower cholesterol and creatinine than IVP groups. Despite group differences, all values remained within normal ranges. Sex affected albumin, cholesterol, triglycerides, and creatine kinase. This study provides the first long-term haematological and biochemical reference values for cattle from different reproductive methods, showing that age is the main influencing factor and supporting IVP cattle as a viable alternative to AI in breeding programs.
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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.