Editorial: 3D Modelling of Mammalian Embryos and Organs

The main scope of this Special issue was to gain understanding on how tissues and organs are arranged into integrative hierarchical levels of complexity, from the molecular to the morphological organization. To understand the underlying complexity of the relationships among these levels during morphogenesis or in the adult we must learn how to resolve single-cell spatial relationships in the three-dimensional (3D) organisation of tissues, organs and even of the whole organisms.Fil: Garagna, Silvia. Universita Degli Studi Di Pavia; ItaliaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Aréchaga, Juan. Universidad del País Vasco; EspañaFil: Zuccotti, Maurizio. Universita Degli Studi Di Pavia; Itali

Early Abnormal Placentation and Evidence of Vascular Endothelial Growth Factor System Dysregulation at the Feto-Maternal Interface After Periconceptional Alcohol Consumption

Adequate placentation, placental tissue remodeling and vascularization is essential for the success of gestation and optimal fetal growth. Recently, it was suggested that abnormal placenta induced by maternal alcohol consumption may participate in fetal growth restriction and relevant clinical manifestations of the Fetal Alcohol Spectrum Disorders (FASD). Particularly, periconceptional alcohol consumption up to early gestation can alter placentation and angiogenesis that persists in pregnancy beyond the exposure period. Experimental evidence suggests that abnormal placenta following maternal alcohol intake is associated with insufficient vascularization and defective trophoblast development, growth and function in early gestation. Accumulated data indicate that impaired vascular endothelial growth factor (VEGF) system, including their downstream effectors, the nitric oxide (NO) and metalloproteinases (MMPs), is a pivotal spatio-temporal altered mechanism underlying the early placental vascular alterations induced by maternal alcohol consumption. In this review we propose that the periconceptional alcohol intake up to early organogenesis (first trimester) alters the VEGF-NO-MMPs system in trophoblastic-decidual tissues, generating imbalances in the trophoblastic proliferation/apoptosis, insufficient trophoblastic development, differentiation and migration, deficient labyrinthine vascularization, and uncompleted remodelation and transformation of decidual spiral arterioles. Consequently, abnormal placenta with insufficiency blood perfusion, vasoconstriction and reduced labyrinthine blood exchange can be generated. Herein, we review emerging knowledge of abnormal placenta linked to pregnancy complications and FASD produced by gestational alcohol ingestion and provide evidence of the early abnormal placental angiogenesis-vascularization and growth associated to decidual-trophoblastic dysregulation of VEGF system after periconceptional alcohol consumption up to mid-gestation, in a mouse model.Fil: Gualdoni, Gisela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Jacobo, Patricia Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Barril, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Ventureira, Martín Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Abnormal growth and morphogenesis of placenta at term is linked to adverse fetal development after perigestational alcohol consumption up to early gestation in mouse

Background: Gestation alcohol consumption produces fetal growth restriction and malformations by affecting the embryo–fetal development. Recently a relationship between abnormal placentation and fetal malformation and intrauterine growth retardation has been suggested. However, the effects of perigestational alcohol ingestion up to early pregnancy on the placenta at term and its association with fetal abnormalities are little known. Methods: In female mice, ethanol 10% in water was administered for 15 days previous and up to days 4 (D4), 8 (D8), or 10 (D10) of gestation (TF), and gestation continues without ethanol exposure. Control females (CF) received ethanol-free water. At day 18, feto-placental units and implantation sites were studied. Results: TF had increased resorptions and only fetuses from D8-TF and D10-TF had significantly increased weights versus CF. D4 and D10-TF-placentas had significantly reduced weights. All TF had increased junctional zone (JZ) and reduced labyrinth (Lab) areas (PAS-histology and morphometry) compared with CF. Fetuses with mainly with craniofacial abnormalities and skeletal defects (Alizarin red staining), significantly increase; while the fetal bone density (alizarin color intensity, ImageJ) was reduced in D4, D8 and D10-TF versus CF. Although all TF-placentas were histo-structural affected, TF-abnormal fetuses had the most severe placental anomalies, with junctional abundant glycogenic cells into the labyrinth, disorganized labyrinthine vascularization with signs of leukocyte infiltrates and feto-maternal blood mix. Conclusions: Perigestational alcohol consumption up to early gestation induces at term fetal growth alterations, dysmorphology and defective skeleton, linked to deficient growth and abnormal morphogenesis of placenta, highlighting insight into the prenatal etiology of FASD.Fil: Gualdoni, Gisela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Pérez Tito, Leticia Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Barril, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Sobarzo, Cristian Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentin

Differential expression and activity of matrix metalloproteinases 2 and 9 in canine early placenta

The zonary and endotheliochorial dog placenta is the most invasive placenta of carnivores. The importance of matrix metalloproteinases (MMP) in placenta invasiveness has been determined in several mammals including species with haemochorial, epitheliochorial and endotheliochorial placentation. Regarding the latter, the expression of MMP enzymes has been studied in the cat and the mature canine placenta. The aim of this study was to analyse the expression and activity of MMP-2 and MMP-9 in the early dog placenta. Placentae from 18 to 30 days of pregnancy were collected from four bitches. Two placentae from each bitch were analysed. Placental tissue from one uterine horn was fixed in formaldehyde for immunohistochemistry, while marginal haematoma, labyrinth, non-implantative and implantative endometrium from the contralateral horn were immediately frozen in dry ice for the analysis of MMP expression (Western blot [WB]) and activity (zymography). MMP-2 and MMP-9 were evidenced in the labyrinth, maternal glands and marginal haematoma; this finding was directly correlated with levels of MMP expression by WB, and with the activity of MMP-2, mainly in the haematoma (the area of major remodelling of tissues). Thus, although MMP-9 is well expressed in the early canine placenta, it is not active. Given the important role of MMPs for invasiveness, maternal–foetal angiogenesis and the establishment of a correct foetal nutrition, the results are consistent with the findings in other species in which the MMP-2 activation precedes the MMP-9 one in early placentation.Facultad de Ciencias Veterinaria

El consumo semicrónico moderado de alcohol altera la foliculogénesis y la calidad núcleo-citoplasmática oocitaria, en el ratón

El consumo crónico de alcohol es un problema de salud mundial que afecta particularmente a la población femenina. Sin embargo, los efectos de la ingesta semicrónica en cantidades moderadas a bajas en el ovario y el oocito son poco conocidos. En un modelo murino, se administró etanol al 10% en agua de bebida (hembras tratadas) o agua (hembras control) por 15 días, y luego de la superovulación o no (ovulación espontánea), se analizó el ciclo estral y la calidad ovárico-gamética. En las hembras tratadas, la frecuencia y duración del diestro aumentó, y las frecuencias de folículos y cuerpos lúteos disminuyeron vs hembras controles, valores que se restauraron luego de la superovulación. Sin embargo, en las hembras tratadas, la tasa de proliferación celular folicular y el desbalance de la expresión ovárica de VEGF (factor de crecimiento endotelial) persistieron luego de la superovulación. El número de ovocitos ovulados con metafase II anormal, fragmentados y activados partenogenéticamente fue mayor en las hembras tratadas respecto las controles. En conclusión, el consumo semicrónico moderado de alcohol produce anestro, ciclo estral irregular, foliculogénesis deficiente y anomalías núcleo-citoplasmáticas en los oocitos ovulados. Estas alteraciones podrían constituirse en un factor etiológico de pérdida gestacional temprana y desarrollo embrionario anormal luego del consumo de alcohol.Chronic alcohol consumption is a global health problem that particularly affects the female population. However, the effects of semi-chronic ethanol intake in low-moderate amounts on the ovary and oocyte are poorly understood. In a mouse model, 10% ethanol was administered in drinking water (treated females) or water (control females) for 15 days, and after superovulation or not (spontaneous ovulation), the estrous cycle and ovarian-gametic quality were analyzed. In treated females, the frequency and duration of the diestrus increased, and the frequencies of follicles and corpus luteum decreased vs control females, values that restored after superovulation. However, in treated females, the follicular cell proliferation rate and the imbalance in ovarian expression of VEGF (endothelial growth factor) persisted after superovulation. The number of ovulated oocytes with abnormal metaphase II, fragmented and parthenogenetically activated was higher in treated females than in control ones. In conclusion, moderate semi-chronic alcohol consumption produces anestrum, irregular estrous cycle, poor folliculogenesis, and nuclearcytoplasmic abnormalities in ovulated oocytes. These alterations could constitute an etiological factor of early gestational loss and abnormal embryonic development after alcohol consumption.Fil: Gualdoni, Gisela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Barril, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Jacobo, Patricia Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentin

Perigestational alcohol consumption induces altered early placentation and organogenic embryo growth restriction by disruption of trophoblast angiogenic factors

Research question: Maternal alcohol consumption produces fetal retardation and malformations, probably associated with placental defects. Does perigestational alcohol consumption up to organogenesis lead to abnormal placentation and embryo growth restriction by disrupting the vascular endothelial growth factor (VEGF) system in embryo–placental development? Design: Female mice were treated with 10% ethanol in drinking water before and up to day 10 of gestation. Control mice received ethanol-free water. After treatment, the trophoblastic tissue, embryo growth and the angiogenic VEGF pathway were analysed. Results: Female mice who had received treatment had resorbed and delayed implantation sites with poor ectoplacental cone development. Reduced trophoblastic area tissue from female mice who had received treatment had abnormal junctional zone and diminished labyrinthine vascularization. After treatment, the labyrinth had increased chorionic trophoblast proliferation, hypoxia inducible factor-1α immunoexpression but reduced apoptosis. The embryo growth was reduced concomitantly with low VEGF immunostaining but high endothelial nitric oxide synthase (eNOS) expression. In junctional and labyrinth of treated female mice, gene and protein immunoexpression of VEGF was reduced and the protein expression of FLT-1 increased compared with controls. Increased activation of kinase insert domain receptor receptor (phosphorylated KDR) and expression of eNOS were observed in placenta of treated female mice. Immunoexpression of metalloproteinase-9, however, was reduced in junctional zone but increased in labyrinth, compared with controls. Conclusions: These data reveal inadequate expression of VEGF/receptors and angiogenic eNOS and metalloproteinase factors related to abnormal early placentation after perigestational alcohol ingestion, providing insight into aetiological factors underlying early placentopathy associated with intrauterine growth restriction caused by maternal alcohol consumption.Fil: Gualdoni, Gisela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Ventureira, Martín Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Coll, Tamara Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad Nacional de San Martín. Instituto de Investigación en Ingeniería Ambiental; ArgentinaFil: Palomino, Wilder Alberto. Universidad de Santiago de Chile. Hospital Clinico San Borja Arriaran. Instituto de Investigaciones Materno Infantil; ChileFil: Barbeito, Claudio Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Involvement of metalloproteinase and nitric oxide synthase/nitric oxide mechanisms in early decidual angiogenesis–vascularization of normal and experimental pathological mouse placenta related to maternal alcohol exposure

Successful pregnancy for optimal fetal growth requires adequate early angiogenesis and remodeling of decidual spiral arterioles during placentation. Prior to the initiation of invasion and endothelial replacement by trophoblasts, interactions between decidual stromal cells and maternal leukocytes, such as uterine natural killer cells and macrophages, play crucial roles in the processes of early maternal vascularization, such as proliferation, apoptosis, migration, differentiation, and matrix and vessel remodeling. These placental angiogenic events are highly dependent on the coordination of several mechanisms at the early maternal–fetal interface, and one of them is the expression and activity of matrix metalloproteinases (MMPs) and endothelial nitric oxide synthases (NOSs). Inadequate balances of MMPs and nitric oxide (NO) are involved in several placentopathies and pregnancy complications. Since alcohol consumption during gestation can affect fetal growth associated with abnormal placental development, recently, we showed, in a mouse model, that perigestational alcohol consumption up to organogenesis induces fetal malformations related to deficient growth and vascular morphogenesis of the placenta at term. In this review, we summarize the current knowledge of the early processes of maternal vascularization that lead to the formation of the definitive placenta and the roles of angiogenic MMP and NOS/NO mechanisms during normal and altered early gestation in mice. Then, we propose hypothetical defective decidual cellular and MMP and NOS/NO mechanisms involved in abnormal decidual vascularization induced by perigestational alcohol consumption in an experimental mouse model. This review highlights the important roles of decidual cells and their MMP and NOS balances in the physiological and pathophysiological early maternal angiogenesis–vascularization during placentation in mice

Citotoxicity by chronic alcohol consumption on mouse gametes, fertilization and preimplantation embryogenesis

Se ha descripto que la ingesta crónica y/o aguda de altas concentraciones de etanol durante la gestación produce el síndrome de alcoholismo fetal (FAS). El objetivo principal de este estudio fue profundizar los conocimientos sobre los efectos deletéreos del consumo preconcepcional de etanol y sus consecuencias sobre el desarrollo preimplantativo del embrión. Machos adultos y hembras prepúberes de ratón Fl de (C57 x CBA), ingirieron etanol al 10, 5 y 2.5% en el agua de bebida por 30 días. A los 27 días del tratamiento se indujo la ovulación. Se estudio la calidad de las gametas, la tasa de fecundación y el grado de desarrollo preimplantativo alcanzado por embriones in vitro e in vivo (hasta dia 4), y adicionalmente la incidencia del consumo de etanol en la maduración sexual de las hembras. La administración crónica de etanol al 10 y 5% produjo retraso en la maduración folicular. Como consecuencia, los ovocitos ovulados presentaron altos índices de activación espontánea, partenogenética, y la capacidad de sintesis de PGE por el complejo cúmulo-ovocitario fue reducida con el consumo del 10 y aumentada con el consumo del 5% de etanol. La tasa de fecundación in vitro e in vivo fue reducida cuando las hembras fueron tratadas con ambas dosis de etanol. Los ovocitos intactos de las hembras tratadas no fueron penetrados por los espermatozoides, y los ovocitos haploides oviductales permanecieron sin ser fecundados. El etanol afectó tanto la maduración del ovocito, la regulación y control de su calidad por las células del cúmulo, y también posiblemente la membrana plasmática ovocitaria y otros componentes celulares, lo que resultó en la pérdida de fecundabílidad. El consumo crónico de etanol al lO y 5%, en la fase preconcepcional de las hembras, produjo, tanto en el desarrollo in vitro como in vivo, tasa embrionaria diaria reducida, debido al crecimiento retrasado, al arresto embrionario en estadios de mórulas y en blastocistos tempranos, en los días 2, 3 y 4 (in vivo) y 5 de desarrollo in vitro, con ambas dosis de etanol. Asimismo, fue evidente la alta tasa de fragmentación embrionaria temprana, desde día 2 con etanol al 10%, y menor y más tardía en las hembras tratadas con etanol al 5%. El consumo de etanol al 10% produjo aumento de embriones anormales. El impacto del alcohol en el desarrollo embrionario fue menor con la dosis del 5%, debido a retraso y pérdida embrionaria de reducida intensidad. Se puede concluir que la dosis mínima perjudicial para la gameta femenina y el desarrollo ulterior del embrión preimplantativo, administrada cronicamente a hembras prepúberes, fue el etanol al 5%. Los efectos del consumo crónico de etanol al 5% podrian deberse en parte a las acciones "subcrónicas" del etanol en los días periovulatorios.It was described that the chronic or acute consumption of high doses of ethanol during gestation produces the fetal alcohol syndrome (FAS). The main objetive of this study was to progress in the aknowledgement about the deleterious effects of preconceptional consumption of ethanol and the consequences on the preimplantation embryo development. Adult male and prepubertal female mice (C57 x CBA) F1), were treated with 10,5 and 2.5 % w/v of ethanol in drinking water for 30 days. On day 27 of the treatment, females were induce to superovulate. The quality of the gametes, the fertilization rates, the stages of preimplantation development reached by the embryos in vitro and in vivo (up to day 4) were studied, and the incidence of the ethanol ingestion on the sexual maturation in the female was determined. The chronic administration of 10 and 5% of ethanol produced delayed follicular maturation. In consequence, the ovulated oocytes showed increased levels of spontaneous parthenogenetic activation, and the capacity of PGE synthesis by cumulus-oocyte complex (COC) was reduced by the 10% ethanol intake and increased by the 5% ethanol. The in vitro and in vivo fertilization rate was only reduced when the females were treated with both doses of ethanol. The intact oocytes from the treated females were not penetrated by spermatozoa, and the increased oviductal haploid oocytes stayed unfertilized. The ethanol affected the maturation of the oocyte, the regulation and the control of the oocyte quality by the cumulus cells, also probably the oolema and others celular components, which resulted in fertility loss. The chronic consumption of 10 and 5% of ethanol by females in the preconceptional phase produced, in both in vitro and in vivo development, reduced daily rates of embryos, due to delayed growth, to embryonary arrest in morulae and early blastocyst stages, in days 2, 3 and 4 (in vivo) and 5 (in vitro) of the development, with both doses of ethanol. Also, it was evident high early embryo fragmentation rates, from day 2 with ethanol 10%, and lower and later in females treated with 5% of ethanol. The ingestion of 10% of ethanol produced increase of abnormal embryos. The ethanol impact in the embryo development was lower with the 5%, due to the reduced intensity of delayed growth and embryo loss. It was concluded that the lower deletereous dose of ethanol, administered chronically to prepubertal females, that affects the gamete an the ulterior development of preimplantation embryos, was the 5% of ethanol. The effects of the chronic alcohol consumption could due to, in part, the "subchronic" actions of the ethanol on the periovulatory days.Fil:Cebral, Elisa. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

The role of oocyte morphology in the early embryo competence determination

El complejo proceso del desarrollo, tanto del embrión, del feto como de la placenta, comienza en las primeras fases de la gestación, durante la preimplantación. Sin embargo, el destino del embrión de mamíferos está marcado celular y molecularmente, por una red de señales génicas claves durante el desarrollo del oocito, cuando éste adquiere la competencia para el desarrollo embrionario. Esta nueva progenie se relacionará directamente con la morfogénesis normal del oocito, proceso íntimamente ligado al mapa molecular específico de expresión de proteínas y genes, que dirigen y regulan el desarrollo morfológico normal del oocito. Nuevos marcadores génicos de competencia oocito-embrionaria aportarán la posibilidad de monitoreo de las causas de bajas tasas de embarazo, de la disminución de la fertilidad, de aborto o pérdida temprana de la gestación o de anomalías embrionarias, ocasionadas por múltiples factores, uno de ellos la ingesta materna de alcohol. Esta revisión pretende brindar conocimiento básico acerca del desarrollo morfológico clásico del oocito y del embrión preimplantativo, y acercar un nuevo concepto, amplio y complejo, sobre la relación entre el aspecto morfológico clásico del oocito y del embrión con la “morfología molecular” de la gameta femenina, que determinan la competencia y el destino del embrión en desarrollo. Así, la vinculación entre la morfología oocitaria, la competencia nuclear y marcadores moleculares específicos podrían constituirse en útiles y esenciales herramientas para la determinación del destino embrionario, y con ello, potenciales elementos para el tratamiento de la pérdida temprana de la gestación.The complex process of development, the embryo, the fetus and placenta, begins in the early stages of pregnancy, during preimplantation. However, the fate of the mammalian embryo is marked at cellular and molecular level, by a network of key gene signals during oocyte development, when acquiring competition for embryonic development. This new progeny will be directly related to normal morphogenesis of the oocyte, a process intimately related to specific molecular map of proteins and genes expression, which directs and regulates normal morphological development of the oocyte. New gene markers of oocyte-embryonic competition provide the possibility of monitoring the causes of low pregnancy rates, of decreased fertility, abortion or early loss of gestation or embryonic anomalies, caused by multiple factors, one of them the maternal alcohol intake. This review aims to provide basic knowledge of classical morphological development of the oocyte and preimplantational embryo, and bring new concept, large and complex, on the relationship between the morphologic classic appearance of the oocyte and the embryo with the "molecular morphology" of the female gamete, determining the jurisdiction and the fate of the developing embryo. Thus, the relationship between oocyte morphology, nuclear competition and specific molecular markers could constitute useful and essential tool for determining the embryonic target, and thus potential components for the treatment of early pregnancy loss.Sociedad de Ciencias Morfológicas de La Plat

Successful capacitation and homologous fertilization in vitro in Calomys musculinus and Calomys laucha (Rodentia, Sigmodontinae)

Small South American rodents of the genus Calomys have been used extensively for virology and ecological research. Previous studies have demonstrated that Calomys musculinus and Calomys laucha have a relatively short oestrous cycle and that superovulation and parthenogenetic activation can be induced. The purpose of this study was to determine the requirements for in vitro manipulation of the male gamete and in vitro fertilization. Two culture media and different concentrations of spermatozoa were tested for their ability to support sperm motility, hyperactivation and the acrosome reaction. The ability of capacitated Calomys spermatozoa to penetrate zona-free hamster eggs was also evaluated. In vitro fertilization was assessed by examining attachment and binding to the zona pellucida, second polar body extrusion, pronucleus formation and the fertilizing sperm tail. The results of the study showed that: (i) Tyrode’s albumin lactate pyruvate (TALP) medium was more effective than T6 medium for maintaining sperm motility in vitro; (ii) hyperactivation was achieved with TALP but not with T6; (iii) the acrosome reaction was easily distinguished by light microscopy and depends on time and sperm concentration; (iv) capacitated spermatozoa are able to penetrate zona-free hamster eggs; and (v) superovulated oocytes can be fertilized in vitro. This is the first report of capacitation and in vitro fertilization for Calomys sp. These results provide opportunities to use C. musculinus and C. laucha as new laboratory animals for research into reproductive biology.Fil: Lasserre, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Cebral, Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Vitullo, Alfredo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentin