A single dose of allopregnanolone affects rat ovarian morphology and steroidogenesis

Allopregnanolone, a progesterone metabolite, is one of the best characterized neurosteroids. In a dose that mimics serum levels during stress, allopregnanolone inhibits sexual receptivity and ovulation and induces a decrease in luteinizing hormone levels. The aim of this work was to examine the effect of an intracerebroventricular administration of allopregnanolone on ovarian morphophysiology; serum and tissue levels of progesterone and estrogen; and enzymatic activity of 3β-hydroxysteroid dehydrogenase, 20α-hydroxysteroid dehydrogenase and 3α-hydroxysteroid oxido-reductase in the ovary and in the medial basal hypothalamus on the morning of estrus. Ovarian morphology was analyzed under light microscopy. The hormone assays were performed by radioimmunoassay. The enzymatic activities were measured by spectrophotometric analysis. The morphometric analysis revealed that, in allopregnanolone-treated animals, the number of secondary and Graafian follicles was decreased, whereas that of atretic follicles and cysts was significantly increased. Some cysts showed luteinized unruptured follicles. There were no differences in the number of tertiary follicles or corpora lutea in comparison with the corresponding control groups. In allopregnanolone-treated animals, progesterone serum levels were increased, whereas ovarian progesterone levels were decreased. Moreover, 3β-HSD and 3α-HSOR enzymatic activities were increased in the medial basal hypothalamus, whereas ovarian levels were decreased. The enzyme 20α-hydroxysteroid dehydrogenase showed the opposite profile. The results of this study showed that allopregnanolone interferes on ovarian steroidogenesis and ovarian morphophysiology in rats, providing a clear evidence for the role of this neurosteroid in the control of reproductive function under stress situations.Fil: Pelegrina, Laura Tatiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Cáceres Gimenez, Antonella Rosario Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Giuliani, Fernando Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Asensio, Joana Antonela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Parborell, Maria Fernanda Agustina. 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: Laconi, Myriam Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentin

Efecto de los 4-pregnenos (derivados de la progesterona) sobre el comportamiento biológico de líneas celulares derivadas tumores de ovario

Comunicación científica en formato ORAL, realizada en las III Jornadas Internacionales de Investigación, Ciencia y Universidad y las XII Jornadas de Investigación UMaza, en el Bloque de comunicaciones científicas: "BIOLOGÍA MOLECULAR, BIOQUÍMICA E INMUNOLOGÍA", el mismo fue moderado por el Dr. CRISTIAN QUINTERO. Las jornadas se llevaron adelante desde 19 al 23 de octubre del 2020 en formato totalmente virtual bajo plataforma Zoom y fueron transmitidas por el canal YouTube de la UMaza y el Facebook del Área de Ciencia y Técnica UMaza (Somos Ciencia y Técnica UMaza)

Efecto de los 4-pregnenos (derivados de la progesterona) sobre el comportamiento biológico de líneas celulares derivadas tumores de ovario

El cáncer de ovario es la forma más letal de neoplasia ginecológica y la quinta causa de muerte por esta enfermedad en mujeres. Alrededor de un 90% de los cánceres de ovario son de origen epitelial. En la actualidad existen evidencias controversiales acerca del rol de progesterona en la carcinogénesis ovárica. Esta hormona puede ser metabolizada en los derivados 4-preg nenos, 3α-di-hidroprogesterona (3HP) y 20α-dihidroprogesterona (20HP), o en los derivados 5α pregnanos (allopregnanolona). En cáncer de mama se ha demostrado que los derivados de la progesterona podían tener efectos pro-tumorales (5-pregnanos) o anti-tumorales (4-pregnenos). A su vez, existe una mayor producción de los derivados 5-pregnanos que de los 4-pregnenos en células y tejidos tumorigénicos de mama. Nosotros demostramos que allopregnanolona tiene un efecto pro-tumoral en dos líneas celulares derivadas de pa cientes con cáncer de ovario, IGROV-1 y SKOV-3

Caracterización histológica e histoquímica de branquia, hígado y riñón de perca criolla ( Percichthys trucha, Valenciennes, 1833 ) para su uso en biomonitoreo ambiental

ARTÍCULO PUBLICADO EN REVISTA EXTERNA. El objetivo de esta investigación fue realizar la descripción histológica de los principales órganos blanco de contaminantes ambientales en Percichthys trucha, pez nativo y ampliamente distribuido en ríos y lagos del sur de Argentina. Se capturaron sesenta especímenes adultos sanos de tamaño y peso similar, en Embalse El Nihuil, Mendoza, Argentina. Se describieron macroscópicamente las branquias, el hígado y el riñón y se fijaron en formalina neutra bufferada al 10% para su inclusión y tinción con hematoxilina eosina, ácido peryódico de Schiff (PAS), tricrómico de Masson y azul de Alcian. Se utilizó un microscopio trinocular con cámara y software de imagen para la evaluación microscópica. Cada branquia se compone de cuatro arcos con dos filamentos. Cada filamento está constituido por un núcleo de cartílago y una mucosa con repliegues (lamelas) de epitelio plano simple con células caliciformes PAS y Azul Alcian positivo. La posición y la distancia entre las lamelas mantienen la simetría, al igual que el grosor del epitelio. El hígado está encapsulado con tejido pancreático interpuesto. Los lóbulos hepáticos no están bien delimitados, pero se identifican las áreas portales. Las trabéculas están formadas por dos placas de hepatocitos con una cara sinusoidal para absorción y otra biliar para excreción. Los hepatocitos contienen grandes cantidades de lípidos y reservas de glucógeno PAS positivas. El riñón tiene nefronas bien desarrolladas. Se describen los centros melano-macrófagos. Se tomaron medidas microscópicas. Estos resultados sirven de referencia para determinar biomarcadores histopatológicos de contaminación acuática, herramienta útil y económica para el monitoreo biológico. Sitio de la revista: https://acta-microscopica.org/acta/article/view/12

Erratum: A single dose of allopregnanolone affects rat ovarian morphology and steroidogenesis((2017) 153 (75–83) DOI: 10.1530/REP-16-0463)

Allopregnanolone, a progesterone metabolite, is one of the best characterized neurosteroids. In a dose that mimics serum levels during stress, allopregnanolone inhibits sexual receptivity and ovulation and induces a decrease in luteinizing hormone levels. The aim of this work was to examine the effect of an intracerebroventricular administration of allopregnanolone on ovarian morphophysiology, serum and tissue levels of progesterone and estrogen, and enzymatic activity of 3β-hydroxysteroid dehydrogenase, 20α-hydroxysteroid dehydrogenase and 3α?hydroxysteroid oxido-reductase in the ovary and in the medial basal hypothalamus on the morning of estrus. Ovarian morphology was analyzed under light microscopy. The hormone assays were performed by radioimmunoassay. The enzymatic activities were measured by spectrophotometric analysis. The morphometric analysis revealed that, in allopregnanolone-treated animals, the number of secondary and Graafian follicles was decreased while that of atretic follicles and cysts was significantly increased. Some cysts showed luteinized unruptured follicles. 18 There were no differences in the number of tertiary follicles or corpora lutea in comparison with the corresponding control groups. In allopregnanolone-treated animals, progesterone serum levels were increased, while ovarian progesterone levels were decreased. Moreover, 3β-HSD and 3α-HSOR enzymatic activities were increased in the medial basal hypothalamus while ovarian levels were decreased. The enzyme 20α-hydroxysteroid dehydrogenase showed the opposite profile. The results of this study showed that allopregnanolone interferes on ovarian steroidogenesis and ovarian morphophysiology inrats, providing a clear evidence for the role of this neurosteroid in the control of reproductive function under stress situations.Fil: Pelegrina, Laura Tatiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Cáceres, Antonella Rosario Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Asensio, Joana Antonela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Parborell, Maria Fernanda Agustina. 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: Laconi, Myriam Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentin

Effects of short-term in vitro heat stress on bovine preantral follicles

Heat stress occurs when heat dissipation mechanisms are overwhelmed by external and internal heat production. Ovarian reserve is an important contributor to reproductive success in cattle. The effects of heat stress on antral follicle development have been well studied; the consequences on preantral follicles are still largely unknown. This study aimed to investigate and characterize the effects of a short in vitro heat stress exposure on bovine preantral follicles, with a special focus on primordial follicles. Bovine ovarian cortex fragments (3 × 3 × 1 mm) were isolated and incubated at different temperatures for 2 h. The incubation methodology was validated using a non-incubated group. Incubation groups consisted in physiological temperature (38.5 °C) or heat stress (41 °C). Data on follicle count, diameter of the primordial follicles, number of granulosa cells, and markers for follicle proliferation and degeneration were recorded. Heat stress caused a significant decrease in the number of primordial follicles (p < 0.0001) compared to control and non-incubated groups. The diameter of these follicles also decreased under heat stress (p < 0.0001), as did the number of granulosa cells. In addition, an increase in the number of degenerated follicles (p < 0.0001) and a decrease in granulosa cell proliferation (p < 0.0001) were found after exposure to heat stress. The results of this in vitro study show that a brief exposure to heat stress can affect the follicular reserve in the ovarian cortex and therefore, provide relevant information of its effects on the reproductive performance of cattle.Fil: Cardone, Daniela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad de Mendoza; ArgentinaFil: Cáceres Gimenez, Antonella Rosario Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad de Mendoza; ArgentinaFil: Sanhueza, María de Los Ángeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Bruna, Flavia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Laconi, Myriam Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad de Mendoza; Argentin

Effect of progesterone and first evidence about allopregnanolone action on the progression of epithelial human ovarian cancer cell lines

ARTÍCULO PUBLICADO EN REVISTA EXTERNA. Ovarian carcinoma is one of the most common cause of death by gynecologic cancer. Several epidemiological and in vitro studies have shown controversial data about progesterone effects in ovarian cancer. Progesterone can be converted in its active metabolite, allopregnanolone, its effects in ovarian cancer are still unknown. Previously, we demonstrated that allopregnanolone modifies ovarian morphophysiology, being able to alter critical process of tumor development such as proliferation, apoptosis and angiogenesis. Taking into account these antecedents, we investigated the effect of progesterone and allopregnanolone on proliferation, apoptosis, clonogenic capacity and migration on two epithelial human ovarian cancer cell lines, IGROV-1 and SKOV-3. To this end, IGROV-1 and SKOV-3 cells were exposed to a range of progesterone and allopregnanolone concentrations (10-11 to 10-5 M) for 72 h. Proliferation was analyzed by MTT and Ki67 expression. Apoptosis was measured by immunocytochemistry of cleaved caspase 3. Clonogenic capacity was evaluated by counting colonies. Migration was analyzed by wound assay. We found that allopregnanolone increased proliferation and Ki67 expression respect to control on IGROV-1 cells, while expression of cleaved caspase 3 did not change in any cell line studied. IGROV-1 clonogenic capacity was also increased by allopregnanolone treatment. Both steroids, progesterone and allopregnanolone, increased IGROV-1 migration in a concentration dependent manner. None of the steroids tested modified SKOV-3 biological behavior analized. This is the first evidence that allopregnanolone, a progesterone metabolite, affects critical events in tumor development of human epithelial ovarian cancer. These results could have an impact in the future in clinic diagnosis, prognosis and treatment of ovarian cancer patients. The regulation of progesterone and allopregnanolone steroideogenesis and their molecular mechanisms might be considered as potential therapeutic tool in ovarian cancer. Sitio de la revista: https://www.sciencedirect.com/science/article/pii/S0960076019301013?via%3Dihu

Progesterone has a neuroprotective effect and prevents depression sings in a male rat depression like model

Depression is one of the psychiatric disorders with the highest incidence in recent decades. It has been recently related to different neurodegenerative diseases such as Parkinson. These pathologies often appear associated with a premotor sign such as depression and decreased cognitive performance. The objective was to study and evaluate if the treatment with the neuroactive steroid progesterone could prevent the development of premotor early signs of neurodegenerative diseases in a model of catecholaminergic depletion by reserpine. Sprague-Dawley male rats (250-350 g), between 60 and 90 days old were used. The experimental groups were: C (saline), R (reserpine 0.1 mg/kg/sc, 10 injection over the course of 20 days), P (progesterone 4 mg/kg/sc) alone, 5 days after de experiments andPP + R (reserpine 5 days after a previous dose of progesterone). During the course of reserpine treatment, the animals were evaluated in the catalepsy test. Forced swimming and novel object recognition were tested before the appearance of the motor signs. Data were analyzed by ANOVA-1 and Tukey`s post hoc. A significant decrease (p<0.05) was observed between the R vs C groups in all evaluated behavioral parameters. There were no significant differences in the catalepsy time in the PP+R vs C. Although PP+R vs R showed a significant increase in the time spent swimming in the forced swimming test (p <0.05) and in the discrimination index (p <0.05) in novel object recognition. We conclude that a previous progesterone treatment can avoid depression-like behavior and improves short time memory and the on/ off effect on locomotor activity. Progesterone exerts a neuroprotective effect against the reserpine treatment, preventing cognitive and depression premotor disorders induced by catecholamine depletion.Fil: Villegas, Vanina Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Mulle Bernedo, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: García, Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Cáceres Gimenez, Antonella Rosario Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Yunes, Roberto Miguel Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Cabrera Kreiker, Ricardo Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentin

Allopregnanolone alters follicular and luteal dynamics during the estrous cycle

Background: Allopregnanolone is a neurosteroid synthesized in the central nervous system independently of steroidogenic glands; it influences sexual behavior and anxiety. The aim of this work is to evaluate the indirect effect of a single pharmacological dose of allopregnanolone on important processes related to normal ovarian function, such as folliculogenesis, angiogenesis and luteolysis, and to study the corresponding changes in endocrine profile and enzymatic activity over 4 days of the rat estrous cycle. We test the hypothesis that allopregnanolone may trigger hypothalamus - hypophysis - ovarian axis dysregulation and cause ovarian failure which affects the next estrous cycle stages. Methods: Allopregnanolone was injected during the proestrous morning and then, the animals were sacrificed at each stage of the estrous cycle. Ovarian sections were processed to determine the number and diameter of different ovarian structures. Cleaved caspase 3, proliferating cell nuclear antigen, α-actin and Von Willebrand factor expressions were evaluated by immunohistochemistry. Luteinizing hormone, prolactin, estrogen and progesterone serum levels were measured by radioimmunoassay. The enzymatic activities of 3β-hydroxysteroid dehydrogenase, 3α-hydroxysteroid oxidoreductase and 20α-hydroxysteroid dehydrogenase were determined by spectrophotometric assays. Two-way ANOVA followed by Bonferroni was performed to determine statistical differences between control and treated groups along the four stages of the cycle. Results: The results indicate that allopregnanolone allopregnanolone decreased the number of developing follicles, while atretic follicles and cysts increased with no effects on normal cyclicity. Some cysts in treated ovaries showed morphological characteristics similar to luteinized unruptured follicles. The apoptosis/proliferation balance increased in follicles from treated rats. The endocrine profile was altered at different stages of the estrous cycle of treated rats. The angiogenic markers expression increased in treated ovaries. As regards corpora lutea, the apoptosis/proliferation balance and 20α-hydroxysteroid dehydrogenase enzymatic activity decreased significantly. Progesterone levels and 3β-hydroxysteroid dehydrogenase enzymatic activity increased in treated rats. These data suggest that allopregnanolone interferes with steroidogenesis and folliculogenesis at different stages of the cycle. Conclusion: Allopregnanolone interferes with corpora lutea regression, which might indicate that this neurosteroid exerts a protective role over the luteal cells and prevents them from luteolysis. Allopregnanolone plays an important role in the ovarian pathophysiology.Fil: Asensio, Joana Antonela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Cáceres Gimenez, Antonella Rosario Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad "Juan Agustín Maza". Facultad de Ciencias Veterinarias y Ambientales; ArgentinaFil: Pelegrina, Laura Tatiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina. Universidad "Juan Agustín Maza". Facultad de Ciencias Veterinarias y Ambientales; ArgentinaFil: Sanhueza, María de Los Ángeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Scotti, Leopoldina. 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: Parborell, Maria Fernanda Agustina. 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: Laconi, Myriam Raquel. Universidad "Juan Agustín Maza". Facultad de Ciencias Veterinarias y Ambientales; Argentina. Universidad de Mendoza; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentin