Luteinizing hormone regulates inhibin-alpha subunit expression through multiple signaling pathways involving steroidogenic factor-1 and beta-catenin in the macaque corpus luteum

We employed different experimental model systems to define the role of GATA4, beta-catenin, and steroidogenic factor (SF-1) transcriptional factors in the regulation of monkey luteal inhibin secretion. Reverse transcription polymerase chain reactions and western blotting analyses show high expression of inhibin-alpha, GATA4, and beta-catenin in corpus luteum (CL) of the mid-luteal phase. Gonadotropin-releasing hormone receptor antagonist-induced luteolysis model suggested the significance of luteinizing hormone (LH) in regulating these transcriptional factors. Inducible cyclic AMP early repressor mRNA expression was detected in the CL and no change was observed in different stages of CL. Following amino acid sequence analysis, interaction between SF-1 and beta-catenin in mid-stage CL was verified by reciprocal co-immunoprecipitation experiments coupled to immunoblot analysis. Electrophoretic mobility shift analysis support the role of SF-1 in regulating luteal inhibin-alpha expression. Our results suggest a possible multiple crosstalk of Wnt, cAMP, and SF-1 in the regulation of luteal inhibin secretion.

Involvement of Src family of kinases and cAMP phosphodiesterase in the luteinizing hormone/chorionic gonadotropin receptor-mediated signaling in the corpus luteum of monkey

Background: In higher primates, during non-pregnant cycles, it is indisputable that circulating LH is essential for maintenance of corpus luteum (CL) function. On the other hand, during pregnancy, CL function gets rescued by the LH analogue, chorionic gonadotropin (CG). The molecular mechanisms involved in the control of luteal function during spontaneous luteolysis and rescue processes are not completely understood. Emerging evidence suggests that LH/CGR activation triggers proliferation and transformation of target cells by various signaling molecules as evident from studies demonstrating participation of Src family of tyrosine kinases (SFKs) and MAP kinases in hCG-mediated actions in Leydig cells. Since circulating LH concentration does not vary during luteal regression, it was hypothesized that decreased responsiveness of luteal cells to LH might occur due to changes in LH/CGR expression dynamics, modulation of SFKs or interference with steroid biosynthesis. Methods: Since, maintenance of structure and function of CL is dependent on the presence of functional LH/CGR its expression dynamics as well as mRNA and protein expressions of SFKs were determined throughout the luteal phase. Employing well characterized luteolysis and CL rescue animal models, activities of SFKs, cAMP phosphodiesterase (cAMP-PDE) and expression of SR-B1 (a membrane receptor associated with trafficking of cholesterol ester) were examined. Also, studies were carried out to investigate the mechanisms responsible for decline in progesterone biosynthesis in CL during the latter part of the non-pregnant cycle. Results and discussion: The decreased responsiveness of CL to LH during late luteal phase could not be accounted for by changes in LH/CGR mRNA levels, its transcript variants or protein. Results obtained employing model systems depicting different functional states of CL revealed increased activity of SFKs pSrc (Y-416)] and PDE as well as decreased expression of SR-B1correlating with initiation of spontaneous luteolysis. However, CG, by virtue of its heroic efforts, perhaps by inhibition of SFKs and PDE activation, prevents CL from undergoing regression during pregnancy. Conclusions: The results indicated participation of activated Src and increased activity of cAMP-PDE in the control of luteal function in vivo. That the exogenous hCG treatment caused decreased activation of Src and cAMP-PDE activity with increased circulating progesterone might explain the transient CL rescue that occurs during early pregnancy.

Single nucleotide polymorphisms in genes that are common targets of luteotropin and luteolysin in primate corpus luteum: Computational exploration

Luteal insufficiency affects fertility and hence study of mechanisms that regulate corpus luteum (CL) function is of prime importance to overcome infertility problems. Exploration of human genome sequence has helped to study the frequency of single nucleotide polymorphisms (SNPs). Clinical benefits of screening SNPs in infertility are being recognized well in recent times. Examining SNPs in genes associated with maintenance and regression of CL may help to understand unexplained luteal insufficiency and related infertility. Publicly available microarray gene expression databases reveal the global gene expression patterns in primate CL during the different functional state. We intend to explore computationally the deleterious SNPs of human genes reported to be common targets of luteolysin and luteotropin in primate CL Different computational algorithms were used to dissect out the functional significance of SNPs in the luteinizing hormone sensitive genes. The results raise the possibility that screening for SNPs might be integrated to evaluate luteal insufficiency associated with human female infertility for future studies. (C) 2012 Elsevier B.V. All rights reserved,

Studies on reactivation of regressing bonnet monkey corpus luteum on day 1 of menses: A pilot study

Studies on functional characteristics of the regressing primate corpus luteum (CL) to luteotrophic stimulus on day 1 of the non-fertile menstrual cycle are scarce. Recombinant human luteinizing hormone (rhLH) (20 IU/Kg BW; n = 10) or human chorionic gonadotropin (hCG) (180 IU; n = 6) were administered intravenously to female bonnet monkeys on day 1 of menses. Exogenous treatment of rhLH or hCG caused a significant increase in circulating progesterone (P4) levels 2-4 hours post treatment (P < 0.05). Lutectomy prior to onset of menses confirmed that CL is the site of the increased P4 concentrations. Increased levels of phosphorylated P44/42 MAPK, MKK3/6 activation and concomitant histological changes were observed within 4 hours in CL of monkeys receiving hCG treatment. The results from this study demonstrate the acute progesterone synthesizing capacity of regressing monkey CL after LH or hCG challenge. This has potential implications for interpreting the steroidogenic response after gonadotropin stimulation tests in the early follicular phase of the normal ovulatory and anovulatory women undergoing controlled ovarian stimulation protocols as part of assisted reproductive technology (ART) and in women with polycystic ovarian syndrome.

Computational interrogation of cis-regulatory elements of genes that are common targets of luteotropin and luteolysin in the primate corpus luteum

The rapid recent increase in microarray-based gene expression studies in the corpus luteum (CL) utilizing macaque models gathered increasing volume of data in publically accessible microarray expression databases. Examining gene pathways in different functional states of CL may help to understand the factors that control luteal function and hence human fertility. Co-regulation of genes in microarray experiments may imply common transcriptional regulation by sequence-specific DNA-binding transcriptional factors. We have computationally analyzed the transcription factor binding sites (TFBS) in a previously reported macaque luteal microarray gene set (n = 15) that are common targets of luteotropin (luteinizing hormone (LH) and human chorionic gonadotropin (hCG)) and luteolysin (prostaglandin (PG) F-2 alpha). This in silico approach can reveal transcriptional networks that control these important genes which are representative of the interplay between luteotropic and luteolytic factors in the control of luteal function. Our computational analyses revealed 6 matrix families whose binding sites are significantly over-represented in promoters of these genes. The roles of these factors are discussed, which might help to understand the transcriptional regulatory network in the control of luteal function. These factors might be promising experimental targets for investigation of human luteal insufficiency. (C) 2012 Elsevier B.V. All rights reserved.

Analysis of 20alpha-hydroxysteroid dehydrogenase expression in the corpus luteum of the buffalo cow: effect of prostaglandin F2-alpha treatment on circulating 20alpha-hydroxyprogesterone levels

Background: During female reproductive cycles, a rapid fall in circulating progesterone (P4) levels is one of the earliest events that occur during induced luteolysis in mammals. In rodents, it is well recognized that during luteolysis, P4 is catabolized to its inactive metabolite, 20alpha-hydroxyprogesterone (20alpha-OHP) by the action of 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) enzyme and involves transcription factor, Nur77. Studies have been carried out to examine expression of 20alpha-HSD and its activity in the corpus luteum (CL) of buffalo cow. Methods: The expression of 20alpha-HSD across different bovine tissues along with CL was examined by qPCR analysis. Circulating P4 levels were monitored before and during PGF2alpha treatment. Expression of 20alpha-HSD and Nur77 mRNA was determined in CL at different time points post PGF2alpha treatment in buffalo cows. The chromatographic separation of P4 and its metabolite, 20alpha-OHP, in rat and buffalo cow serum samples were performed on reverse phase HPLC system. To further support the findings, 20alpha-HSD enzyme activity was quantitated in cytosolic fraction of CL of both rat and buffalo cow. Results: Circulating P4 concentration declined rapidly in response to PGF2alpha treatment. HPLC analysis of serum samples did not reveal changes in circulating 20alpha-OHP levels in buffalo cows but serum from pseudo pregnant rats receiving PGF2alpha treatment showed an increased 20alpha-OHP level at 24 h post treatment with accompanying decrease in P4 concentration. qPCR expression of 20alpha-HSD in CL from control and PGF2alpha-treated buffalo cows showed higher expression at 3 and 18 h post treatment, but its specific activity was not altered at different time points post PGF2alpha treatment. The Nur77 expression increased several fold 3 h post PGF2alpha treatment similar to the increased expression observed in the PGF2alpha-treated pseudo pregnant rats which perhaps suggest initiation of activation of apoptotic pathways in response to PGF2alpha treatment. Conclusions: The results taken together suggest that synthesis of P4 appears to be primarily affected by PGF2alpha treatment in buffalo cows in contrast to increased metabolism of P4 in rodents.

Evaluación del sistema de producción de leche "El Corpus" El Menco, Rivas

Para la realización de este estudio se plantearon los siguientes objetivos: contribuir al conocimiento y mejoramiento del sistema de producción de leche "El Corpus" El Menco, Rivas, y de manera específica caracterizar y analizar los componentes del sistema, realizar un anál isis biológico y económico a partir de un estudio de eficiencia y realizar un estudio comparativo de la eficiencia del sistema en dos momentos en el tiempo. Este estudio se realizo en la finca "El Corpus" El Menco, Rivas en el período enero-diciembre del año 2000 y el período julio 2001-junio 2002. La unidad de producción se encuentra localizada a la altura del kilómetro 85 ½ carretera panamericana y 13 kilómetros al este a orilla del lago de Nicaragua, a 70 msnm y una latitud norte de 11°50'16" y una longitud oeste de 85°50'16". Para llevar a cabo la presente investigación se definieron las siguientes etapas generales: Caracterización e identificación del sistema de producción, análisis y comparación del sistema de los dos periodos en estudio, a través de un diagnóstico estático y dinámico. Durante la caracterización se observó que la mayoría de los componentes del sistema presentan óptimas condiciones para lograr una eficiente producción lográndose determinar que existe mal manejo en algunos de ellos. Así mismo en el análisis de eficiencia se logró determinar que las variables productivas del año 2000 en las que se obtuvieron mejores resultados con respecto al período julio 2001-junio 2002 fueron: El porcentaje de parición, porcentaje de mortalidad de terneras y la carga animal, lo cual fue lo contrario para la variable del porcentaje de descarte, la producción de leche/vaca/día y la producción de leche/ha/año. En lo que respecta a las variables reproductivas en las que se obtuvieron mejores resultados fue solamente el numero de servicios por concepción; mientras que para la edad de incorporación, peso de incorporación, edad al primer parto, período de servicio, intervalo entre parto fue lo contrario, no obstante la edad de incorporación y el peso de incorporación, se obtuvieron diferencias significativas a través de la prueba de "t" (a = 0.05); mientras que en la edad al primer parto, período de servicio, intervalo entre parto, no se obtuvieron diferencias. En lo que se refiere a la eficiencia económica del sistema los mejores resultados se obtuvieron en el periodo julio 2001 –junio 2002 el cual fue mas rentable y mas eficiente.

Viel, así en el cielo como en el canon : un lugar en el corpus de la poesía argentina

Resumen: Para analizar cuál es el lugar de Héctor Viel Temperley en las letras argentinas tomamos el movimiento editorial en torno de él, sobre todo la edición y reedición de sus obras y, por otro lado, relevamos antologías poéticas. En la labor, sorpresivamente, aparecen rasgos reveladores que desmienten el supuesto acerca de su haber sido ignoto; por otro lado, hemos hallado elementos que iluminan áreas no estudiadas, las que en nuestra investigación —aún en marcha— se van mostrando fundantes. Asimismo, consideramos su papel como ícono motivador en algunas obras recientes, movilizador, afectiva y conceptualmente, de cierto cúmulo de contenidos.

A methodology for the semiautomatic annotation of EPEC-RolSem, a basque corpus labeled at predicative level following the PropBank-Verb Net model

In this article we describe the methodology developed for the semiautomatic annotation of EPEC-RolSem, a Basque corpus labeled at predicate level following the PropBank-VerbNet model. The methodology presented is the product of detailed theoretical study of the semantic nature of verbs in Basque and of their similarities and differences with verbs in other languages. As part of the proposed methodology, we are creating a Basque lexicon on the PropBank-VerbNet model that we have named the Basque Verb Index (BVI). Our work thus dovetails the general trend toward building lexicons from tagged corpora that is clear in work conducted for other languages. EPEC-RolSem and BVI are two important resources for the computational semantic processing of Basque; as far as the authors are aware, they are also the first resources of their kind developed for Basque. In addition, each entry in BVI is linked to the corresponding verb-entry in well-known resources like PropBank, VerbNet, WordNet, Levin’s Classification and FrameNet. We have also implemented several automatic processes to aid in creating and annotating the BVI, including processes designed to facilitate the task of manual annotation.