Abstract:

Although several reports indicate effects of histamine (HA) on female reproductive functions, scant literature exists to suggest a physiological role of HA in the male gonad. In the present study, we report a dual concentration-dependent effect of HA on steroidogenesis in MA-10 murine Leydig cells and purified rat Leydig cells. Although 1 nM HA can stimulate steroid production and significantly increase the response to LH/hCG in these cells, 10 microM HA exerts an inhibitory effect. We also provide confirming evidence for the existence of functional HRH1 and HRH2 receptors in both experimental models. The use of HRH1 and HRH2 selective agonists and antagonists led us to suggest that HRH2 activation would be largely responsible for stimulation of steroidogenesis, while HRH1 activation is required for inhibition of steroid synthesis. Our results regarding signal transduction pathways associated with these receptors indicate the coupling of HRH2 to the adenylate cyclase system through direct interaction with a Gs protein. Moreover, we show HRH1 activation mediates increases in inositol phosphate production, possibly due to coupling of this receptor to Gq protein and phospholipase C activation. The data compiled in this report clearly indicate that HA can modulate Leydig cell steroidogenesis in the testis and suggest a possible new physiological site of action for HA. Given that many drugs binding to HRH1, HRH2, or both, are widely prescribed for the treatment of diverse HA-related pathologies, it seems necessary to increase the knowledge regarding histaminergic regulation of testicular functions, to avoid possible unexpected side effects of such substances in the testis.

Author affiliation: Mondillo, Carolina. 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; Argentina

Author affiliation: Patrignani, Zoraida. 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; Argentina

Author affiliation: Reche, Cecilia. 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; Argentina

Author affiliation: Rivera, Elena. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina

Author affiliation: Pignataro, Omar Pedro. 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; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina

Abstract:

Estrogens inhibit androgen production and this negative action on amphibian steroidogenesis could be related to the regulation of steroidogenic enzymes. Estrogens are also involved in the regulation of amphibian spermatogenesis by controlling testicular apoptosis and spermatogonial proliferation. The Bidder's organ (BO) is a structure characteristic from the Bufonidae family and in adult males of Rhinella arenarum it is one of the main sources of plasma estradiol (E2). The purpose of this study is to analyze the effect of E2 on testicular steroidogenic enzymes, apoptosis and proliferation in the toad R. arenarum. For this purpose, testicular fragments were treated during 24h with or without 2 or 20nM of E2. After treatments, the activities of cytochrome P450 17α-hydroxylase-C17-20 lyase (CypP450c17) and 3β-hydroxysteroid dehydrogenase/isomerase (3β-HSD/I) were measured by the transformation of radioactive substrates into products, and CypP450c17 expression was determined by Western blot analysis. Apoptosis in testicular sections was detected with a commercial fluorescent kit based on TUNEL method, and proliferation was evaluated by BrdU incorporation. Results indicate that E2 has no effect on CypP450c17 protein levels or enzymatic activity, while it reduces 3β-HSD/I activity during the post reproductive season. Furthermore, although E2 has no effect on apoptosis during the pre and the post reproductive seasons, it stimulates testicular apoptosis during the reproductive season, mostly in spermatocytes. Finally, E2 has no effect on testicular proliferation all year long. Taken together, these results suggest that E2 is involved in the regulation of testicular steroidogenesis and spermatogenesis.

Author affiliation: Scaia, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Endocrinología Comparada; Argentina

Author affiliation: Volonteri, María Clara. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Endocrinología Comparada; Argentina

Author affiliation: Czuchlej, Silvia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Endocrinología Comparada; Argentina

Author affiliation: Ceballos, Nora Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Endocrinología Comparada; Argentina

Abstract:

In the present study, we demonstrate the expression of heme oxygenase (HO) isozymes, HO-1 and HO-2 (listed as HMOX1 and HMOX2 in the MGI Database), in MA-10 Leydig tumor cells and its effect on steroidogenesis. The well-known HO inducer, hemin, increased both HO-1 and HO-2 protein levels and HO-specific activity. Induction of HO by hemin inhibited basal, hCG-, and dibutyryl cAMP (db-cAMP)-induced steroidogenesis in a reversible way. When we studied the effect of HO isozymes along the steroid synthesis, we found that steroidogenic acute regulatory protein levels were decreased, and the conversion of cholesterol to pregnenolone was inhibited by hemin treatment, with no changes in the content of cholesterol side-chain cleavage enzyme (P450scc). hCG and db-cAMP also stimulated the expression of HO-1 and HO-2, and HO enzymatic activity in MA-10 cells. Basal and hCG-stimulated testosterone synthesis was also inhibited by hemin in rat normal Leydig cells. Taken together, these results suggest that: i) at least one of HO products (presumably carbon monoxide) inhibits cholesterol transport to the inner mitochondrial membrane and Leydig cell steroidogenesis by binding to the heme group of the cytochrome P450 enzymes, in a similar way as we described for nitric oxide, and ii) hCG stimulation results in the induction of an antioxidant enzymatic system (HO) acting as a cytoprotective mechanism in Leydig cells, as already demonstrated in the adrenal gland.

Author affiliation: Piotrkowski, Barbara. 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; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina

Author affiliation: Monzón, Casandra Margarita. 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; Argentina

Author affiliation: Pagotto, Romina María del Luján. 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; Argentina

Author affiliation: Reche, Cecilia. 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; Argentina

Author affiliation: Besio Moreno, Marcos Alberto. 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; Argentina

Author affiliation: Cymeryng, Cora Betriz. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Author affiliation: Pignataro, Omar Pedro. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Bioquímica Humana; Argentina. 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; Argentina

Abstract:

Rhinella arenarum is a South American toad with wide geographic distribution. Testes of this toad produce high amount of androgens during the non reproductive season and shift steroid synthesis from androgens to 5α-pregnanedione during the breeding. In addition, plasma estradiol (E 2 ) in males of this species shows seasonal variations but, since testes of R. arenarum do not express aromatase, the source of plasma E 2 remained unknown for several years. However, the Bidder's organ (BO), a structure located at one pole of each testis, is proposed to be the main source of E 2 in male's toads since it expresses several steroidogenic enzymes and is able to produce E 2 from endogenous substrates throughout the year. In addition, there were significant correlations between plasma E 2 and total activity of BO aromatase, and between plasma E 2 and the amount of hormone produced by the BO in vitro. In the toad, apoptosis induced by in vitro treatment with E 2 was mostly detected in spermatocytes during the breeding and in spermatids during the post-reproductive season, suggesting that this steroid has an important role in controlling spermatogenesis. However, in vitro treatment with E 2 had no effect on proliferation. This evidence suggests that the mechanism of action of E 2 on amphibian spermatogenesis is complex and more studies are necessary to fully understand the role of estrogens regulating the balance between cellular proliferation and apoptosis. In addition, in R. arenarum in vitro studies suggested that E 2 has no effect on CypP450c17 protein levels or enzymatic activity, while it reduces 3β-hydroxysteroid dehydrogenase/isomerase (3β-HSD/I) activity during the post reproductive season. As well, E 2 regulates FSHβ mRNA expression all over the year suggesting a down regulation process carried out by this steroid. The effect on LHβ mRNA is dual, since during the reproductive season estradiol increases the expression of LHβ mRNA while in the non-reproductive season it has no effect. In conclusion, the effect of E 2 on gonadotropins and testicular function is complex, not clearly understood and probably varies depending on the species. The aim of the current article is to review evidence on reproductive endocrinology and on the role of estradiol regulating reproduction in amphibians, with emphasis on the South American species Rhinella arenarum.

Author affiliation: Scaia, María Florencia. 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

Author affiliation: Volonteri, María Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Diversidad y Evolución Austral; Argentina

Author affiliation: Czuchlej, Silvia Cristina. Universidad de Buenos Aires; Argentina

Author affiliation: Ceballos, Nora Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina

Abstract:

Hormonal regulation of steroidogenesis involves arachidonic acid (AA) metabolism through the 5-lipoxygenase pathway. One of the products, 5-hydroperoxy-eicosatetraenoic acid (5-HpETE), acts as a modulator of the activity of the steroidogenic acute regulatory (StAR) protein promoter. Besides, an oxoeicosanoid receptor of the leukotriene receptor family named OXE-R is a membrane protein with high affinity and response to 5-HpETE, among other AA derivatives. The aim of our work was to elucidate whether this receptor may be involved in steroidogenesis. RT-PCR and western blot analysis demonstrated the presence of the mRNA and protein of the receptor in human H295R adrenocortical cells. The treatment of H295R or MA-10 cells (murine Leydig cell line) with 8Br-cAMP together with docosahexaenoic acid (DHA, an antagonist of the receptor) partially reduced StAR induction and steroidogenesis. On the contrary, 5-oxo-ETE -the prototypical agonist, with higher affinity and potency on the receptor- increased cAMP-dependent steroid production, StAR mRNA and protein levels. These results lead us to conclude that AA might modulate StAR induction and steroidogenesis, at least in part, through 5-HpETE production and activation of a membrane receptor, such as the OXE-R

Author affiliation: Cooke, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina

Author affiliation: Di Consoli, Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina

Author affiliation: Maloberti, Paula Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina

Author affiliation: Cornejo Maciel, Maria Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina

Abstract:

Histamine (HA) is a neurotransmitter synthesized in most mammalian tissues exclusively by histidine decarboxylase enzyme. Among the plethora of actions mediated by HA, the modulatory effects on steroidogenesis and proliferation in Leydig cells (LCs) have been described recently. To determine whether the effects on LCs reported could be extrapolated to all steroidogenic systems, in this study, we assessed the effect of this amine on adrenal proliferation and steroidogenesis, using two adrenocortical cell lines as experimental models, murine Y1 cells and human NCI-H295R cells. Even when steroidogenesis was not modified by HA in adrenocortical cells, the biogenic amine inhibited the proliferation of H295R cells. This action was mediated by the activation of HRH1 subtype and an increase in the production of inositol phosphates as second messengers, causing cell-cycle arrest in the G2/M phase. These results indicate a new role for HA in the proliferation of human adrenocortical cells that could contribute to a better understanding of tumor pathology as well as to the development of new therapeutic agents.

Author affiliation: Pagotto, Romina María del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina

Author affiliation: Pereyra, Elba Nora. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina

Author affiliation: Monzón, Casandra Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina

Author affiliation: Mondillo, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina

Author affiliation: Pignataro, Omar Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina

Abstract:

Cystic ovarian disease (COD) represents an important cause of infertility in dairy cattle and is associated with multiple physiological disorders. Steroidogenesis, which is necessary to ensure normal ovarian functions, involves multiple enzymatic pathways coordinated by insulin and other proteins. We have previously shown that cows with COD have an altered insulin response. Therefore, in the present study, we evaluated further alterations in intermediates downstream of the PI3K pathway and pathways mediated by ERK as critical signals for the expression of steroidogenic enzymes in the ovaries of control cows and cows with spontaneous COD. To this end, we evaluated the gene and protein expression of pan-AKT, mTOR, ERK1/2, and steroidogenic enzymes by real-time PCR and immunohistochemistry. Steroid hormone concentrations were assessed at systemic and intrafollicular level. Results showed altered expression of intermediate molecules of the insulin signaling pathway, whose action might modify the synthetic pathway of steroidogenic hormones. Similarly, the expression of steroidogenic enzymes and the concentration of progesterone in serum and follicular fluid were altered. These alterations support the hypothesis that systemic factors contribute to the development and/or maintenance of COD, and that metabolic hormones within follicles such as insulin exert determinant effects on ovarian functionality in cows with COD.

Author affiliation: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Huber, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Rodríguez, Fernanda Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Author affiliation: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

Abstract:

La FSH se sintetiza y secreta como una familia de variantes glicosiladas, las cuales difieren en el grado de sialilación y complejidad de sus oligosacáridos y en la inducción de respuestas biológicas en células blanco, in vivo e in vitro. El objetivo general de esta Tesis Doctoral fue establecer la relación existente entre las características de los carbohidratos presentes en la molécula de FSH y la inducción de respuestas biológicas en las células de la granulosa. La acción de variantes glicosiladas, aisladas de FSH recombinante humana (FSHrh), fue evaluada in vitro utilizando dos modelos experimentales. Se estudió la producción de inhibinas diméricas en células de la granulosa de rata; de esteroides e inhibinas en la línea celular de granulosa humana, KGN, como así también el patrón de expresión génica global y la expresión de enzimas esteroidogénicas y subunidades de inhibinas. Estudios in vitro demostraron que el grado de sialilación y la complejidad de los oligosacáridos presentes en la FSHrh reguló en forma diferencial la producción de esteroides y péptidos. Asimismo, el análisis de ontología génica reveló la capacidad que poseen las variantes glicosiladas de FSHrh de modular en forma diferencial la expresión de genes involucrados en funciones o procesos esenciales para el mantenimiento de la función celular. Las características de las variantes glicosiladas de la FSH circulante fueron determinadas en condiciones fisiológicas y en pacientes infértiles con ciclos regulares. Se observaron alteraciones en el grado de sialilación de la FSH circulante en las pacientes con trastornos reproductivos y en mujeres jóvenes ovodonantes. Los resultados obtenidos demuestran que los oligosacáridos de la molécula de FSH regulan distintas funciones de la célula de la granulosa. Alteraciones precoces en el grado de sialilación de la gonadotrofina podrían comprometer la capacidad reproductiva en la mujer.

FSH is produced and secreted as a family of glycosylation variants which differ from each other in the oligosaccharide sialylation and complexity and in the induction of different biological responses in vitro and in vivo. The aim of this Doctoral Thesis was to establish the relationship between the FSH carbohydrate characteristics and the induction of biological responses in granulosa cells. The effect of recombinant human FSH glycosylation variants were evaluated in two experimental models in vitro. Dimeric inhibin production was assessed in rat granulosa cells, steroids and inhibin production in human granulosa cell line KGN, as well as global gene expression profile and the expression of steroidogenic enzymes and inhibin subunits. In vitro studies demonstrated that sialylation and complexity of rhFSH oligosaccharides induced a differential regulation of steroid and inhibin production. Gene ontology analysis revealed the ability of rhFSH glycosylation variants to differentially modulate the expression of genes involved in essential cell biological processes and molecular functions. Circulating FSH oligosaccharides characteristics were determined under physiological conditions and in infertile patients. Alterations in FSH sialylation were observed in infertile patients and in ovodonors. The results obtained in this study show that FSH oligosaccharides regulate different functions in granulosa cells. Early alterations in FSH sialylation may jeopardize the reproductive capacity in women.

Author affiliation: Loreti, Rosana Nazareth. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.

Abstract:

La histamina (HA) es una amina biógena asociada a diversas funciones llevadas a cabo a través de cuatro subtipos de receptores pertenecientes a la familia de los GPCR. En este trabajo de Tesis se evaluó el efecto de HA sobre la esteroidogénesis y proliferación de tumores córticoadrenales utilizando dos líneas celulares, las Y1 murinas y las H295R humanas y se profundizaron estudios previos del laboratorio en células de Leydig (CL), analizando la participación de esta amina en su desarrollo. En las células córticoadrenales la esteroidogénisis no se vió modificada por HA, lo que supone que el efecto modulador reportado previamente en CL no sería extrapolable a todos los sistemas esteroidogénicos. En cambio, HA inhibió la proliferación de las células H295R a 24h, provocando un arresto de las células en la fase G2/M en un mecanismo mediado por el subtipo de receptor H1 e IP3 y DAG como segundos mensajeros. Estos resultados permitieron identificar un nuevo rol de HA sobre la proliferación de células córticoadrenales humanas que podría contribuir a una mejor comprensión de la patología tumoral y al desarrollo de nuevos agentes terapéuticos. En lo que respecta a las CL, HA fue capaz de inhibir la proliferación de CL inmaduras (CLI) en ensayos in vitro a través de la activación del subtipo de receptor H4, mientras que en estudios in vivo el tratamiento con esta amina produjo un incremento en el número de CL con morfología adulta. Estos datos sugieren que esta amina modularía la proliferación en un paso previo a la diferenciación y pone en evidencia la necesidad de continuar los estudios sobre su función en la transición de los distintos estadíos de desarrollo de CL así como en la génesis de patologías tumorales.

Histamine (HA) is a biogenic amine associated with heterogeneous functions carried out through the activation of four receptor subtypes, all of them members of the GPCR family. In this doctoral thesis the effects of HA on steroidogenesis and proliferation of adrenocortical tumors were evaluated using two experimental models, the murine Y1 and the human H295R cell lines. Also previous studies in Leydig cells (LC) were continued, evaluating the participation of the cited amine in their development. Steroidogenesis was not modified by HA in adrenocortical cells, which suggests that the modulatory effect previously reported in LC can not be extrapolated to all steroidogenic systems. Nevertheless, HA inhibited the proliferation of H295R cells after a 24 h-treatment through activation of the H1 receptor subtype and an increase in the production of IP3 and DAG second messengers, causing a cell cycle arrest in the G2/M phase. These results indicate a new role of HA on human adrenocortical cells proliferation that could contribute to a better understanding of tumor pathology, and to the development of new therapeutic agents. With respect to LC, HA was able to inhibit the proliferation of immature and progenitor LC in in vitro assays via the activation of the H4 receptor subtype, whereas in in vivo studies HA caused an increase in LC number. These data suggest that the amine would modulate the proliferation of LC before differentiation takes place and highlights the need for further studies on its role on the transition of different LC development stages as well as on its influence on tumor pathogenesis.

Author affiliation: Pagotto, Romina María del Luján. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.

Abstract:

La melatonina, hormona secretada por la glándula pineal, modula los ritmos biológicos diarios y los ciclos de sueño/vigilia. A través de su acción regulatoria sobre la producción y secreción de hormonas hipotalámicas e hipofisarias, participa también en la modulación de la actividad sexual en especies con ciclos reproductivos anuales controlados por el fotoperíodo. En este Trabajo de Tesis Doctoral se investigaron dos mecanismos de acción diferenciales de la melatonina en el testículo: su rol como hormona y su rol como agente anti/pro oxidante. Los estudios realizados permitieron establecer: a) las señales intracelulares asociadas al rol inhibitorio ejercido por el sistema melatonina/CRH sobre la regulación de la esteroidogénesis en células de Leydig del hámster Dorado, b) la concentración testicular de melatonina y la expresión de los principales componentes de la cascada de señalización del sistema melatonina/CRH en el testículo de pacientes que padecen infertilidad idiopática, y c) la acción regulatoria ejercida por melatonina sobre la proliferación, estado oxidativo y capacidad secretoria en macrófagos y mastocitos testiculares. La melatonina desempeñaría un rol clave en la modulación de la esteroidogénesis testicular en el hámster. En el testículo humano patológico, ejercería además acciones regulatorias sobre la población local de macrófagos y mastocitos.

Melatonin is a hormone secreted by the pineal gland that mediates the physiological regulation of both daily rhythms and sleeping/waking cycles. In seasonal breeders, this hormone also participates in the modulation of annual reproductive activity through its action on the hypothalamic-pituitary axis. In this Doctoral Thesis two differential mechanisms of action of melatonin in the testis were studied: its role as a hormone and its role as an anti/pro-oxidant agent. Studies performed in this Thesis allowed us to establish: a) the intracellular events triggered by melatonin/CRH that lead to a down-regulation of the steroidogenic process in Syrian hamster Leydig cells, b) the testicular melatonin concentration and the expression of the main components of the signaling cascade linked to the melatonin/CRH system in testicular biopsies of men suffering from idiopathic infertility, c) the effects of melatonin on proliferation, oxidative state and secretory capability in testicular macrophages and mast cells. Thus, melatonin would play a key role in the regulation of testicular steroidogenesis in the hamster. In the pathological human testis, melatonin would also participate in the modulation of the local macrophage and mast cell populations.

Author affiliation: Rossi, Soledad Paola. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.