{"id":116578,"date":"2014-03-11T00:00:00","date_gmt":"2014-03-11T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/estudio-de-la-funcion-de-la-hormona-estimuladora-del-fola%c2%adculo-de-lubina-su-implicacion-en-la-espermatogenesis-y-sus-rutas-de-senalizacion-intracelular\/"},"modified":"2014-03-11T00:00:00","modified_gmt":"2014-03-11T00:00:00","slug":"estudio-de-la-funcion-de-la-hormona-estimuladora-del-fola%c2%adculo-de-lubina-su-implicacion-en-la-espermatogenesis-y-sus-rutas-de-senalizacion-intracelular","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/fisiologia-animal\/estudio-de-la-funcion-de-la-hormona-estimuladora-del-fola%c2%adculo-de-lubina-su-implicacion-en-la-espermatogenesis-y-sus-rutas-de-senalizacion-intracelular\/","title":{"rendered":"Estudio de la funci\u00f3n de la hormona estimuladora del fol\u00edculo de lubina: su implicaci\u00f3n en la espermatog\u00e9nesis y sus rutas de se\u00f1alizaci\u00f3n intracelular."},"content":{"rendered":"<h2>Tesis doctoral de <strong> Mar\u00eda  Jose Mazon Moya <\/strong><\/h2>\n<p>Resumen  el proceso reproductivo en peces, al igual que en el resto de vertebrados, est\u00e1 regulado por una cascada hormonal en la que est\u00e1n implicados el cerebro, la hip\u00f3fisis y las g\u00f3nadas, conocido como eje chg. Dos de las principales hormonas implicadas en el control de la reproducci\u00f3n son las gonadotrofinas fsh (hormona estimuladora del fol\u00edculo) y lh (hormona luteinizante), ambas producidas en la hip\u00f3fisis y liberadas tras el est\u00edmulo con las hormonas liberadoras de gonadotrofinas (gnrh) y con acci\u00f3n a nivel gonadal.  desde la perspectiva de la producci\u00f3n animal en general, y de la acuicultura en particular, es esencial conocer en la especie a domesticar los mecanismos b\u00e1sicos de control endocrino. Este conocimiento b\u00e1sico es de gran utilidad para poder manipular la reproducci\u00f3n a conveniencia y al mismo tiempo poder desarrollar estrategias terap\u00e9uticas para solventar problemas de fertilidad que en ocasiones presentan los stocks de reproductores. Sin embargo, como consecuencia de la limitaci\u00f3n en la disponibilidad de herramientas para su estudio, para una gran mayor\u00eda de las especies cultivables de peces se desconocen los mecanismos particulares que controlan su reproducci\u00f3n. en esta tesis se optimiza una alternativa nueva de administraci\u00f3n de gonadotrofinas recombinantes, la tecnolog\u00eda conocida como transferencia g\u00e9nica som\u00e1tica. En los cap\u00edtulos 3 y 4 se inyectaron adultos y juveniles de lubina con construcciones de dna codificante de las gonadotrofinas de cadena \u00fanica lh y fsh, respectivamente, y se evalu\u00f3 el incremento de estas hormonas en el torrente sangu\u00edneo. En ambos experimentos las inyecciones de dna se compararon con la inyecci\u00f3n directa de la gonadotrofina recombinante correspondiente. Estos cap\u00edtulos muestran que las construcciones de dna se incorporan a las c\u00e9lulas musculares en el lugar de inyecci\u00f3n, y las hormonas codificadas son sintetizadas y liberadas al torrente sangu\u00edneo. los cap\u00edtulos 3 y 4 confirman que la transferencia g\u00e9nica som\u00e1tica es una herramienta poderosa para realizar estudios funcionales in vivo con gonadotrofinas, siendo adem\u00e1s \u00fatil como terapia hormonal en peces, present\u00e1ndose como alternativa a los tratamientos tradicionales con hcg o gnrh. la funcionalidad de las hormonas recombinantes sintetizadas por las c\u00e9lulas musculares de lubina tras la incorporaci\u00f3n del dna, se determin\u00f3 estudiando su efecto en el desarrollo de los test\u00edculos. En los cap\u00edtulos 4 y 5 se evalu\u00f3 como la acci\u00f3n de la fsh es esencial para iniciar el proceso de espermatog\u00e9nesis, promoviendo la proliferaci\u00f3n de las espermatogonias, siendo necesaria la acci\u00f3n posterior de la lh para culminar la espermiaci\u00f3n. Los individuos inyectados s\u00f3lo con el pl\u00e1smido de fsh presentaron una espermatog\u00e9nesis parcial, sin indicios de espermiaci\u00f3n, estado que s\u00f3lo alcanzaron aquellos animales que se inyectaron de manera secuencial con los pl\u00e1smidos de fsh y lh. Estos experimentos tambi\u00e9n demuestran que la lh por si sola no es capaz de promover la maduraci\u00f3n testicular. Adem\u00e1s en los an\u00e1lisis de expresi\u00f3n g\u00e9nica se observ\u00f3 que los tratamientos con fsh incrementaron la expresi\u00f3n de marcadores de c\u00e9lulas germinales como piwi, pcna y scp3, mientras que el efecto de la lh se detect\u00f3 en enzimas esteroidog\u00e9nicas como cyp17a1 y cyp17a2. en esta tesis tambi\u00e9n se describe c\u00f3mo una de las acciones m\u00e1s directas de la fsh es la estimulaci\u00f3n de la producci\u00f3n de 11kt. Con el fin de analizar m\u00e1s en detalle la acci\u00f3n espec\u00edfica de la fsh en la activaci\u00f3n de la esteroidog\u00e9nesis, y de definir mecanismos de regulaci\u00f3n conservados entre diferentes especies de peces, se estudi\u00f3 in vitro la acci\u00f3n de la fsh en cultivo primario de test\u00edculo y ovario de lubina y pez cebra.  en el cap\u00edtulo 6 se describe c\u00f3mo la fsh emplea principalmente la ruta del ampc\/pka para transmitir su se\u00f1al una vez activado su receptor, no siendo \u00e9sta la \u00fanica v\u00eda, ya que tambi\u00e9n es importante la actividad de las mapk. Haciendo uso de inhibidores y estimuladores de ambas rutas se ha perfilado un escenario en el que la fsh utiliza de manera independiente las rutas del ampc\/pka o las mapk para estimular la transcripci\u00f3n de genes implicados en la esteroidog\u00e9nesis, como star o  cyp19a1a   abstract the reproductive process in fish, as in all vertebrate species, is tightly regulated by a hormonal axis, called bpg, involving the brain, pituitary and gonads. Two major hormones that control reproduction are the gonadotropins fsh (follicle-stimulating hormone) and lh (luteinizing hormone) are both produced at the pituitary level, released to the bloodstream after the stimulus of the gnrh (gonadotropin release hormone), and with action at  the gonad level.  from a fish farming perspective, it is essential to fully understand the endocrine mechanism underlying reproductive performance. This knowledge is important to manipulate reproductive behaviour and for the development of therapeutic strategies aiming to solve fertility issues frequently present in the broodstocks. However, due to the limited tools available for the vast majority of the cultivable species, the mechanisms controlling their reproduction are not well characterized. this thesis describes an alternative administration of recombinant gonadotropins using the technique of somatic gene transfer. In chapters 3 and 4, adult and juvenile sea bass were injected with a plasmid dna encoding lh and fsh gonadotropin sequences in a single chain form, and the levels of either  gths were analysed in the bloodstream. In both experiments the plasma levels of gths attained by plasmid dna injected fish were compared with plasma levels of fish injected with the equivalent recombinant gth. Both chapters clearly show that plasmid dna was efficiently incorporated by the cell muscle, and the encoded hormones were synthetized and released to the bloodstream. These results confirm that somatic gene transfer can be a powerful tool for in vivo functional studies of gonadotropin. In addition, this technique can be used for hormone therapy in fish, and can offer a useful alternative to classic treatments with hcg or gnrh. the functionality of the recombinant hormones synthetized by cell muscles after plasmid dna injection was evaluated by studying the development of testes. Chapters 4 and 5 describe the crucial role of fsh in starting the process of spermatogenesis, promoting the proliferation of spermatogonia cells, and its requirements for lh to reach the spermiation process.  Fish receiving only fsh plasmid injections showed partial spermatogenesis, with no evidence for spermiation. Only the fish injected with fsh and lh plasmid sequentially reached this late step of the spermatogenesis process. Fish receiving lh plasmid alone showed that gth cannot trigger testis maturation alone. Furthermore, gene expression analysis highlighted the action of fsh on promoting cell proliferation by increasing the expression of germ cell markers such as piwi, pcna piwi, pcna and scp3. By contrast, the effect of lh was more obvious in transcript levels of steroidogenic enzymes such as cyp17a1 and cyp17a2. this thesis additionally describes a direct action of fsh on the stimulation of 11kt production. In order to analyse the specific activity of fsh on the steroidogenesis process in detail, and to define regulatory mechanisms conserved across fish species, the action of fsh in vitro was studied using primary gonad tissue culture from sea bass and zebrafish. Chapter 6 describes the use of the signalling pathway camp\/pka by fsh to transduce the signal after the activation of its receptor not clear. Importantly, camp\/pka is not the only pathway used by fsh, the mapk route is also involved in fsh signal transduction. The use of camp\/pka and mapk inhibitors and stimulators suggest the possibility that fsh can use both routes independently to activate the transcription of some steroidogenic genes such as star and cyp19a1a<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Estudio de la funci\u00f3n de la hormona estimuladora del fol\u00edculo de lubina: su implicaci\u00f3n en la espermatog\u00e9nesis y sus rutas de se\u00f1alizaci\u00f3n intracelular.<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Estudio de la funci\u00f3n de la hormona estimuladora del fol\u00edculo de lubina: su implicaci\u00f3n en la espermatog\u00e9nesis y sus rutas de se\u00f1alizaci\u00f3n intracelular. <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Mar\u00eda  Jose Mazon Moya <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Murcia<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 03\/11\/2014<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Silvia Zanuy Doste<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: francesc Piferrer circuns <\/li>\n<li>mercedes Bl\u00e1nquez peinado (vocal)<\/li>\n<li>Elena Chaves pozo (vocal)<\/li>\n<li>Francisco Prat baella (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Mar\u00eda Jose Mazon Moya Resumen el proceso reproductivo en peces, al igual que en el resto de 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