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Identification of Conserved Genes Triggering Puberty in European Sea Bass Males ( Dicentrarchus Labrax ) by Microarray Expression Profiling

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Identification of Conserved Genes Triggering Puberty in European Sea Bass Males ( Dicentrarchus Labrax ) by Microarray Expression Profiling Retinoic acid signaling pathway: gene regulation during the onset of puberty in the European sea bass Ruta de señalización del ácido retinoico: regulación génica durante el inicio de la pubertad en la lubina Europea Paula Javiera Medina Henríquez Thesis presented to obtain the Ph.D. degree at the Polytechnic University of Catalonia (UPC) Ph. D. Program in Marine Sciences 2019 Thesis supervisor: Dra. Mercedes Blázquez Peinado (ICM-CSIC) Barcelona, 2019 Esta Tesis ha sido realizada en el Departamento de Recursos Marinos Renovables del Instituto de Ciencias del Mar de Barcelona (ICM-CSIC). Parte de la experimentación también se ha llevado a cabo en el Instituto de Acuicultura Torre de la sal (IATS-CSIC), a través de una estrecha colaboración con el grupo de Fisiología de la Reproducción de Peces. La financiación económica para la e xperimentación se ha recibido a través de los siguientes Proyectos de Investigación: Mejora de la producción en acuicultura mediante el uso de herramientas biotecnológicas CSD2007-00002 (AQUAGENOMICS). Bases moleculares, celulares y endocrinas de la pubertad y del desarrollo gonadal en lubina ( Dicentrarchus labrax ). Desarrollo de tecnologías ambientales, hormonales, y de manipulación genética para su control PROMETEO/2010/003 (REPROBASS). Regulación Hormonal y desarrollo del eje hipotalámico-hipofisario-gonadal durante la diferenciación sexual y la pubertad en peces teleósteos PIE 200930I037(TERMOBASS). Regulación endocrina y paracrina de la diferenciación sexual y el desarrollo gonadal en la lubina AGL2011-28890 (REPROSEX). Acción complementaria al proyecto: regulación endocrina y paracrina de la diferenciación sexual y desarrollo gonadal en la lubina (REPROSEX). Avances en el control de la pubertad y ciclo reproductor de la lubina y su aplicación tecnológica a otras especies de peces PROMETEO II/2014/051 (REPOBASS II). Mecanismos endocrinos y paracrinos en el control de la diferenciación y desarrollo de las células germinales de peces AGL2015-67 477-C2-2-R (PUBERGAMETE). Paula Medina fue auspiciada por la beca de doctorado para académicos en el extranjero, proyecto ANT0806 de la Universidad de Antofagasta, Chile. Los resultados de esta tesis han sido presentados en los siguientes congresos: 10º Congreso de la Asociación Ibérica de Endocrinología Comparada. Castellón, España. 23-25 de Septiembre, 2015. “Characterization of aldehyde dehydrogenase family 1a in Dicentrarchus labrax ”. Presentación poster. 27th Conference of European Comparative Endocrinologists, Rennes, France, 25-29th August, 2014. “Retinoic acid signaling pathway during sex differentiation and puberty in the European sea bass ( Dicentrarchus labrax )”. Presentación Oral. 11th International Congress on the Biology of Fish. Edinburgh, Scotland. August 3- 7th, 2014. “Drospirenone effects in steroid levels in blood and hepatic metabolism in European sea bass ( Dicentrarchus labrax )”. Presentación Poster. 17th International Congress of Comparative Endocrinology, Barcelona, Spain. July 15- 19th, 2013. “Transcriptomic changes during the onset of spermatogenesis in the European sea bass”. Presentación Oral. Barcelona, 2019 9th Congreso de la Asociación Ibérica de Endocrinología Comparada, Barcelona, Spain. July 13-14th, 2013. “Testicular organ culture for the study of sea bass spermatogenesis”. Presentación poster. Premio a la mejor presentación. Los resultados de esta tesis forman parte de las siguientes publicaciones: Medina, P., Gómez, A., Zanuy, S., and Blázquez, M. (2019). Involvement of the retinoic acid signaling pathway in sex differentiation and pubertal development in the European sea bass Dicentrarchus labrax . Heliyon 5, e01201. Blázquez, M., Medina, P., Crespo, B., Gómez, A., and Zanuy, S. (2017). Identification of conserved genes triggering puberty in European sea bass males ( Dicentrarchus labrax ) by microarray expression profiling. BMC Genomics 18, 441. Blázquez, M., Crespo, B., Medina, P., Gómez, A. y Zanuy, S. (2011). Bases moleculares del inicio de la pubertad en machos de lubina. Libro de Actas del XIII Congreso Nacional de Acuicultura. El equilibrio con el Medio Ambiente. (M. Villaroel, L. Reig, C. Almansa, eds.) ISBN 978-84-937611-9-6, pp. 57-58. Medina, P., Gómez, A., Zanuy, S., Bláquez, M. (2014). Testicular organ culture for the study of sea bass spermatogenesis. Advances in Comparative Endocrinology, vol VII. Public. Univ. Barcelona (I. Navarro, J. Gutiérrez y E. Capilla, eds.) ISBN: 974-84-475- 3851-5, pp. 145-148. Otras publicaciones con metodologías empleadas en esta tesis y realizadas durante el periodo de formación doctoral pero que no forman parte directa de los resultados de la misma: Blanco, M., Fernandes, D., Medina, P., Blázquez, M., and Porte, C. (2016). Drospirenone intake alters plasmatic steroid levels and cyp17a1 expression in gonads of juvenile sea bass. Environmental Pollution 213, 541-548. Barcelona, 2019 A mi hijo, Vicente Antai Padilla Medina If not stated otherwise all illustrations and photographs from the author´s. Cover artwork by Alfonso Medina Bacarrezza. Graphic design: Tanya Riquelme ([email protected]). Barcelona, 2019 Agradecimientos. En un viaje largo no podemos decidir lo que nos va a suceder, pero si podemos decidir qué hacer con el tiempo que se nos ha concedido. Primero quisiera agradecer a mi maestro ya fallecido, mi profesor de zoología y colega don Ismael Kong Urbina él siempre dijo que si hay alguien que pueda terminar una tesis doctoral, esa eres tú. No solo el creyó en mi, sino también una pareja de grandes científicos de Torre de la Sal. Sin conocerme Silvia Zanuy y Manuel Carrillo tomaron un voto de confianza y me brindaron todo su apoyo para ser parte de su equipo de investigación. Este gesto cambió mi vida, por eso siempre los recordaré y les estaré eternamente agradecida. A mí querida directora de tesis Mercedes Blázquez. Gracias por ser una de mis maestras de vida, gracias por tener oído, paciencia y empatía con todas las historias del mundo y siempre, siempre, siempre, esa palabra sanadora que te empuja hacia adelante para entregar lo mejor de ti y crecer, tal y como lo hace ella. Han sido años de trabajo y amistad, gracias por esperarme en la orilla, al fin llegué!!!. Simplemente gracias para siempre. A mis compañeros de viaje, confidentes, aliados y amigos del IATS. No podría haber completado ni la mitad de mis propósitos sin Berta, Majo, mi archienemigo Olivier, Vicky, Felipe, Vanesa y Raúl Cortés, por mi parte nuestra amistad trasciende la distancia y el tiempo, siempre los llevaré en mi corazón. Especialmente a mi compatriota el Sebas, su ayuda fue fundamental en muchos momentos, incluso antes de conocernos y lo sigue siendo, gracias chango. Gracias a Silvia, Gemma y Elvira. Gracias por su trabajo diario en el laboratorio y la ZAE, por su paciencia conmigo y las lubinas. Gracias chicas por un trabajo impecable!!!. Gracias a todas las lubinas del mundo por ser como son. Agradecer también la ayuda, apoyo y amistad incondicional de mis queridos amigos Clara, Javier, María José, Irene, y Azahar. Su amistad fue fundamental a mi llegada a Valencia y en la distancia durante todo el tiempo que estuve en Castellón y en Barcelona. Representan a mi familia en España. Siempre tendrán un cariño inmenso y un lugar especial en mi corazón. Gracias a las maravillosas voces de la Coral del mar, los Ovnisingers y, mis amigos B&R: Paco, Joan y Angels, y a mis compañeros del ICM: Natalia, Sara y Fran. Gracias a ustedes mi viaje transcurrió como un sueño hermoso. Barcelona, 2019 A mí querida guía y alma amiga, Elena. Gracias amiguín por ayudarme a levantarme cuando estaba en el suelo, por ser brutalmente honesta conmigo y sobre todo gracias por seguir compartiendo la vida conmigo. Nos convertimos en hermanas del alma y nuestras vidas estarán ligadas a pesar del tiempo y la distancia, gracias amiga para siempre. Gracias a mis padres Marta y Alfonso por entregarme siempre la mejor versión de ustedes mismos, por enseñarme a luchar con la cabeza en alto, por amarme y cuidarme incondicionalmente a pesar de mi y de ustedes. Gracias por ser y estar. Gracias a mi hermano, Diego y a su hermosa familia, Francia, Antonella y Alfonsito, por tanto amor y refugio incondicional. Gracias a mi abuelita Kika, por cuidarme siempre en el más acá y en el más allá (bien habilosita). Gracias a mis amigas Denisse, Karla, Viviana y Cynthia por siempre estar ahí, creer en mí y darme ánimos. Gracias a mi compañero Rodrigo. Gracias por la ternura de tu fuerza, por todo tu apoyo y amor. Gracias por no permitir que me olvide quién soy y acompañarme en esta aventura. Finalmente gracias también a Vicente. La vida cantó y dijo tu nombre, eres la luz de mi vida, mi mejor tesis doctoral y por ti soy capaz de todo. La fortaleza de cada uno de ustedes me acompaña día a día, su energía es mi energía y mi continuidad. Así es, fui capaz de completar esta tarea para lograr ver en mi lo que siempre estuvo allí, para permitir que me ayuden y que me amen, para explotar mis talentos y ser reflejo de los que me rodean. Ahora que esta tesis está terminada el verdadero viaje comienza, no sé donde me llevará, pero seguro será épico. Barcelona, 2019 Abbreviation list. 11-KT: 11-Ketotestosterone 4-OH-RA: 4-Hydroxy retinoic acid 4-oxo-RA 4-oxo-Retinoic acid 9cis-RA: 9cis retinoic acid ADH: Alcohol- dehydrogenase AGN193109: Retinoid analog ALDH: Aldehyde-dehydrogenase APROMAR: Asociación Empresarial de Acuicultura de España atRA: All-trans retinoic acid BCMO1: Beta-Carotene 15,15'-MonoOxygenase BMS-204493:
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