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Del Mejillón Choromytilus Chorus (Molina UNIVERSIDAD DE ANTOFAGASTA FACULTAD DE RECURSOS DEL MAR PROGRAMA DE DOCTORADO EN CIENCIAS APLICADAS, MENCIÓN SISTEMAS MARINOS COSTEROS FUNCIÓN DE LOS ESTEROIDES SEXUALES EN LA DIFERENCIACIÓN SEXUAL (GONADAL) DEL MEJILLÓN CHOROMYTILUS CHORUS (MOLINA 1782) (BIVALVIA MYTILIDAE) Tesis presentada a la Facultad de Recursos del Mar como requisito para optar al grado académico de Doctor en Ciencias Aplicadas, mención Sistemas Marinos Costeros MARYORI IVONNE RUIZ VELÁSQUEZ Profesor Patrocinante: Dr. Manuel Zapata Arcos Profesor Copatrocinante: Dra. María Teresa González Enero, 2017 Antofagasta, Chile CALIFICACIONES FACULTAD DE RECURSOS DEL MAR PROGRAMA DE DOCTORADO EN CIENCIAS APLICADAS, MENCIÓN SISTEMAS MARINOS COSTEROS FUNCIÓN DE LOS ESTEROIDES SEXUALES EN LA DIFERENCIACIÓN SEXUAL (GONADAL) DEL MEJILLÓN CHOROMYTILUS CHORUS (MOLINA 1782) (BIVALVIA MYTILIDAE) Dr. Manuel Zapata Arcos ----------------------------------------- Profesor Patrocinante Dra. María Teresa González ----------------------------------------- Profesor Co Patrocinante Dr. Eduardo Tarifeño Silva ----------------------------------------- Profesor Evaluador Externo Dr. Aldo Pacheco Velásquez ----------------------------------------- Profesor Evaluador Programa Enero, 2017 Antofagasta, Chile ii DEDICATORIA Esta tesis está dedicada al Señor, a mis amados mis padres y hermano. iii AGRADECIMIENTOS Al concluir esta etapa, deseo agradecer al Señor por su amor, ayuda y por darme la fortaleza de enfrentar este gran desafío y poder finalizarlo con éxito. A mis amados padres Ivón Velásquez Vielma y Daniel Ruiz Sandoval, y a mi hermano Daniel Ruiz Velásquez por estar siempre presentes, constante preocupación, y apoyo incondicionalmente durante todos estos años, con sus palabras de afecto y gran amor, gracias por todo. La realización de esta Tesis no hubiese sido posible sin el apoyo incondicional de mi profesor tutor (patrocinante), Dr. Manuel Zapata Arcos, por creer en mí y en este proyecto, por estar siempre a mi lado instándome a seguir adelante, brindándome su confianza, conocimiento, paciencia, consejo y amistad, gracias por todo profesor. Deseo agradecer a la Dra. María Teresa Gonzáles, por su confianza, conocimiento y por su constante apoyo para sacar esta iniciativa adelante, gracias profesora. A la profesora Mariella Rivas, por su gran apoyo, y por su gran amabilidad de ayudarme en las actividades metodológicas. También, deseo agradecer a Daniel Ordenes, Profesor Juan Morales, Maritza Fajardo, Espiridion Montanares, y al grupo de Laboratorio Bioal (Debra, Daniela, Javiera, Alejandro, Mari Carmen, Cristián, David, Alejandra, Lun) por la gran ayuda brindada para el desarrollo de esta tesis, amistad y por sus palabras de aliento. También, extiendo mis agradecimiento a Mauricio Escalona, quién estuvo siempre presente apoyándome, instándome a seguir adelante y quién se convirtió en un gran amigo de Tesis. Además, deseo extender mis agradecimientos hacia todas las personas que de una u otra forma estuvieron presentes durante todos estos años, en especial a Jessica, José, Nair, Yuri, Nancy, Sidgrid, Diony, Arianna y Mauro, y a mis familiares, amigos y compañeros. Deseo agradecer a la Compañía Santa María Spa., especialmente, al Sr. Pedro Granic, y a todo el personal, por su apoyo en las actividades de campo. iv Finalmente, deseo agradecer al apoyo brindado por Conicyt para el desarrollo de esta tesis, mediante la adjudicación de la Beca “Tesis de Doctorado en la Industria” y la Beca asignación anual para gastos operacionales del proyecto de Tesis Doctoral, también de Conicyt. v TABLA DE CONTENIDOS Página Calificaciones ………………………….………..……………….......................................ii Dedicatoria ……..……………………….………..………………...................................iii Agradecimientos ……………………….………..……………….....................................iv Tabla de contenidos …………………….………..………...……....................................vi Lista de tablas ………………………….………..………………....................................ix Lista de figuras …………………………..………..……………......................................ix Resumen ..….………...…………………………………………..……….........................xi Abstract .…………….………………………..…………...……….……........................xii Introducción ………………………………………………...............................................1 Características de la sexualidad en los moluscos bivalvos …...................................1 Determinación sexual en los moluscos bivalvos …………………………...............2 Diferenciación sexual o gonadal en moluscos bivalvos …………………................4 Esteroides sexuales en los moluscos bivalvos ………………………......................6 Tipos de esteroides sexuales ….………….………....................................................6 Función de los esteroides sexuales en la reproducción …..…………….…………...7 Función de los esteroides sexuales en la diferenciación sexual .…..…………….....8 Citocromo P450 aromatasa ....………………….....................................................10 Modelo biológico ………...….…………................................................................12 Objetivo de la tesis ……….…………………….....................................................16 Hipótesis ..…………….…….……………………...................................................18 Capítulo I. Sexual differentiation and size at first maturity of the mussel Choromytilus chorus (Molina, 1782) (Mollusca, Bivalvia) in Northern of Chile ……………………………………………...............................................................19 Abstract …………………………………………………..................................................20 Introduction and objectives ……..………………………..................................................21 Metodology ……………………..………………………..................................................24 Sampling ……….……..………………………......................................................24 Sex ratio ….…….……..…………………………..................................................25 Sexual differentiation ....…………………………...................................................25 Sexual maturity stages ....…………………………..................................................25 Succes of fertilization of the gametes of C. chorus …............................................27 Determination of the size at first sexual maturity …...............................................29 Statistical Analysis …………..…………………………...................................................29 Results …...……………………..………………………...................................................30 Sex ratio ……….………..………………………...................................................30 Sexual differentiation .…..………………………...................................................30 Histological analysis ..…..………………………...................................................31 Sexual maturity ..………….………………………................................................32 vi Size at first sexual maturity ………………………………….................................32 Success of fertilization of the gametes of C. chorus …….……..............................33 Discussion ……….……………..………………………...................................................35 Conclusion ……….……………..………………………...................................................40 Acknoledgements ..……………..………………………...................................................41 Literature cited …..……………..………………………...................................................42 Annex chapter I ………………………………………………….……….........................47 Capítulo II. Role of sex steroids in gonadal differentiation of the mussel Choromytilus chorus (Bivalvia Mytilidae) (Molina 1782) (parte I) ……………………..57 Abstract ………………………………………………………………..............................58 Introduction ………………..…………………………………………..............................59 Material and methods .……..…………………………………………..............................63 Collection of the animals and maintenance …....……….……………………......79 Maintenance of animals to experimental cages ……………………....................63 Exposure experiment to sex steroids …..…..…………………….........................64 Sampling ……………...……………....…..……….…………….........................65 Histology ……………...……………....…..……….…………….........................66 Sexual maturity stages and gonad development …………………………………66 Experiment to determine the success of fertilization of the gametes ....................67 Statistical analysis …....…………………………………………..........................68 Results ………………………………………………...…….............................................69 Treatments effects on survival and biometrics measures ….…..............................69 Effect of sex steroids on differentiation …….........................................................69 Effect of sex steroids on sexual maturity ……………...........................................70 Effect of steroids on the histology of the gonad ……...…………………….……72 Experiment to determine the viability of gametes .................................................73 Discussion ……..…………..…………………………………………..............................74 Effect on gonadal development in mussels ….......................................................74 Effect of sex steroids on sexual differentiation ……….……................................75 . Success of fertilization of the gametes …………………….…….........................78 Conclusion ………………………………………………………………………..............80 Acknowledgments ..............................................................................................................80 Literature cited …………...................................................................................................81 Annex chapter II ……..……................................................................................................86
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