Evaluación de los componentes de la vía ILPs (insulin like-peptides) durante restricción nutricional en el Mejillón chileno (Mytilus chilensis)
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2017
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es
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Universidad Andrés Bello
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Licencia CC
Licencia CC
Resumen
Chile se posiciona entre los 5 principales productores de mejillones del mundo, concentrando un gran porcentaje de la producción en el mejillón chileno (Mytilus chilensis). Para el desarrollo de la miticultura la tasa de crecimiento de ejemplares juega un rol fundamental, sin embargo, diversos factores pueden estar afectando su crecimiento, como por ejemplo la disponibilidad de alimentos. El crecimiento en vertebrados como invertebrados está regulado principalmente por el sistema de factor de crecimiento tipo insulina (IGF), el cual es una vía de señalización muy conservada. A pesar de la gran cantidad de publicaciones relacionadas con este sistema en vertebrados y su modulación bajo diferentes estados nutricionales, existen escasos reportes relacionados con la restricción de alimentos y su impacto en el mecanismo molecular relacionado con las vías de señalización gatillada por péptidos tipo insulina (ILPs) en mejillones. Por lo tanto, el objetivo de este trabajo fue evaluar los efectos de la restricción nutricional en la expresión de componentes claves de la vía ILPs en M. chilensis. Los mejillones fueron sometidos a un protocolo de restricción nutricional, además de un tratamiento utilizando un inhibidor de la vía de IGF-1 (picropodofilina). Se realizó la extracción RNA total del manto en M. chilensis para realizar una transcripción reversa cuantitativa de: péptidos tipo insulina (ilp); fosfoinositol 3-quinasa (pi3k); proteína quinasa b (akt), forkhead box G1 (foxg1) y e3 ubiquitina-proteína ligasas (fbxo25 y trim56), además la extracción proteica y western blot de la forma fosforilada del receptor de IGF-1 y el contenido de proteínas ubiquitinadas en las diferentes condiciones a las 3 semanas. El efecto de la restricción nutricional, al igual que el tratamiento con picropodofilina induce una pérdida de peso en los mejillones, lo que se correlaciona con la regulación negativa del sistema ILPs. Nuestros resultados sugieren que el crecimiento está estrechamente relacionado con la vía de señalización desencadenada por ILPs, modulando procesos anabólicos y catabólicos en M. chilensis.
Chile is among the top five mussel producers in the world, concentrating a large percentage of the production of the “Mejillón Chileno” (Mytilus chilensis). For the development of the miticulture the rate of growth of individuals plays a fundamental role, however, several factors may be affecting its growth, such as food availability. Growth in vertebrates as invertebrates is mainly regulated by the insulin-like growth factor (IGF) system, which is a highly conserved signaling pathway. Despite the large number of publications related to this system in vertebrates and their modulation under different nutritional states, there are few reports related to food restriction and its impact on the molecular mechanism related to signaling pathways triggered by insulin-like peptides (ILPs) in mussels. Therefore, the objective of this work was to evaluate the effects of nutritional restriction on the expression of key components of the ILPs pathway in M. chilensis. Mussels were subjected to a nutritional restriction protocol, in addition to a treatment using an inhibitor of the IGF-1 pathway (picropodophyllin). Total mantle RNA extraction was performed in M. chilensis to develop a quantitative reverse transcription of: insulin-like peptides (ilp); Phosphoinositol 3-kinase (pi3k); Protein kinase b (akt), forkhead box G1 (foxg1) and e3 ubiquitin-protein ligases (fbxo25 and trim56), in addition protein and western blot extraction of the phosphorylated form of the IGF-1 receptor and the ubiquitinated protein content in the different conditions at 3 weeks. The effect of nutritional restriction, as well as the treatment with picropodophyllin induces weight loss in mussels, which correlates with the down-regulation of the ILPs system. Our results suggest that the growth is closely related to the signaling pathway triggered by ILPs, modulating anabolic and catabolic processes in M. chilensis.
Chile is among the top five mussel producers in the world, concentrating a large percentage of the production of the “Mejillón Chileno” (Mytilus chilensis). For the development of the miticulture the rate of growth of individuals plays a fundamental role, however, several factors may be affecting its growth, such as food availability. Growth in vertebrates as invertebrates is mainly regulated by the insulin-like growth factor (IGF) system, which is a highly conserved signaling pathway. Despite the large number of publications related to this system in vertebrates and their modulation under different nutritional states, there are few reports related to food restriction and its impact on the molecular mechanism related to signaling pathways triggered by insulin-like peptides (ILPs) in mussels. Therefore, the objective of this work was to evaluate the effects of nutritional restriction on the expression of key components of the ILPs pathway in M. chilensis. Mussels were subjected to a nutritional restriction protocol, in addition to a treatment using an inhibitor of the IGF-1 pathway (picropodophyllin). Total mantle RNA extraction was performed in M. chilensis to develop a quantitative reverse transcription of: insulin-like peptides (ilp); Phosphoinositol 3-kinase (pi3k); Protein kinase b (akt), forkhead box G1 (foxg1) and e3 ubiquitin-protein ligases (fbxo25 and trim56), in addition protein and western blot extraction of the phosphorylated form of the IGF-1 receptor and the ubiquitinated protein content in the different conditions at 3 weeks. The effect of nutritional restriction, as well as the treatment with picropodophyllin induces weight loss in mussels, which correlates with the down-regulation of the ILPs system. Our results suggest that the growth is closely related to the signaling pathway triggered by ILPs, modulating anabolic and catabolic processes in M. chilensis.
Notas
Tesis (Magíster en Biotecnología)
Palabras clave
Mejillones, Análisis