Evaluación del efecto del estrés por manejo en la expresión de genes asociados a la inmunidad del congrio colorado (Genypterus Chilensis) mediante RNA-seq
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2017
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es
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Universidad Andrés Bello
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Licencia CC
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El congrio colorado (Genypterus chilensis) es una especie endémica propuesta para la diversificación del mercado acuícola chileno. El Centro de Investigación Marina de Quintay (CIMARQ) muestra los primeros avances en relación al cultivo intensivo de este ejemplar. Desafortunadamente, uno de los principales problemas asociados al cultivo intensivo de peces es el estrés generado durante el desarrollo de esta actividad, siendo el estrés por manejo uno de los más comunes. Se ha reportado que el estrés afecta de diferentes formas la inmunidad de los teleósteos, lo que pone en riesgo la supervivencia de estos animales durante su cultivo.
Hoy en día aún existen diversas interrogantes respecto a cómo el estrés puede afectar la inmunidad de los peces, y más aún en congrio colorado. Además, la información biológica relacionada con este teleósteo es escasa y pesar de que el primer transcriptoma de referencia secuenciado a partir de muestras de tejido muscular y hepático fue recientemente anotado, éste no entregó datos relevantes respecto a la expresión de genes asociados a respuesta inmune durante la respuesta a estrés. De esta manera, el objetivo de este trabajo es analizar la expresión diferencial de genes asociados a la regulación del sistema inmune del congrio colorado en respuesta al estrés por manejo. Para esto se ensambló de novo el transcriptoma de riñón anterior de congrio colorado, principal órgano inmunitario en peces. Este está conformado por 86.840 secuencias, de las cuales el 42% tiene anotación funcional. El estudio de expresión in silico (RNA-seq) permitió identificar 1.094 genes expresados diferencialmente. Se validó la expresión de algunos de éstos genes mediante RT-qPCR. El análisis de enriquecimiento ontológico reveló que procesos biológicos como la respuesta a glucocorticoides, sistema circulatorio y sistema inmune se veían enriquecidos en la condición de estrés, mientras que procesos relacionados con el metabolismo, especialmente el catabolismo y la biosíntesis de distintas moléculas como nucleótidos y aminoácidos, entre otras, se encontraron reprimidos.
Los hallazgos recopilados en el presente trabajo enriquecen los conocimientos relacionados con el efecto que genera el estrés por manejo, una condición inherente de los cultivos, en la fisiología de este organismo, particularmente en su respuesta inmune. Ésta nueva información se podría utilizar en futuros planes de manejo para finalmente concretar un plan de cultivo de congrio colorado con proyecciones a largo plazo.
The red cusk-eel (Genypterus chilensis) is an endemic species with proposed potential in diversifying the Chilean aquaculture industry. While the Quintay Marine Research Center (CIMARQ) has made initial progress in the intensive cultivation of this species. Unfortunately, one of the main problems associated with intensive fish farming, is the stress generated during the development of this activity, being the handling stress one of the most common. It is particularly well-known that stress can differentially affect teleost immunity, thus endangering animal survival during cultivation. Nevertheless, a complete understanding of how stress affects fish immune systems remains lacking, and this knowledge gap is even more marked for the red cusk-eel, a species with overall limited biological information. Although the first reference transcriptome sequenced from G. chilensis muscle and liver tissue was recently annotated, this did not provide relevant data regarding gene expressions associated with a stress-induced immune response. Therefore, the aim of this study was to analyze the differential expression of genes related to immune regulation in response to handling stress. For this, the transcriptome of the G. chilensis anterior kidney, the main immunological organ in fish, was de novo assembled. The transcriptome contained 86,840 sequences, of which 42% were functionally annotated. In silico expression analysis through RNA-seq identified 1,094 differentially expressed genes, some of which were validated by RT-qPCR. Ontological enrichment analysis revealed that biological processes were enriched by stress, including the glucocorticoid, circulatory system, and immune system responses. In turn, processes related to metabolism were repressed, especially the catabolism and biosynthesis of different molecules, such as nucleotides and amino acids, among others. In conclusion, the obtained data provide novel knowledge related to the effects of handling stress, an inherent culture condition, on the physiology and immune response of the red cusk-eel. This information could be useful for future management and long-term planning for the intensive cultivation of G. chilensis.
The red cusk-eel (Genypterus chilensis) is an endemic species with proposed potential in diversifying the Chilean aquaculture industry. While the Quintay Marine Research Center (CIMARQ) has made initial progress in the intensive cultivation of this species. Unfortunately, one of the main problems associated with intensive fish farming, is the stress generated during the development of this activity, being the handling stress one of the most common. It is particularly well-known that stress can differentially affect teleost immunity, thus endangering animal survival during cultivation. Nevertheless, a complete understanding of how stress affects fish immune systems remains lacking, and this knowledge gap is even more marked for the red cusk-eel, a species with overall limited biological information. Although the first reference transcriptome sequenced from G. chilensis muscle and liver tissue was recently annotated, this did not provide relevant data regarding gene expressions associated with a stress-induced immune response. Therefore, the aim of this study was to analyze the differential expression of genes related to immune regulation in response to handling stress. For this, the transcriptome of the G. chilensis anterior kidney, the main immunological organ in fish, was de novo assembled. The transcriptome contained 86,840 sequences, of which 42% were functionally annotated. In silico expression analysis through RNA-seq identified 1,094 differentially expressed genes, some of which were validated by RT-qPCR. Ontological enrichment analysis revealed that biological processes were enriched by stress, including the glucocorticoid, circulatory system, and immune system responses. In turn, processes related to metabolism were repressed, especially the catabolism and biosynthesis of different molecules, such as nucleotides and amino acids, among others. In conclusion, the obtained data provide novel knowledge related to the effects of handling stress, an inherent culture condition, on the physiology and immune response of the red cusk-eel. This information could be useful for future management and long-term planning for the intensive cultivation of G. chilensis.
Notas
Tesis (Magíster en Biotecnología)
Palabras clave
Estrés en Peces, Congrio, Enfermedades