Análisis transcriptómico para identificar genes biomarcadores que respondan ante la exposición al insecticida clorpirifos, en el pez dulceacuícola centroamericano Poecilia gillii
Fecha
2025
Autores
Sánchez González, Daniel
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Editor
Universidad Nacional (Costa Rica)
Resumen
Resumen. El uso excesivo de clorpirifos (CPF) en países tropicales como Costa Rica representa un posible riesgo para los ecosistemas de agua dulce. Este estudio investigó los efectos del CPF a una concentración ambientalmente relevante sobre la especie de pez nativa Poecilia gillii, empleando un enfoque integral que evaluó múltiples niveles de organización biológica. Utilizando RT-qPCR, se cuantificaron los cambios transcriptómicos en genes involucrados en diversos procesos biológicos, incluidos la inflamación y la apoptosis; annexin A1 (anxa1b), regulación de citoquinas; cytokine-inducible SH2-containing protein (cish), reacciones redox; NADH oxidoreductase subunit A2 (ndufa2), translocación de proteínas; Sec61 gamma subunit (sec61g), y biotransformación; glutathione S-transferase rho (gstr). Además, se evaluaron biomarcadores bioquímicos como enzimas de biotransformación de fase I; 7-ethoxyresorufin-O-deethylase (EROD) y fase II; glutathione S-transferase (GST), marcadores de estrés oxidativo; catalase (CAT) y lipid peroxidation (LPO) y la neurotoxicidad midiendo la actividad de cholinesterase (ChE) en tejidos cerebrales y musculares. Además, se realizaron pruebas de comportamiento evaluando la natación y las reacciones de antipredación. Tras una exposición transitoria de 48 horas a 5.5 µg/L de CPF, se observó una regulación significativa a la baja de los genes sec61g y gstr, una disminución en la actividad de CAT y efectos neurotóxicos evidenciados por la reducción de la actividad de ChE en los músculos. Aunque no se detectaron cambios conductuales significativos, nuestros resultados sugieren que la exposición a corto plazo a concentraciones ambientales relevantes de CPF puede alterar la expresión génica, comprometiendo la biotransformación y la síntesis de proteínas en los juveniles de P. gillii. Además, la neurotoxicidad observada, consistente con el mecanismo de acción del CPF, podría conducir a cambios conductuales sutiles. Este estudio proporciona evidencia de los efectos subletales del CPF en organismos no objetivos, destacando la importancia de considerar los cambios en la expresión génica en la evaluación de la toxicidad del CPF.
Summary. The excessive use of chlorpyrifos (CPF) in tropical countries such as Costa Rica represents a potential risk to freshwater ecosystems. This study investigated the effects of CPF at an environmentally relevant concentration on the native fish species Poecilia gillii, using an integrated approach that evaluated multiple levels of biological organization. Using RT-qPCR, transcriptomic changes were quantified in genes involved in a variety of biological processes, including in various biological processes, including inflammation and apoptosis; annexin A1 (anxa1b), cytokine regulation; cytokine-inducible SH2-containing protein (cish), redox reactions; NADH oxidoreductase subunit A2 (ndufa2), protein translocation; Sec61 gamma subunit (sec61g), and biotransformation; glutathione S-transferase rho (gstr). In addition, biochemical biomarkers were evaluated such as biotransformation enzymes of phase I; 7-ethoxyresorufin-O-deethylase (EROD) and phase II; glutathione S-transferase (GST), oxidative stress markers; catalase (CAT) and lipid peroxidation (LPO) and neurotoxicity by measuring cholinesterase (ChE) activity in brain and muscle tissues. In addition, behavioral tests were performed by evaluating swimming and anti-predation reactions. After a 48-hour transient exposure to 5.5 µg/L CPF, significant down-regulation of the sec61g and gstr genes, a decrease in CAT activity and neurotoxic effects CAT activity and neurotoxic effects evidenced by reduced ChE activity in muscles. Activity in muscles. Although no significant behavioral changes were detected, our results suggest that short-term exposure to relevant environmental concentrations of CPF may alter the CPF can alter gene expression, compromising biotransformation and protein synthesis in and protein synthesis in juvenile P. gillii. In addition, the observed neurotoxicity, consistent with the mechanism of action of CPF, could lead to subtle behavioral changes. changes. This study provides evidence for the sublethal effects of CPF in non-target organisms, highlighting the importance of non-target organisms, highlighting the importance of considering changes in gene expression in the assessment of CPF toxicity.
Summary. The excessive use of chlorpyrifos (CPF) in tropical countries such as Costa Rica represents a potential risk to freshwater ecosystems. This study investigated the effects of CPF at an environmentally relevant concentration on the native fish species Poecilia gillii, using an integrated approach that evaluated multiple levels of biological organization. Using RT-qPCR, transcriptomic changes were quantified in genes involved in a variety of biological processes, including in various biological processes, including inflammation and apoptosis; annexin A1 (anxa1b), cytokine regulation; cytokine-inducible SH2-containing protein (cish), redox reactions; NADH oxidoreductase subunit A2 (ndufa2), protein translocation; Sec61 gamma subunit (sec61g), and biotransformation; glutathione S-transferase rho (gstr). In addition, biochemical biomarkers were evaluated such as biotransformation enzymes of phase I; 7-ethoxyresorufin-O-deethylase (EROD) and phase II; glutathione S-transferase (GST), oxidative stress markers; catalase (CAT) and lipid peroxidation (LPO) and neurotoxicity by measuring cholinesterase (ChE) activity in brain and muscle tissues. In addition, behavioral tests were performed by evaluating swimming and anti-predation reactions. After a 48-hour transient exposure to 5.5 µg/L CPF, significant down-regulation of the sec61g and gstr genes, a decrease in CAT activity and neurotoxic effects CAT activity and neurotoxic effects evidenced by reduced ChE activity in muscles. Activity in muscles. Although no significant behavioral changes were detected, our results suggest that short-term exposure to relevant environmental concentrations of CPF may alter the CPF can alter gene expression, compromising biotransformation and protein synthesis in and protein synthesis in juvenile P. gillii. In addition, the observed neurotoxicity, consistent with the mechanism of action of CPF, could lead to subtle behavioral changes. changes. This study provides evidence for the sublethal effects of CPF in non-target organisms, highlighting the importance of non-target organisms, highlighting the importance of considering changes in gene expression in the assessment of CPF toxicity.
Descripción
Sánchez González, D. (2025). Análisis transcriptómico para identificar genes biomarcadores que respondan ante la exposición al insecticida clorpirifos, en el pez dulceacuícola centroamericano Poecilia gillii. [Tesis de Licenciatura]. Universidad Nacional, Heredia, Costa Rica
Palabras clave
PECES, GENES, ESTRÉS, ECOSISTEMAS, TOXICOLOGÍA, SUSTANCIAS TÓXICAS, TOXICOLOGY, AGUA DULCE, SWEET WATER, COSTA RICA