Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans

The Chagas disease vector Triatoma infestans shows daily rhythms controlled by the biological clock genes. The RNA interference (RNAi) is a specific silencing mechanism that disrupts the gene expression. In order to analyze the effect of silencing the clock gene perio...

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Detalles Bibliográficos
Autores principales: Córdoba, LE, García, BA, Serradel, MC, Stroppa, MM
Formato: Artículo revista
Lenguaje:Español
Publicado: Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2023
Materias:
Chagas disease
Triatoma infestans
Chronobiology
period gene
Enfermedad de Chagas
Cronobiología
Gen Period
Acceso en línea:https://revistas.unc.edu.ar/index.php/med/article/view/42668
Aporte de:
Revista de la Facultad de Ciencias Médicas de Córdoba de Universidad Nacional de Córdoba
id I10-R327-article-42668
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institution Universidad Nacional de Córdoba
institution_str I-10
repository_str R-327
container_title_str Revista de la Facultad de Ciencias Médicas de Córdoba
language Español
format Artículo revista
topic Chagas disease
Triatoma infestans
Chronobiology
period gene
Enfermedad de Chagas
Triatoma infestans
Cronobiología
Gen Period
spellingShingle Chagas disease
Triatoma infestans
Chronobiology
period gene
Enfermedad de Chagas
Triatoma infestans
Cronobiología
Gen Period
Córdoba, LE
García, BA
Serradel, MC
Stroppa, MM
Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
topic_facet Chagas disease
Triatoma infestans
Chronobiology
period gene
Enfermedad de Chagas
Triatoma infestans
Cronobiología
Gen Period
author Córdoba, LE
García, BA
Serradel, MC
Stroppa, MM
author_facet Córdoba, LE
García, BA
Serradel, MC
Stroppa, MM
author_sort Córdoba, LE
title Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
title_short Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
title_full Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
title_fullStr Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
title_full_unstemmed Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans
title_sort effect of silencing the clock gene period in the chagas disease vector triatoma infestans
description The Chagas disease vector Triatoma infestans shows daily rhythms controlled by the biological clock genes. The RNA interference (RNAi) is a specific silencing mechanism that disrupts the gene expression. In order to analyze the effect of silencing the clock gene period (per) in T. infestans, RNAi protocols with different feeding regimens were used and its daily expression profile was analyzed in the nervous tissue. RNAi of the gene per and RNAi of the control gene β-Lactamase (β-Lac) were injected into adult female and male individuals. The dissections were performed every 6 hr over a 24-hour period and the time of day was reported as Zeitgeber Time (ZT). Samples consisted of groups of 3 individuals and 3 biological replicates were analyzed for each ZT. The expression of per gene was determined by Retrotranscription-Real Time PCR (RT-qPCR). The results were analyzed by One-way ANOVA test followed by the Bonferroni test (multiple comparison). Silencing of per gene in T. infestans males and females showed a significant decrease of expression at transcriptional level in the group injected with the RNAi of per  gene compared to the levels found in the control groups non-injected and injected with RNAi for β-Lac gene . The feeding regimens used in the protocols did not significantly affect the level of interference. On the other hand, the silencing of per gene reduced its expression in a half in all the ZTs analyzed and was not observed the characteristic daily variation at the transcriptional level that this clock gene presents. The results obtained allowed to establish that the silencing of per gene was effective in both males and females and was not affected by the feeding regimens used in the interference protocols. 
publisher Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
publishDate 2023
url https://revistas.unc.edu.ar/index.php/med/article/view/42668
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first_indexed 2024-09-03T21:04:45Z
last_indexed 2024-09-03T21:04:45Z
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spelling I10-R327-article-426682023-10-19T21:20:06Z Effect of silencing the clock gene period in the Chagas disease vector Triatoma infestans Efecto del silenciamiento del gen reloj period en el vector de la enfermedad de Chagas Triatoma infestans Córdoba, LE García, BA Serradel, MC Stroppa, MM Chagas disease Triatoma infestans Chronobiology period gene Enfermedad de Chagas Triatoma infestans Cronobiología Gen Period The Chagas disease vector Triatoma infestans shows daily rhythms controlled by the biological clock genes. The RNA interference (RNAi) is a specific silencing mechanism that disrupts the gene expression. In order to analyze the effect of silencing the clock gene period (per) in T. infestans, RNAi protocols with different feeding regimens were used and its daily expression profile was analyzed in the nervous tissue. RNAi of the gene per and RNAi of the control gene β-Lactamase (β-Lac) were injected into adult female and male individuals. The dissections were performed every 6 hr over a 24-hour period and the time of day was reported as Zeitgeber Time (ZT). Samples consisted of groups of 3 individuals and 3 biological replicates were analyzed for each ZT. The expression of per gene was determined by Retrotranscription-Real Time PCR (RT-qPCR). The results were analyzed by One-way ANOVA test followed by the Bonferroni test (multiple comparison). Silencing of per gene in T. infestans males and females showed a significant decrease of expression at transcriptional level in the group injected with the RNAi of per  gene compared to the levels found in the control groups non-injected and injected with RNAi for β-Lac gene . The feeding regimens used in the protocols did not significantly affect the level of interference. On the other hand, the silencing of per gene reduced its expression in a half in all the ZTs analyzed and was not observed the characteristic daily variation at the transcriptional level that this clock gene presents. The results obtained allowed to establish that the silencing of per gene was effective in both males and females and was not affected by the feeding regimens used in the interference protocols.  El vector de la enfermedad de Chagas Triatoma infestans muestra ritmos diarios controlados por los genes del reloj biológico. La interferencia por ARN (ARNi) es un mecanismo de silenciamiento génico que interrumpe la expresión de genes específicos. Para el silenciamiento del gen reloj period (per) se utilizaron protocolos de ARNi con diferentes esquemas de alimentación y se analizó el efecto del silenciamiento en su perfil de expresión diario en tejido nervioso de T. infestans. Los ARNi del gen per y del gen control β-Lactamasa (β-Lac) se inyectaron en ejemplares adultos y se realizaron disecciones cada 6 hs durante un período de 24 hs. La hora del día fue expresada en Zeitgeber Time (ZT).  Se utilizaron pooles de 3 individuos y 3 replicas experimentales por cada ZT.  En cada grupo se determinó la expresión del gen per mediante la técnica de Retrotranscripción y PCR en Tiempo Real (RT-qPCR).  Los resultados se analizaron mediante tests de ANOVA (One-way ANOVA) seguidos del Test de Bonferroni (de comparación múltiple). El silenciamiento del gen per en machos y hembras de T. infestans mostró una disminución significativa en la expresión transcripcional en el grupo de insectos inyectados con el ARNi del gen per con respecto a los niveles hallados en los grupos control (no inyectados e inyectados con el ARNi para el gen β-Lac). Los esquemas de alimentación utilizados en los protocolos no afectaron de forma significativa el nivel de interferencia. Por otra parte, el silenciamiento del gen per redujo su expresión en todos los ZT analizados y no se observaron las variaciones diarias características en la expresión a nivel transcripcional que presenta este gen reloj. Los resultados obtenidos permitieron establecer que el silenciamiento del gen per fue efectivo tanto en machos como en hembras y no se ve afectado por los esquemas de alimentación utilizados en los protocolos de interferencia. Por otra parte, el silenciamiento del gen per afectó el perfil de expresión del gen reduciendo a la mitad su expresión en todos los ZT analizados. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2023-10-19 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/42668 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 80 (2023): Suplemento JIC XXIV Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 80 (2023): Suplemento JIC XXIV Revista da Faculdade de Ciências Médicas de Córdoba; v. 80 (2023): Suplemento JIC XXIV 1853-0605 0014-6722 spa https://revistas.unc.edu.ar/index.php/med/article/view/42668/42898 Derechos de autor 2023 Universidad Nacional de Córdoba http://creativecommons.org/licenses/by-nc/4.0

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