Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus

BACKGROUND: Temporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characteri...

Descripción completa

Detalles Bibliográficos
Autores principales: Puthiyakunnon, Santhosh, Yao, Yunying, Li, Yiji, Gu, Jinbao, Peng, Hongjuan, Chen, Xiaoguang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3750763/
https://www.ncbi.nlm.nih.gov/pubmed/23924583
http://dx.doi.org/10.1186/1756-3305-6-230
_version_ 1782281475937271808
author Puthiyakunnon, Santhosh
Yao, Yunying
Li, Yiji
Gu, Jinbao
Peng, Hongjuan
Chen, Xiaoguang
author_facet Puthiyakunnon, Santhosh
Yao, Yunying
Li, Yiji
Gu, Jinbao
Peng, Hongjuan
Chen, Xiaoguang
author_sort Puthiyakunnon, Santhosh
collection PubMed
description BACKGROUND: Temporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characterization of these miRNAs was not known. Accordingly our study evaluated the functional characterization of three miRNAs, which are temporally up-regulated in the various developmental stages of Ae. albopictus mosquitoes. METHODS: miRNA mimics, inhibitors and negative controls were designed and their knock-in and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting in their specific developmental stages. The functional role of each individual miRNA was analyzed with various parameters of development such as, hatching rate and hatching time in embryos, eclosion rate in larvae, longevity and fecundity in the adult mosquitoes. RESULTS: The knock-in with the specifically designed miRNA mimics showed increased levels of expression of miRNA compared with their normal controls. We confirmed these findings using qRT-PCR, both by in vitro expression in C6/36 mosquito cell lines after transfection as well as in in vivo expression in developmental stages of mosquitoes by microinjection. The knock-down of expression with the corresponding inhibitors showed a considerable decrease in the expression levels of these miRNAs and obvious functional effects in Ae. albopictus development, detected by a decrease in the hatching rate of embryos and eclosion rate in larvae and a marked reduction in longevity and fecundity in adults. CONCLUSION: This study carried out by knock-in and knock-down of specifically and temporally expressed miRNAs in Ae. albopictus by microinjection is a novel study to delineate the importance of the miRNA expression in regulating mosquito development. The knock-down and loss of function of endogenously expressed miRNAs by the miRNA inhibitors in specific developmental stages had considerable effects on development, but enhancement of their gain of function was not observed on knock-in of these specific miRNAs. Hence, our study indicates that an optimal level of endogenous expression of miRNA is indispensable for the normal development and maintenance of the vectorial population density and pathogen transmissibility of this mosquito vector.
format Online
Article
Text
id pubmed-3750763
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-37507632013-08-24 Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus Puthiyakunnon, Santhosh Yao, Yunying Li, Yiji Gu, Jinbao Peng, Hongjuan Chen, Xiaoguang Parasit Vectors Research BACKGROUND: Temporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characterization of these miRNAs was not known. Accordingly our study evaluated the functional characterization of three miRNAs, which are temporally up-regulated in the various developmental stages of Ae. albopictus mosquitoes. METHODS: miRNA mimics, inhibitors and negative controls were designed and their knock-in and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting in their specific developmental stages. The functional role of each individual miRNA was analyzed with various parameters of development such as, hatching rate and hatching time in embryos, eclosion rate in larvae, longevity and fecundity in the adult mosquitoes. RESULTS: The knock-in with the specifically designed miRNA mimics showed increased levels of expression of miRNA compared with their normal controls. We confirmed these findings using qRT-PCR, both by in vitro expression in C6/36 mosquito cell lines after transfection as well as in in vivo expression in developmental stages of mosquitoes by microinjection. The knock-down of expression with the corresponding inhibitors showed a considerable decrease in the expression levels of these miRNAs and obvious functional effects in Ae. albopictus development, detected by a decrease in the hatching rate of embryos and eclosion rate in larvae and a marked reduction in longevity and fecundity in adults. CONCLUSION: This study carried out by knock-in and knock-down of specifically and temporally expressed miRNAs in Ae. albopictus by microinjection is a novel study to delineate the importance of the miRNA expression in regulating mosquito development. The knock-down and loss of function of endogenously expressed miRNAs by the miRNA inhibitors in specific developmental stages had considerable effects on development, but enhancement of their gain of function was not observed on knock-in of these specific miRNAs. Hence, our study indicates that an optimal level of endogenous expression of miRNA is indispensable for the normal development and maintenance of the vectorial population density and pathogen transmissibility of this mosquito vector. BioMed Central 2013-08-08 /pmc/articles/PMC3750763/ /pubmed/23924583 http://dx.doi.org/10.1186/1756-3305-6-230 Text en Copyright © 2013 Puthiyakunnon et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Puthiyakunnon, Santhosh
Yao, Yunying
Li, Yiji
Gu, Jinbao
Peng, Hongjuan
Chen, Xiaoguang
Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title_full Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title_fullStr Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title_full_unstemmed Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title_short Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus
title_sort functional characterization of three micrornas of the asian tiger mosquito, aedes albopictus
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3750763/
https://www.ncbi.nlm.nih.gov/pubmed/23924583
http://dx.doi.org/10.1186/1756-3305-6-230
work_keys_str_mv AT puthiyakunnonsanthosh functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus
AT yaoyunying functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus
AT liyiji functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus
AT gujinbao functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus
AT penghongjuan functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus
AT chenxiaoguang functionalcharacterizationofthreemicrornasoftheasiantigermosquitoaedesalbopictus