ArticleParasites & vectors2015
Evaluation of reference genes for insect olfaction studies.
Article in Parasites & vectors, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 38 citations in OpenAlex.
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- Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis inInsects · 2024Article
- Identification and validation of reference genes for normalization of gene expression analysis using qRT-PCR inFrontiers in physiology · 2023Article
- Differentiation of action mechanisms between natural and synthetic repellents through neuronal electroantennogram and proteomic in Aedes aegypti (Diptera: Culicidae).Scientific reports · 2022Article
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- Characterization of ThreeFrontiers in plant science · 2022Article
- Identification of suitable reference genes for expression profiling studies using qRT-PCR in an important insect pest, Maruca vitrata.Molecular biology reports · 2021Article
- MiR-279-3p regulates deltamethrin resistance through CYP325BB1 in Culex pipiens pallens.Parasites & vectors · 2021Article
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- Selection of candidate reference genes for RT-qPCR analysis in Argulus siamensis and their validation through screening of drugs and drug targets.Scientific reports · 2019Article
- Identification of reference genes for RT-qPCR data normalization in Gammarus fossarum (Crustacea Amphipoda).Scientific reports · 2018Article
- Selection of Reference Genes for the Normalization of RT-qPCR Data in Gene Expression Studies in Insects: A Systematic Review.Frontiers in physiology · 2018Article
- Comparative and functional triatomine genomics reveals reductions and expansions in insecticide resistance-related gene families.PLoS neglected tropical diseases · 2017Article
- Quantitative real-time PCR analysis ofPeerJ · 2017Article
- Functional development of carbon dioxide detection in the maxillary palp of Anopheles gambiae.The Journal of experimental biology · 2015Article
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Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundQuantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of 'housekeeping genes', involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori.
methodsWe evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: α-tubulin; β-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age,developmental stage and feeding status were tested in both sexes.
resultsWe show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards.
conclusionsThis work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.
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