Analysis of cycle gene expression in Aedes aegypti brains by in situ hybridization

• Published in: PloS one Vol. 8; no. 1; p. e52559
• Main Authors: Chahad-Ehlers, Samira, Gentile, Carla, Lima, José Bento Pereira, Peixoto, Alexandre Afranio, Bruno, Rafaela Vieira
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02-01-2013; Public Library of Science (PLoS)
• Subjects: Aedes - metabolism; Aedes aegypti; Animals; Anopheles gambiae; Aquatic insects; ARNTL Transcription Factors - genetics; ARNTL Transcription Factors - metabolism; Behavior; Biology; Brain; Brain - metabolism; Circadian Rhythm; Circadian rhythms; Culicidae; DNA, Complementary - metabolism; Drosophila; Drosophila melanogaster; Gene expression; Gene Expression Regulation; Genetic aspects; Genomics; Host searching behavior; Hybridization; In Situ Hybridization; Insect Proteins - genetics; Insect Proteins - metabolism; Insects; Laboratories; Lobes; Male; Medicine; Molecular modelling; Mosquitoes; Neurons - metabolism; Oligonucleotide Probes - metabolism; Physiological aspects; Polymerase Chain Reaction; Proteins; RNA, Messenger - metabolism; Staining; Time Factors; Tropical diseases; Vectors; Brazil; Rio de Janeiro Brazil
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0052559

Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors, relatively little is known about the molecular mechanisms underlying processes involved in the temporal pattern of its activity and host seeking behavior. In this study, we analyzed the expression of the cycle (cyc) gene, one of the core components of the circadian clock, in Ae. aegypti brains by in situ hybridization at two different time points in light-dark conditions and compared the results with those obtained using a quantitative PCR assay (qPCR). Within the brain, differential labeling was detected according to distinct areas empirically pre-defined. Six out of seven of these areas showed significantly higher staining at ZT3 (three hours after light-on) compared to ZT11 (one before light-off), which is consistent with the qPCR data. Predominant staining was observed in three of those areas which correspond to positions of the optical and antennal lobes, as well as the region where the neurons controlling activity rhythms are presumably localized.