Location: Mosquito and Fly Research Unit
Title: Cytochrome c gene and protein expression: developmental regulation, environmental response and pesticide sensitivity in the Aedes aegypti Authors
Submitted to: Journal of Medical Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 24, 2008
Publication Date: May 1, 2008
Citation: Zhao, L., Pridgeon, J.W., Becnel, J.J., Clark, G.G., Linthicum, K. 2008. Cytochrome c gene and protein expression: developmental regulation, environmental response and pesticide sensitivity in the Aedes aegypti. Journal of Medical Entomology. 45(3):401-408. Interpretive Summary: Cytochrome C is a highly conserved protein, found in multicellular organisms including animals, and also found in many unicellular organisms. Cytochrome C is a critical intermediate in apoptosis which is a controlled form of cell death used to kill cells in the process of development. Cytochrome c (AaeCytC) was recently studied from Aedes aegypti. To detect whether AaeCytC is developmentally regulated, we used real-time PCR to examine AaeCytC gene expression levels in different developmental stages of Aedes argypti. Real-time PCR showed that AaeCytC was highly expressed in teneral adult of Aedes aegypti. We also investigated that effect of environmental factors on AaeCytC gene expression, such as temperature, UV-light, and pesticide. Meanwhile, we also used Western blot analyses to examine the AeaCYTC protein levels in the developmental stages and effect of environmental factors. In conclusion, the expressions of AeaCytC gene and AeaCYTC protein in the life cycle of Ae. aegypti were highly regulated developmentally and environmentally. Cytochrome c activity has, for the first time, been examined in detail for all developmental stages of Ae. aegypti. The present study suggests that AeaCytC plays an important functional role in the development of Ae. aegypti and may provide information useful for designing novel control strategies for mosquitoes and identifying new pathways to target for the development of molecular pesticides.
Technical Abstract: Cytochrome c is a highly conserved protein that is found in many multicellular and unicellular organisms. Cytochrome c is a critical intermediate in apoptosis: a controlled form of cell death that kills cells as part of their natural process of development and in response to environmental condition. To detect whether cytochrome c of the mosquito Aedes aegypti (AeaCytC) is developmentally regulated, we employed quantitative real-time PCR to examine AeaCytC gene expression levels in different developmental stages of Ae. aegypti. Quantitative real-time PCR showed that AeaCytC was expressed in each developmental stage, at different points in time, and was highly expressed in teneral female Ae. aegypti. Ae. aegypti cytochrome c protein (AeaCYTC) was detected only in adult mosquitoes, not in early developmental stages of Ae. aegypti. We also investigated the effect of certain environmental factors (e.g. temperature, UV-light and permethrin insecticide) on AeaCytC gene and AeaCYTC protein expression showing different responses in ages. Taken together, these results suggest that AeaCytC gene and AeaCYTC protein both play functional roles in the development of Ae. aegypti and its response to environmental stress and pesticide challenge.