title: Proteomic changes occurring in the malaria mosquitoes Anopheles gambiae and Anopheles stephensi during aging.
abstract: The age of mosquitoes is a crucial determinant of their ability to transmit pathogens and their resistance to insecticides. We investigated changes to the abundance of proteins found in heads and thoraces of the malaria mosquitoes Anopheles gambiae and Anopheles stephensi as they aged. Protein expression changes were assessed using two-dimensional difference gel electrophoresis and the identity of differentially expressed proteins was determined by using either matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry or capillary high-pressure liquid chromatography coupled with a linear ion-trap (LTQ)-Orbitrap XL hybrid mass spectrometer. Protein biomarkers were validated by semi quantitative Western blot analysis. Nineteen and nine age dependent protein spots were identified for A. stephensi and A. gambiae, respectively. Among the proteins down-regulated with age were homologs of ADF/Cofilin, cytochome c1, heat shock protein-70 and eukaryotic translation initiation factor 5A (eIF5a). Proteins up-regulated with age included probable methylmalonate-semialdehyde dehydrogenase, voltage-dependent anion-selective channel and fructose bisphosphate aldolase. Semi quantitative Western blot analysis confirmed expression patterns observed by 2-D DIGE for eIF5a and ADF/Cofilin. Further work is recommended to determine whether these biomarkers are robust to infection, blood feeding and insecticide resistance. Robust biomarkers could then be incorporated into rapid diagnostic assays for ecological and epidemiological studies.
  
  
  CONCLUSIONS
  In this study, we have identified several proteins with characteristic changes in abundance in both A. gambiae and A. stephensi during their aging process. These changes may highlight underlying mechanisms beneath the relationship between mosquito age and factors affecting Plasmodium transmission and mosquito control. The similarity of changes in protein abundance between these species and the primary dengue vector Aedes aegypti, has revealed conserved patterns of aging-specific protein regulation.
authors.affiliation: Mosquito Control Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia. Electronic address: Maggy.Sikulu@qimrberghofer.edu.au.
authors.forename: Maggy T
authors.initials: MT
authors.lastname: Sikulu
authors.affiliation: Mosquito Control Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia. Electronic address: James.monkman@gmail.com.
authors.forename: James
authors.initials: J
authors.lastname: Monkman
authors.affiliation: The Protein Discovery Centre, QIMR Berghofer Medical Research Institute, Brisbane, Australia. Electronic address: Keyur.Dave@qimrberghofer.edu.au.
authors.forename: Keyur A
authors.initials: KA
authors.lastname: Dave
authors.affiliation: The Protein Discovery Centre, QIMR Berghofer Medical Research Institute, Brisbane, Australia. Electronic address: Marcus.Hastie@qimrberghofer.edu.au.
authors.forename: Marcus L
authors.initials: ML
authors.lastname: Hastie
authors.affiliation: Environmental Research Institute and Griffith School of Environment, Griffith University, Brisbane, Queensland, Australia. Electronic address: P.dale@griffith.edu.au.
authors.forename: Patricia E
authors.initials: PE
authors.lastname: Dale
authors.affiliation: Environmental Research Institute and Griffith School of Environment, Griffith University, Brisbane, Queensland, Australia. Electronic address: R.kitching@griffith.edu.au.
authors.forename: Roger L
authors.initials: RL
authors.lastname: Kitching
authors.affiliation: Environmental Health and Ecological Sciences Thematic Group, Ifakara Health Institute, Ifakara, United Republic of Tanzania; Vector Biology Department, Liverpool School of Tropical Medicine, Liverpool, United Kingdom. Electronic address: gkilleen@ihi.or.tz.
authors.forename: Gerry F
authors.initials: GF
authors.lastname: Killeen
authors.affiliation: Mosquito Control Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia. Electronic address: Brian.Kay@qimrberghofer.edu.au.
authors.forename: Brian H
authors.initials: BH
authors.lastname: Kay
authors.affiliation: The Protein Discovery Centre, QIMR Berghofer Medical Research Institute, Brisbane, Australia. Electronic address: Jeffery.Gorman@qimrberghofer.edu.au.
authors.forename: Jeffery J
authors.initials: JJ
authors.lastname: Gorman
authors.affiliation: Mosquito Control Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia. Electronic address: Leon.Hugo@qimrberghofer.edu.au.
authors.forename: Leon E
authors.initials: LE
authors.lastname: Hugo
mesh_terms: D000375:Aging / Q000502:physiology*; D000818:Animals; D000852:Anopheles / Q000378:metabolism* / Q000469:parasitology; D005786:Gene Expression Regulation / Q000502:physiology*; D019476:Insect Proteins / Q000096:biosynthesis*; D008288:Malaria / Q000635:transmission; D010961:Plasmodium; D040901:Proteomics
chemical_list: D019476:Insect Proteins
keywords: 2D-DIGE; A. stephensi; Age associated protein biomarkers; Anopheles gambiae s.s; Mass spectrometry; Semi quantitative Western blot analysis