title: Proteomic biomarkers for ageing the mosquito Aedes aegypti to determine risk of pathogen transmission.
abstract: Biomarkers of the age of mosquitoes are required to determine the risk of transmission of various pathogens as each pathogen undergoes a period of extrinsic incubation in the mosquito host. Using the 2-D Difference Gel Electrophoresis (2-D DIGE) procedure, we investigated the abundance of up to 898 proteins from the Yellow Fever and dengue virus vector, Aedes aegypti, during ageing. By applying a mixed-effects model of protein expression, we identified five common patterns of abundance change during ageing and demonstrated an age-related decrease in variance for four of these. This supported a search for specific proteins with abundance changes that remain tightly associated with ageing for use as ageing biomarkers. Using MALDI-TOF/TOF mass spectrometry we identified ten candidate proteins that satisfied strict biomarker discovery criteria (identified in two out of three multivariate analysis procedures and in two cohorts of mosquitoes). We validated the abundances of the four most suitable candidates (Actin depolymerising factor; ADF, Eukaryotic initiation factor 5A; eIF5A, insect cuticle protein Q17LN8, and Anterior fat body protein; AFP) using semi-quantitative Western analysis of individual mosquitoes of six ages. The redox-response protein Manganese superoxide dismutase (SOD2) and electron shuttling protein Electron transfer oxidoreductase (ETO) were subject to post-translational modifications affecting their charge states with potential effects on function. For the four candidates we show remarkably consistent decreases in abundance during ageing, validating initial selections. In particular, the abundance of AFP is an ideal biomarker candidate for whether a female mosquito has lived long enough to be capable of dengue virus transmission. We have demonstrated proteins to be a suitable class of ageing biomarkers in mosquitoes and have identified candidates for epidemiological studies of dengue and the evaluation of new disease reduction projects targeting mosquito longevity.
authors.affiliation: Mosquito Control Laboratory, Queensland Institute of Medical Research, Brisbane, Queensland, Australia. Leon.Hugo@qimr.edu.au
authors.forename: Leon E
authors.initials: LE
authors.lastname: Hugo
authors.forename: James
authors.initials: J
authors.lastname: Monkman
authors.forename: Keyur A
authors.initials: KA
authors.lastname: Dave
authors.forename: Leesa F
authors.initials: LF
authors.lastname: Wockner
authors.forename: Geoff W
authors.initials: GW
authors.lastname: Birrell
authors.forename: Emma L
authors.initials: EL
authors.lastname: Norris
authors.forename: Vivian J
authors.initials: VJ
authors.lastname: Kienzle
authors.forename: Maggy T
authors.initials: MT
authors.lastname: Sikulu
authors.forename: Peter A
authors.initials: PA
authors.lastname: Ryan
authors.forename: Jeffery J
authors.initials: JJ
authors.lastname: Gorman
authors.forename: Brian H
authors.initials: BH
authors.lastname: Kay
mesh_terms: D000330:Aedes / Q000378:metabolism*; D000375:Aging; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D005260:Female; D007303:Insect Vectors / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics
chemical_list: D015415:Biomarkers; D020543:Proteome
references.citation: PLoS One. 2011 Feb 09;6(2):e16545
references.pmid: 21347421
references.citation: Am J Trop Med Hyg. 1987 Jan;36(1):143-52
references.pmid: 3812879
references.citation: Bioinformatics. 2006 Jul 15;22(14):1745-52
references.pmid: 16675467
references.citation: Biochim Biophys Acta. 2010 Feb;1804(2):318-25
references.pmid: 19837190
references.citation: Lancet. 2012 Feb 18;379(9816):662-71
references.pmid: 22100854
references.citation: Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14355-60
references.pmid: 8962055
references.citation: Mol Cell Proteomics. 2012 May;11(5):108-27
references.pmid: 22322095
references.citation: Cell Cycle. 2011 Nov 1;10(21):3617-8
references.pmid: 22037286
references.citation: PLoS Biol. 2011 Sep;9(9):e1001144
references.pmid: 21909241
references.citation: Lancet. 2007 Nov 10;370(9599):1644-52
references.pmid: 17993365
references.citation: Electrophoresis. 2000 Mar;21(5):841-5
references.pmid: 10768767
references.citation: J Biol Chem. 1975 May 25;250(10):4007-21
references.pmid: 236308
references.citation: J Med Entomol. 1999 Nov;36(6):824-30
references.pmid: 10593086
references.citation: Adv Genet. 2003;49:67-109
references.pmid: 12779251
references.citation: Bioinformatics. 2005 May 1;21(9):2067-75
references.pmid: 15657102
references.citation: Biochemistry. 1985 Jul 16;24(15):3922-5
references.pmid: 2996585
references.citation: J Med Entomol. 2008 May;45(3):353-69
references.pmid: 18533427
references.citation: J Proteomics. 2011 Jan 1;74(1):1-18
references.pmid: 20656082
references.citation: Electrophoresis. 1997 Oct;18(11):2071-7
references.pmid: 9420172
references.citation: Science. 2005 Jun 10;308(5728):1641-2
references.pmid: 15947190
references.citation: Development. 2006 May;133(9):1789-97
references.pmid: 16571634
references.citation: Science. 2009 Jan 2;323(5910):141-4
references.pmid: 19119237
references.citation: Malar J. 2012 Jan 23;11:24
references.pmid: 22269002
references.citation: Annu Rev Entomol. 2010;55:207-25
references.pmid: 19725772
references.citation: J Cell Biol. 2009 Jun 29;185(7):1181-94
references.pmid: 19546244
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 28;103(48):18060-5
references.pmid: 17110448
references.citation: N Engl J Med. 2012 Apr 12;366(15):1423-32
references.pmid: 22494122
references.citation: Insect Mol Biol. 2011 Apr;20(2):215-24
references.pmid: 21114562
references.citation: Eur J Biochem. 2002 Feb;269(3):954-60
references.pmid: 11846797
references.citation: Am J Trop Med Hyg. 2007 Apr;76(4):659-65
references.pmid: 17426166
references.citation: Lancet. 2012 Feb 4;379(9814):413-31
references.pmid: 22305225
references.citation: Science. 1986 Mar 7;231(4742):1157-9
references.pmid: 3080809
references.citation: Adv Exp Med Biol. 2008;627:126-40
references.pmid: 18510020
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 19;108(16):6415-9
references.pmid: 21451136
references.citation: J Biol Chem. 1976 Sep 25;251(18):5551-7
references.pmid: 965377
references.citation: Anal Biochem. 1993 Apr;210(1):179-87
references.pmid: 8489015
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 23;105(38):14447-52
references.pmid: 18794531
references.citation: J Biochem. 2000 May;127(5):901-8
references.pmid: 10788801
references.citation: Curr Biol. 2002 Apr 30;12(9):712-23
references.pmid: 12007414
references.citation: Am J Trop Med Hyg. 2009 Oct;81(4):622-30
references.pmid: 19815877
references.citation: F1000 Biol Rep. 2010 Aug 19;2:62
references.pmid: 21173851
references.citation: Virol J. 2010 Sep 07;7:214
references.pmid: 20819232
references.citation: PLoS Pathog. 2012;8(4):e1002588
references.pmid: 22496640
references.citation: Microsc Res Tech. 2011 Nov;74(11):1032-9
references.pmid: 21509905
references.citation: J Proteomics. 2010 Oct 10;73(11):2064-77
references.pmid: 20685252
references.citation: Proc Natl Acad Sci U S A. 2004 Jan 13;101(2):470-5
references.pmid: 14707262
references.citation: Mol Cell. 2010 Dec 22;40(6):893-904
references.pmid: 21172655