Malaria is the major vector-borne parasitic disease for much of the developing world and also threatens military personnel, tourists, business people and others who work in Africa, Asia and Latin America. Because of parasite and mosquito resistance to drugs and pesticides, new control strategies are needed. The long-term objective of the research is to understand the physiology, biochemistry, and genetics of mosquitoes that are resistant to malaria infections. This may suggest ways to replace natural populations with genetically transformed nonvector mosquitoes. A specific type of resistance is found in a genetically- selected strain of Anopheles gambiae which melanotically encapsulates ookinetes of many malaria species. A fully susceptible strain was also selected for comparison. The proposed research will focus on examining the biochemical and genetic basis for resistance. A newly-discovered model system of resistance that uses ion exchange beads will be used for genetic and biochemical studies. Plasmodium- resistant mosquitoes melanize negatively charged C-25 beads while susceptible mosquitoes do not. This system will greatly facilitate quantitative trait loci mapping of resistance genes, since no parasites need to be introduced and males can be scored. Markers to be used for mapping include an esterase locus which has been shown to be associated with resistance and microsatellite markers. A total of 137 microsatellite markers are now available from all autosomes and the X-chromosome. In addition to these mapping studies, the biochemistry of melanization in resistant mosquitoes and the lack of melanization in susceptible mosquitoes will also be examined. C-25 beads inoculated into susceptible mosquitoes and then dissected and introduced into resistant mosquitoes are not melanized. The nature of this protection will be studied by treatment of the beads with various enzymes prior to inoculation into resistant mosquitoes. SDS PAGE will be used to examine proteins on several types of beads destined to be melanized in the resistant strain and on unmelanized beads in the susceptible strain. Proteins associated with melanization or with the protective response in susceptible mosquitoes will be purified and used for polyclonal antibody production.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI037083-01A1
Application #
2073682
Study Section
Special Emphasis Panel (ZRG5-TMP (01))
Project Start
1995-09-30
Project End
1998-07-31
Budget Start
1995-09-30
Budget End
1996-07-31
Support Year
1
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Zoology
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Lapcharoen, Parichat; Komalamisra, Narumon; Rongsriyam, Yupha et al. (2012) Investigations on the role of a lysozyme from the malaria vector Anopheles dirus during malaria parasite development. Dev Comp Immunol 36:104-11
Kajla, Mayur K; Shi, Lei; Li, Bin et al. (2011) A new role for an old antimicrobial: lysozyme c-1 can function to protect malaria parasites in Anopheles mosquitoes. PLoS One 6:e19649
Kajla, Mayur K; Andreeva, Olga; Gilbreath 3rd, Thomas M et al. (2010) Characterization of expression, activity and role in antibacterial immunity of Anopheles gambiae lysozyme c-1. Comp Biochem Physiol B Biochem Mol Biol 155:201-9
Paskewitz, Susan M; Andreev, Olga (2008) Silencing the genes for dopa decarboxylase or dopachrome conversion enzyme reduces melanization of foreign targets in Anopheles gambiae. Comp Biochem Physiol B Biochem Mol Biol 150:403-8
Paskewitz, S M; Li, B; Kajla, M K (2008) Cloning and molecular characterization of two invertebrate-type lysozymes from Anopheles gambiae. Insect Mol Biol 17:217-25
Li, Bin; Huang, Yu; Paskewitz, Susan M (2006) Hen egg white lysozyme as an inhibitor of mushroom tyrosinase. FEBS Lett 580:1877-82
Shi, L; Li, B; Paskewitz, S M (2006) Cloning and characterization of a putative inhibitor of melanization from Anopheles gambiae. Insect Mol Biol 15:313-20
Li, Bin; Paskewitz, Susan M (2006) A role for lysozyme in melanization of Sephadex beads in Anopheles gambiae. J Insect Physiol 52:936-42
Paskewitz, Susan M; Shi, Lei (2005) Bypassing the midgut results in development of Plasmodium berghei oocysts in a refractory strain of Anopheles gambiae (Diptera: Culicidae). J Med Entomol 42:712-5
Paskewitz, Susan M; Shi, Lei (2005) The hemolymph proteome of Anopheles gambiae. Insect Biochem Mol Biol 35:815-24

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