Browsing by Author "Keating, Joseph"

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    Differential Induction of Proteins in Anopheles gambiae sensu stricto (Diptera: Cullicidae) Larvae in Response to Heavy Metal Selection
    (National Institute of Health, 2010-07-21) Mireji, Paul O.; Keating, Joseph; Kenya, Eucharia U.; Mbogo, Charles; Nyambaka, Hudson; Osir, Ellie; Githure, John; Beier, John
    Investigations were conducted to establish the magnitude and pattern of differential expression of proteins due to generational selection of third instar An. gambiae s.s. larvae by cadmium, copper and lead heavy metals, three possible common urban pollutants. A susceptible strain of An. gambiae s.s. third instar larvae was separately placed under selection pressure with cadmium, copper and lead at LC30 and controls through five generations. First, third and fifth generation selection survivors were screened for differentially expressed proteins relative to non-exposed control by two-dimensional gel electrophoresis. Distribution patterns of the spots were analysed by Chi Square or Fishers exact test and variations in expressions between and within generation by ANOVA. Most differentially expressed spots were acidic and of low molecular weight among all metals and generations. Type of heavy metals and generation were main indicators of variations in differential expressions. Variation between generations was most significant among cadmium-selected populations of which most number of spots were induced in the fifth generation. Most spots were induced in the copper-selected population in the third generation. The induced protein spots may be products from respective genes that respond to heavy metals and counter their toxicity, thus building An. gambiae s.s. tolerance to these pollutants. The differential pattern and magnitude of expressed spots has potential application as molecular markers for assessment of anopheline adaptation status to heavy metals, and provide insight into the extent of environmental pollution.
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    Expression of metallothionein and tubulin in heavy metal-tolerant Anopheles gambiae sensu stricto (Diptera:Culicidae).
    (Elsevier, 2010) Mireji, Paul O.; Keating, Joseph; Hassaali, Ahmed; Impoinvil, Daniel E.; Mbogo, Charles; Muturi, Martha; Nyambaka, Hudson; Kenya, Eucharia U.; Githure, John; Beier, John
    Anopheles mosquitoes have been shown to adapt to heavy metals in their natural habitats. In this study we explored the possibility of using Anopheles gambiae sensu stricto as bioreporters for environmental heavy metal pollution through expressions of their metalresponsive metallothionein and alpha-tubulin genes. The study was undertaken with third instar larvae after selection by cadmium, copper, or lead at LC(30) through five successive generations. Expression levels were determined in the 5th generation by semiquantitative RT-PCR on the experimental and control populations. The data were analyzed using one-way ANOVA. The highest metallothionein (F(3,11)=4.574, P=0.038) and alpha-tubulin (F(3,11)=12.961, P=0.002) responses were observed in cadmiumtolerant treatments. There was significantly higher expression of metallothionein in cadmium or copper treatments relative to the control (P=0.012), and in cadmium than in lead treatments (P=0.044). Expressions of alpha-tubulin were significantly higher in cadmium than in control treatments (P=0.008). These results demonstrate the capacity of An. gambiae s.s. to develop tolerance to increased levels of heavy metal challenge. The results also confirm the potential of heavy metal-responsive genes in mosquitoes as possible bio-indicators of heavy metal environmental pollution. How the tolerance and expressions relate to An. gambiae s.s. fitness and vectorial capacity in the environment remains to be elucidated.