Article Figures & Data
Figures
- FIG. 1.
Schematic diagram of Acanthocheilonema viteae chitinase gene sequences. Identified genes with lettered exons are shown with filled boxes. Blank boxes represent introns, while intergenic regions are shown as lines.
- FIG. 2.
Southern blot analysis of Acanthocheilonema viteae genomic DNA. DNAs from Meriones jirds (M) and A. viteae (A) (20 μg per lane) were digested with PvuII (P), HindIII (H), and EcoRI (E). Digested DNAs in ethidium bromide-stained gels are shown on the left, while hybridization results for both samples are shown on the right.
- FIG. 3.
Quantification of chitinase transcripts in different Acanthocheilonema viteae stages by real-time PCR. The ordinate depicts the log10 values of relative chitinase expression, calculated by the 2−ΔΔCT method. The values represent the relative amounts of chitinase normalized to the endogenous reference (tropomyosin) and relative to the calibrator (chitinase) from the late male L4, since it had the lowest level of chitinase expression of all life stages.
- FIG. 4.
Western blot analysis of expression of chitinase protein in different stages of Acanthocheilonema viteae. Under reducing conditions (50 mM dithiothreitol), 12 μg of A. viteae male (M) and female (F) worms, blood microfilariae (Mf), uterine microfilariae (ut Mf), and L3 and L4 homogenates of 42 larvae were resolved by 12% SDS-PAGE and transferred to nitrocellulose. Blots were probed with mouse anti-A. viteae chitinase serum.
- FIG. 5.
Reduction of release of microfilariae from female worms following RNAi of Acanthocheilonema viteae chitinase gene I. AvChi dsRNA, a total of 30 adult worms (six groups of five worms each) were treated with chitinase dsRNA; Mal dsRNA, 30 adult worms (six groups of five worms each) were treated with dsRNA for the control maltose binding protein. Asterisks represent significant differences between AvChi dsRNA- and Mal dsRNA-treated groups (P < 0.01).
Tables
- TABLE 1.
Phenotypic changes and decrease in chitinase transcripts after chitinase RNAi treatment of different A. viteae stagesa
Parasite stage Phenotypic change(s) Influence of RNAi on chitinase transcript level L3 87% inhibition of molting to L4 93% decrease Adult female 50% mortality, 57 to 68% decrease in release of microfilariae, 42 to 58% inhibition of hatching of microfilariae 18 to 20% decrease in chitinase transcripts of adult female worms, 85% decrease in chitinase transcripts of uterine contents Adult male No changes No decrease ↵ a Values depict averages for three independent experiments.
