Table 1.
Adult survival following larval targeting of dsx.
Fig 1.
Developmental silencing of dsx decreases adult female appendage size.
With respect to adult females fed control siRNA nanoparticles as larvae, mean adult wing areas (A), as well as mean adult wing (B), proboscis (C), antenna (D), and maxillary palp (E) lengths are significantly decreased in females fed dsx-KD A or dsx-KD B siRNA nanoparticles as larvae. Means for each control or experimental group are reported, and these were determined through the combination of data gathered in at least four biological replicate experiments. Error bars represent standard deviations. *** = P < 0.001.
Fig 2.
Decreased female fertility and fecundity is observed following developmental silencing of dsx.
Although the percentages of dsx-KD A, dsx-KD B, and control siRNA-microinjected adult females that mated (A) or took blood meals (B) did not significantly differ, adult females that were injected with dsx-KD A or dsx-KD B siRNA as pupae had decreased fecundity (C) and fertility (D). Means for each control or experimental group are reported, and these were determined through the combination of data gathered in at least three biological replicate experiments. Error bars represent standard deviations.** = P < 0.01; *** = P < 0.001.
Fig 3.
Multiple ovary phenotypes are observed following developmental silencing of dsx.
Adult females that were injected with dsx-KD A or dsx-KD B siRNA as pupae had decreased ovary length both pre- (A) and post-blood meals (B). Representative ovaries from control-, dsx-KD A, or dsx-KD B stained with phalloidin (red) and the nuclear marker TO-PRO-3 (blue) are shown below each graph in A and B. In comparison to control microinjected females, the number of ovarioles/ovary (E) and length of each ovariole (F) post-blood meal was significantly less in females microinjected with dsx-KD A and dsx-KD B siRNA. Means for each control or experimental group are reported, and these were determined through the combination of data gathered in at least three biological replicate experiments. Error bars represent standard deviations. *** = P < 0.001.
Fig 4.
Sensillar defects result from silencing of dsx.
Antennal sTrichodea (inset in C) are found on the adult female A. aegypti antenna (control-injected animal shown in A), while sBasiconica (inset in D) are observed on the adult female maxillary palp (control-injected animal shown in B). With respect to control-injected adult females, adult females that were injected with dsx-KD A or dsx-KD B siRNA as pupae had significantly shorter antennal sTrichodea (C) and maxillary palp sBasiconica (D). Mean sensillar lengths for each control or experimental condition are reported, and these were determined through the combination of data gathered in at least two biological replicate experiments. Error bars represent standard deviations. *** = P < 0.001.
Fig 5.
Terminal segments of adult female antennae oriented distal upward that were hybridized with OR 62 (A, B, C), OR 123 (D, E, F), OR 2 (G, H, I), or OR 9 (J, K, L) riboprobes (green, at left in each panel) are shown. Antennae were counter-stained with anti-Tubulin (red, center of each panel) and TO-PRO-3 nuclear stain (blue in overlays at right in each panel). OR 62 (A), OR 123 (D), OR 2 (G), and OR 9 transcripts are detected in ORNs of control-injected adult female antennae (A, D, G, J). Little expression of these transcripts could be detected in adult females injected with dsx-KD A (B, E, H, K) or dsx-KD B (C, F, I, J) siRNA as pupae. Representative results from a total of 24 antennae stained in two separate replicate experiments are shown, and analysis of OR expression intensity levels is presented in S3 Table. Scale bars = 25 microns. Arrowheads mark ORNs.
Fig 6.
Adult female lifespan decreases following developmental silencing of dsx.
Survivorship curves for animals injected as pupae with control, dsx-KD A, or dsx-KD B siRNA are shown. Analysis of data combined from three replicate experiments indicated that in comparison to the control, life span was significantly decreased in individuals injected with dsx-KD A (P<0.001) or dsx-KD B (P<0.001) siRNA.