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Figure 2 | BMC Developmental Biology

Figure 2

From: Chitosan/siRNA nanoparticle targeting demonstrates a requirement for single-minded during larval and pupal olfactory system development of the vector mosquito Aedes aegypti

Figure 2

Expression and siRNA-nanoparticle mediated knockdown of sim in the larval olfactory system. sim is expressed in the antennae (A) and brain (A1) of wildtype L4 larvae. Two sim siRNAs, sim430(C, C1) and sim718(D, D1), were found to effectively knockdown (KD) sim in both L4 antennae (C, D) and brains (C1, D1) when delivered to A. aegypti larvae via chitosan nanoparticle feeding. Control siRNA feedings did not impact sim expression (antenna in B; brain in B1). In wildtype (A1) and control-fed animal (B1) L4 brains, clusters of cells expressing sim transcripts (red asterisks) are detected adjacent to the antennal lobe, the boundaries of which are marked by the yellow dotted circles in panels A1-D1. The overall reduced number of antennal sensory neurons targeting to the antennal lobes in sim knockdown animals does not appear to result from cell death, as evidenced by the lack of cleaved-caspase 3 staining in the L4 antennae of sim knockdown animals (E3,4; compare to wildtype and control-fed animals in E1,2; nuclei are marked by TO-PRO in E). Note that while no cleaved-caspase 3 was detected in the antennae shown, positively labeled cells were detected in other wild-type A. aegypti pupal tissues (not shown), suggesting that the reagent employed in these studies is effective in A. aegypti. Acetylated tublin staining (green, F) revealed normal antennal sensory neuron axon bundles in sim430 and sim718 knockdown animals (compare to wildtype and control-fed antennae in F1 and F2, respectively). The proximal ends of antennae are oriented upward in panels A-D, E, and F. Dorsal is oriented upward in panels A1-D1. Scale bars = 25 microns.

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