Abstract
Clathrin heavy chain has been shown to be important for viability, embryogenesis, and RNA interference (RNAi) in arthropods such as Drosophila melanogaster. However, the functional roles of clathrin heavy chain in chelicerate arthropods, such as the predatory mite Metaseiulus occidentalis, remain unknown. We previously showed that dsRNA ingestion, followed by feeding on spider mites, induced systemic and robust RNAi in M. occidentalis females. In the current study, we performed a loss-of-function analysis of the clathrin heavy chain gene in M. occidentalis using RNAi. We showed that ingestion of clathrin heavy chain dsRNA by M. occidentalis females resulted in gene knockdown and reduced longevity. In addition, clathrin heavy chain dsRNA treatment almost completely abolished oviposition by M. occidentalis females and the few eggs produced did not hatch. Finally, we demonstrated that clathrin heavy chain gene knockdown in M. occidentalis females significantly reduced a subsequent RNAi response induced by ingestion of cathepsin L dsRNA. The last finding suggests that clathrin heavy chain may be involved in systemic RNAi responses mediated by orally delivered dsRNAs in M. occidentalis.
Highlights
Endocytosis, a crucial cellular process in all eukaryotes, is involved in the uptake of nutrients and control of the density of receptors on the cell surface [1,2,3]
Ablation of the clathrin heavy chain gene in the oocytes of Caenorhabditis elegans using RNA interference (RNAi) resulted in dead embryos and reduced yolk uptake [12]. These studies suggest that the highly conserved clathrin heavy chain protein plays an important role in the endocytotic process in many eukaryotes and is likely to be essential for viability and embryogenesis in multicellular organisms
We recently showed that orally delivered dsRNA induced robust and systemic RNAi responses in M. occidentalis females, but only after spider mite prey was provided [21]
Summary
Endocytosis, a crucial cellular process in all eukaryotes, is involved in the uptake of nutrients and control of the density of receptors on the cell surface [1,2,3]. Clathrin heavy chain gene knockdown by RNAi reduced subsequent RNAi responses in Drosophila cell cultures, possibly by blocking the uptake of dsRNA through clathrinmediated endocytosis (or receptor-mediated endocytosis) [13,14]. We hypothesized that clathrin heavy chain is involved in the RNAi responses in M. occidentalis following the ingestion of dsRNA.
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