Shahid Karim, kshahid@uri.edu, Bronwyn Kenny, bken6554@postoffice.uri.edu, Sivakamasundari Pichu, au-sudha@mail.uri.edu, Nathan J. Miller, natem@uri.edu, and Thomas N. Mather, tmather@uri.edu. University of Rhode Island, Center for Vector-Borne Disease, 9 East Alumni Ave, 324 Woodward Hall, Kingston, RI
Protein trafficking in secretory and endocytic pathways relies on precisely choreographed membrane events requiring the faithful pairing of cognate soluble N-ethylmaleimide–sensitive attachment protein receptors (SNAREs) between membranes. Intracellular pathogen trafficking may use similar processes. We investigated the functional role of two SNARE proteins, YKT6 (synaptobrevin homolog) and Syntaxin in blacklegged tick (Ixodes scapularis) salivary glands using both in vivo and in vitro RNA interference. Salivary YKT6 and Syntaxin genes were silenced in individual experiments by injecting adult ticks with 500 ng of dsRNA complementing each gene sequence. Disrupting expression of either YKT6 or Syntaxin by RNAi reduced the ability of ticks to feed successfully, as demonstrated by reduced engorgement weights, partial to complete feeding inhibition and inability of ticks to reach repletion or lay eggs. Incubating isolated salivary glands with dsRNA also disrupted mRNA and protein expression. The functional role of SNARE proteins in secreting the intracellular bacterium, Anaplasma phagocytophilum from PGE2-stimulated salivary glands will be discussed.
Species 1: Acari Ixodidae
Ixodes scapularis (blacklegged tick, deer tick)
Keywords: Salivary glands, SNARE
