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In this project we will focus on identification, expression and functional analysis of tick peptides related toevolutionarily conserved calcitonins. In insects and vertebrates, these peptides control essential physiologicalfunctions such as water and ion balance, metabolism, feeding behaviour and reproduction. We will use the ticksIxodes ricinus and I. scapularis as model species, as the role of neuropeptides in these vectors of dangerouspathogens is poorly understood. To elucidate signalling pathways of tick calcitonin-like peptides (CTs), we will usebioinformatics, cDNA cloning, qRT-PCR, in situ hybridisation and immunohistochemical staining with newly developed antibodies against tick CTs. Predicted receptors for CTs will be functionally characterised and theirspatial and temporal expression in tick organs determined by qRT-PCR. Electrophysiological techniques and microscopic video analysis will be used to determine effects of CTs on the salivary glands, digestive tract andreproductive organs in vitro. Changes in production and activity of digestive enzymes in intestinal extracts will beanalysed by Western blots. We will investigate peptide-receptor signalling pathways by suppressing expression of CTs receptors by RNA interference (RNAi). A very important part of this project is detailed analysis of effects of vaccination or CT receptor suppression by RNAi on transmission of pathogens Borrelia burgdorferi and B. afzelii. These modern and original approaches could reveal new mechanisms in tick blood feeding and reproduction, as well as very complex interactions between ticks, pathogens and hosts. |
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