Effect of Ixodes ricinus bites in skin gene expression [skin]
Tick bites often promote local allergic reactions in the skin. The mechanisms involved in this process is not fully understood. Here we investigated the local changes in the skin induced by Ixodes ricinus bites. Overall design: Ixodes ricinus were collected on vegetation in forested areas. After shaving the back of C3H/HeN mouse, a plastic cap was fixed with wax to the mouse's back. Five female ticks were introduced into the cap which was sealed. For the control animals, plastic cap was just fixed to the back of the animal. After 4 days of blood meal, the cap was removed, and all ticks were collected. At day 4, 10 and 14 days, the skin at the site of the tick bite were removed. For the control animals, the cap was removed, and skin biopsy were collected as for the animals bitten by ticks. Samples were stored in RNAlater. Total skin RNA was isolated with RNeasy Mini Kit (Qiagen), followed by cDNA synthesis using the SuperScript VILO cDNA Synthesis Kit (ThermoFisher Scientific). The Ion AmpliSeq Transcriptome Mouse Gene Expression Kit was used to prepare bar-coded libraries and sequenced by using an Ion S5 next-generation sequencer. The AmpliSeqRNA plug-in (ThermoFisher Scientific) was used to calculate differential gene expression analysis