*2.12. Mast Cells Evaluation*

Twenty-one days after reserpine injection brain and sciatic nerve were harvested. Tissues were fixed in 10% buffered formalin, and embedded in paraffin blocks. Seven-μm sections were prepared from paraffin-embedded tissues. After deparaffinization, sections were stained with toluidine blue in order to assess mast cell infiltration. The mast cells count was performed on each slide through a Leica DM6 (Milan, Italy) microscope.

## *2.13. TNF-*α *and IL-1*β *ELISA*

The quantifications of TNF-α and IL-1β were assayed following the instructions provided by R&D Systems Quantikine Rat TNF- α and IL-1β immunoassay kit [49].
