Brain processing
Animals were placed under deep anesthesia and perfused
with phosphate-buffered para-formaldehyde 4%, (pH 7.4)
fixative. The brains were rapidly removed and stored in
para-formaldehyde 4%, (pH 7.4) overnight (4 C). Finally,
brains were maintained in cryopreservative solution at
20 C until use. For immunohistochemical assessment,
brains were cut coronally in 40-lm-thick sections with a
vibratome (Leica).