Visualization of Acetylcholine in the Mouse Brain by a Combination of Immunohistochemistry with Ionic Fixation

概要

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Although acetylcholine (Ach) is an important neurotransmitter, there is no adequate histochemical method to detect Ach in tissue sections. We therefore developed a method to visualize the localization of Ach in tissue sections by a combination of immunohistochemistry with ionic fixation. Brains of mice, perfused with 4% formaldehyde and 5% phosphomolybdic acid (PMA) in 0.1 M phosphate buffer (pH 7.4) under sodium pentobarbital anesthesia, were removed and immersed quickly in boiling PMA solution for 5 min. Frozen sections were cut and soaked in a buffer containing 5% PMA at 4°C for 30 min, and immersed in a buffer containing bovine serum albumin at 4°C for 1 hr. Then the sections were incubated with anti-Ach antibody at 4°C for 12 hr. Subsequently, the immunoreaction due to Ach was visualized by the peroxidase-antiperoxidase method. Nerve cells in the brain sections from the interpeduncular nucleus of the cerebrum and bipolar cells in the multiform layer of the cerebral cortex were immmunohistochemically stained with the anti-Ach antibody.
日本組織細胞化学会の論文
1995-06-01