van Koppen_1988_J.Pharmacol.Exp.Ther_244_760

To visualize muscarinic receptors in human bronchi, the stripping film method was used which permits direct autoradiographic localization of tissue labeling. Cryostate sections of human bronchi were fixed in 0.5% glutaraldehyde in Krebs-Ringer buffer, pH 7.0 for 30 min at 0 degrees C, washed in Krebs-Ringer buffer for 20 min at 0 degrees C and incubated with (-)-[3H]Quinuclidinyl benzilate [(-)-[3H]QNB] for 90 min at 37 degrees C. Specific (-)-[3H]QNB binding to tissue sections was saturable (receptor density of 0.14 +/- 0.03 fmol/tissue section) and of high affinity (Kd of 40 +/- 9 pM). For autoradiography, labeled tissue sections were covered with stripping film and exposed for 5 months. Muscarinic receptors in human bronchi were located predominantly in submucosal glands and parasympathetic ganglia. There was less labeling in smooth muscle cells and nerve bundles. Epithelium and blood vessels located within the bronchial wall were devoid of specific labeling.