The purpose of this study was to determine whether the cytochemically defined distribution of sulfated macromolecules is significantly different in microdomains of basement membranes (BMs) associated with different levels of pulmonary airways. The high iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP) technique, which is highly specific for sulfate esters of glycosaminoglycans and some glycoproteins, was used as a probe to compare the BM of trachea, bronchi, and three different sizes of bronchioles. When HID-reactive sites were counted and statistically compared, significant differences were found between the three known anatomically distinct layers of the BM--lamina lucida (LL), lamina densa (LD), and lamina reticularis (LR)--relative to the airway level. The highest concentration of HID reactivity in trachea, bronchi, and large bronchiole was found in LR and the lowest in LD. By comparison, HID-reactive sites were found to be more concentrated in the LL in medium and small bronchioles. HID reactivity was consistently low in LD as compared with LL and LR in all five locations. The overall degree of HID reactivity in BMs was clearly highest in large bronchioles and lowest in medium and small bronchioles. This cytochemically detectable heterogeneity in the distribution of HID reactivity in BM microdomains may represent specific compositional differences in pulmonary BMs which are important determinants of epithelial cell function and might be expected to impact key biologic processes in normal and pathologic states.