Distribution of anionic sites during increasing tight junctional permeability in the rat submandibular gland

We sought to determine the effect of substance P salivary stimulation on both electrical charges and cellular permeability in the tight junctions of rat submandibular gland cells. Microperoxidase (1,900 daltons) was used as a tracer. It was administered by close-arterial infusion via the glandular arteries, and secretory routes of acinar cells in the gland were determined cytochemically. In the resting gland, microperoxidase reaction product filled the lateral intercellular spaces up to the tight junctions, but did not penetrate them. In the substance P-stimulated gland, microperoxidase reaction product was present within tight junctions and the lumen. Both distribution and mobility of anionic sites on the surface of the submandibular gland cells were studied utilizing multivalent ligand, ruthenium red and cationized ferritin as probes. In the resting gland, ruthenium red deposits were located uniformly in all areas of the basal membrane and intercellular spaces except for the tight junctional region of acinar and ductal cells. In substance P-stimulated gland, ruthenium red deposits were present in the tight junctional region and, to a lesser extent, in the intercellular spaces. Electrical charges of the tight junctions area of the lateral plasma membrane were studied using intraductal injection of cationized ferritin. In the resting gland, cationized ferritin probe was present in the intercellular spaces and was bound weakly in the tight junctional region. In the substance P-stimulated gland, cationized ferritin was firmly adherent to the tight junctional region.