Direct visualization of renin-cell distribution in preglomerular vascular trees dissected from rat kidney

Abstract

Three methods to visualize directly the distribution of granulated renin-positive cells in vascular trees microdissected from rat kidney were developed. Kidneys were removed from anesthesized rats, hemisectioned, macerated in HCl, and soaked in distilled water for 24-48 h. Cortical preglomerular vascular trees consisting of arcuate and cortical radial arteries and afferent arterioles were microdissected with the aid of a stereomicroscope. Granulated cells can be visualized in three ways. First, under transmitted or incident light observation, granulated cells are readily distinguished from the surrounding smooth muscle cells, because of marked differences in the refractive properties of these two cell types. Second, quinacrine, a fluorescent, intravital stain selective for dense-core granules, can be administered (2 mg/kg iv) to the rat 1 h before nephrectomy. When illuminated with 440-nm light, granulated cells fluorescence strongly at 510 nm. Third, specific immunostaining for renin can be obtained with a polyclonal anti-rat renin antibody and avidin-biotin immunoperoxidase staining in vascular trees subjected to cell permeabilization with Triton. These new techniques permit the direct visualization of the distribution of granulated renin-positive cells in preglomerular vessels under conditions in which the vascular architecture is largely preserved.