Properties of human atrial natriuretic factor (ANF) binding to the crude membrane fraction of rat kidney were studied using the ANF-radiolabeled receptor assay; the developmental change of renal ANF receptors in three age groups of spontaneously hypertensive rats (SHR) was also investigated with the methods of radiolabeled receptor assay and the quantitative approach of in vitro macro-autoradiography. Temperature and incubation time greatly influenced ANF binding capacities because of the degradation of radiolabeled ligand. Addition of 5 mM MgCl2 to assay buffer was useful for the stabilization of ANF specific binding. Scatchard analysis suggested that the crude membrane fraction of rat's kidney had a single binding site with the apparent dissociation constant of 0.55 nM. In the study of the developmental change of renal ANF receptor in SHR, systolic blood pressure of the SHR at the age of 5 weeks and 12 weeks was significantly higher than that of age-matched Wistar-Kyoto (WKY) rats, but there was no significant difference in blood pressure between SHR and WKY rats at the age of 3 weeks. Concerning the radiolabeled receptor assay of ANF, the apparent dissociation constant and maximum binding capacity in SHR were low in all age groups when compared with those of WKY rats. In the in vitro macro-autoradiographic observation, the specific binding of ANF was localized mainly in the renal cortex, and these binding patterns of SHR and WKY rats were the same in all age groups. The specific binding in the kidney of SHR was proved to be quantitatively smaller than that in WKY rats using the research analysis sytem (RAS 1,000) as investigated in radiolabeled receptor assay. These findings suggest that the kidneys of SHR have a higher affinity and lower capacity binding sites (possibly genetic in origin) for ANF from a young age, and these changes in the renal ANF receptor may affect the progression of the hypertensive state in SHR.
ASJC Scopus subject areas
- Internal Medicine