Postnatal development of carbonic anhydrase IV expression in rabbit kidney.

Abstract

Carbonic anhydrase (CA) IV activity facilitates renal acidification by catalyzing the dehydration of luminal carbonic acid. CA IV has been localized to the proximal tubules and medullary collecting ducts. Maturation of CA IV expression has been considered to be important in the development of renal acid excretion. The purpose of the present study was to determine the maturational expression of CA IV in rabbit kidney. A guinea pig polyclonal antibody to purified rabbit lung microsomal membrane CA IV was generated. Immunoblotting of membrane proteins after peptide-N-glycosidase F treatment revealed two N-glycosylation sites and reduction in size from approximately 52 to 35 kDa; there appeared to be heavier glycosylation in the medulla. In membrane and total proteins from the kidney cortex, CA IV was 15-30% of the adult level during the first 2 wk of life but increased to mature levels by 5 wk of age. The maturational pattern in the cortex was confirmed by measuring SDS-resistant CA hydratase activity. In the medulla, both membrane and total proteins were generally less than one-fourth of the adult level of CA IV during the first 2 wk of life before reaching mature levels by 5 wk of age. Immunohistochemistry showed staining in proximal tubules (apical > basolateral), with maximal label in the S2 segment. CA IV also appeared on the apical membranes of a minority cell type of the cortical collecting duct, presumably the alpha-intercalated cell. Several labeled cells also appeared to be the process of being extruded from medullary collecting ducts of 1- to 2-wk rabbits. The antibody did not reliably detect medullary CA IV expression in sections from mature rabbits. These studies indicate that there is a substantial postnatal increase in expression of CA IV in the maturing kidney in both the cortex and medulla. The disappearance of intercalated cells in the maturing rabbit medullary collecting duct may be part of a normal renal developmental program as previously reported [J. Kim, J.-H. Cha, C. C. Tisher, and K. M. Madsen. Am. J. Physiol. 270 (Renal Fluid Electrolyte Physiol. 39): F575-F592, 1996]. It is likely that the maturation of CA IV expression contributes to the increase in renal acidification observed early in postnatal life.