Immunohistochemical localization of the 11 beta-hydroxysteroid dehydrogenase type II enzyme in human kidney and placenta.
It has been proposed that the inactivation of glucocorticoids by the enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta HSD) is an obligatory step in the kidney, permitting binding of aldosterone to the mineralocorticoid receptor, and in the placenta, protecting the fetus from high circulating levels of maternal glucocorticoids. Both low and high affinity isoforms of 11 beta HSD are known to exist, with evidence accumulating that the former species (11 beta HSD1) does not fulfill criteria that would allow it to perform these physiological functions. We have recently cloned a high affinity isoform of the enzyme (11 beta HSD2) from a human kidney library and have shown this species to possess all of the characteristics predicted from whole cell studies. In the present study we have raised a polyclonal antibody (HUH23) to a synthetic peptide deduced from the carboxy-terminus of the protein. The immunopurified antibody recognized a single band at 41,000 daltons on Western blots of mammalian cells transfected with an expression plasmid containing 11 beta HSD2, slightly smaller than the predicted 44,140 daltons protein. A single band of identical size was also seen in blots of human kidney and placenta, suggesting post-translational processing of the enzyme. Immunohistochemical studies on frozen sections of human kidney showed strong 11 beta HSD2 immunoreactivity in the cortical distal convoluted tubules and collecting ducts. Strong staining was also observed in medullary tubules, which had the appearance of collecting ducts and the thick ascending limb of Henle's loop. Staining of medium intensity was observed in vascular smooth muscle cells. Epithelial cells of glomeruli showed weak but detectable reactivity with HUH23. In the placenta, HUH23 antibody immunoreactivity was restricted to syncytial trophoblast cells in which strong staining was observed. These results suggest that the 11 beta HSD2 enzyme colocalizes with the mineralocorticoid receptor in the distal nephron where it allows aldosterone to occupy its physiological receptor. Furthermore, 11 beta HSD2 is also ideally situated in the placenta to protect the fetus from high circulating levels of maternal glucocorticoids.
Krozowski
Z
Z
Laboratory of Molecular Hypertension, Baker Institute of Medical Research, Prahran, Australia.
MaGuire
J A
JA
Stein-Oakley
A N
AN
Dowling
J
J
Smith
R E
RE
Andrews
R K
RK
D042842:11-beta-Hydroxysteroid Dehydrogenases; D000595:Amino Acid Sequence; D000906:Antibodies; D015153:Blotting, Western; D002460:Cell Line; D004848:Epithelium / Q000201:enzymology; D005260:Female; D006801:Humans; D006913:Hydroxysteroid Dehydrogenases / Q000032:analysis* / Q000096:biosynthesis; D007150:Immunohistochemistry / Q000379:methods; D007527:Isoenzymes / Q000032:analysis*; D007668:Kidney / Q000166:cytology / Q000201:enzymology*; D007672:Kidney Cortex / Q000201:enzymology; D007678:Kidney Glomerulus / Q000201:enzymology; D007685:Kidney Tubules, Collecting / Q000201:enzymology; D008861:Microsomes / Q000201:enzymology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010920:Placenta / Q000166:cytology / Q000201:enzymology*; D011247:Pregnancy; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection; D014327:Trophoblasts / Q000201:enzymology
D000906:Antibodies; D007527:Isoenzymes; D010455:Peptides; D011994:Recombinant Proteins; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases