Hexose-6-phosphate dehydrogenase and 11beta-hydroxysteroid dehydrogenase-1 tissue distribution in the rat.
Intracellular concentrations of the glucocorticoids cortisol and corticosterone are modulated by the enzymes 11beta-hydroxysteroid dehydrogenase (11beta-HSD) 1 and 2. 11beta-HSD1 is a reduced nicotinamide adenine dinucleotide phosphate (NADPH)-dependent microsomal reductase that converts the inactive glucocorticoids cortisone and 11-dehydrocorticosterone to their active forms, cortisol and corticosterone. Hexose-6-phosphate dehydrogenase (H6PDH) is an enzyme that generates NADPH from oxidized NADP (NADP(+)) within the endoplasmic reticulum. In the absence of NADPH or H6PDH to regenerate NADPH, 11beta-HSD1 acts as a dehydrogenase and inactivates glucocorticoids, as does 11beta-HSD2. A monoclonal antibody against H6PDH was produced to study the possibility that 11beta-HSD1 in the absence of H6PDH may be responsible for hydroxysteroid dehydrogenase activity in tissues that do not express significant amounts of 11beta-HSD2. H6PDH and 11beta-HSD1 expression was surveyed in a variety of rat tissues by real-time RT-PCR, Western blot analysis, and immunohistochemistry. H6PDH was found in a wide variety of tissues, with the greatest concentrations in the liver, kidney, and Leydig cells. Although the brain as a whole did not express significant amounts of H6PDH, some neurons were clearly immunoreactive by immunohistochemistry. H6PDH was amply expressed in most tissues examined in which 11beta-HSD1 was also expressed, with the notable exception of the renal interstitial cells, in which dehydrogenase activity by 11beta-HSD1 probably moderates activation of the glucocorticoid receptor because rat renal interstitial cells do not have significant amounts of mineralocorticoid receptors. This antibody against the H6PDH should prove useful for further studies of enzyme activity requiring NADPH generation within the endoplasmic reticulum.
Gomez-Sanchez
Elise P
EP
Research Service, GV (Sonny) Montgomery Veterans Affairs Medical Center, Jackson, MS 39216, USA. egomez-sanchez@medicine.umsmed.edu
Romero
Damian G
DG
de Rodriguez
Angela F
AF
Warden
Mary P
MP
Krozowski
Zygmunt
Z
Gomez-Sanchez
Celso E
CE
D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002237:Carbohydrate Dehydrogenases / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D007150:Immunohistochemistry; D007668:Kidney / Q000201:enzymology; D007985:Leydig Cells / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008297:Male; D009928:Organ Specificity; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002237:Carbohydrate Dehydrogenases; C020552:galactose-6-phosphate dehydrogenase; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1