Characterisation of estrogenic 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity in the human brain.
Estrogens play a crucial role in multiple functions of the brain and the proper balance of inactive estrone and active estradiol-17beta might be very important for their cerebral effects. The interconversion of estrone and estradiol-17beta in target tissues is known to be catalysed by a number of human 17beta-hydroxysteroid dehydrogenase (17beta-HSD) isoforms. The present study shows that enzyme catalysed interconversion of estrone and estradiol-17beta occurs in the human temporal lobe. The oxidative cerebral pathway preferred estradiol-17beta to Delta(5)-androstenediol and testosterone, whereas the reductive pathway preferred dehydroepiandrosterone (DHEA) to Delta(4)-androstenedione and estrone. An allosteric Hill kinetic for NAD-dependent oxidation of estradiol-17beta was observed, whereas a typical Michaelis-Menten kinetic was shown for NADPH-dependent reduction of estrone. Investigations of the interconversion of estrogens in cerebral neocortex (CX) and subcortical white matter (SC) preparations of brain tissue from 12 women and 10 men revealed no sex-differences, but provide striking evidence for the presence of at least one oxidative membrane-associated 17beta-HSD and one cytosolic enzyme that catalyses both the reductive and the oxidative pathway. Membrane-associated oxidation of estradiol-17beta was shown to be significantly higher in CX than in SC (P<0.05), whereas the cytosolic enzyme activities were significantly higher in SC than in CX (P<0.0005). Finally, real-time RT-PCR analyses revealed that besides 17beta-HSD types 4 and 5 also the isozymes type 7, 8, 10 and 11 show substantial expression in the human temporal lobe. The characteristics of the isozymes lead us to the conclusion that cytosolic 17beta-HSD type 5 is the best candidate for the observed cytosolic enzyme activities, whereas the data gave no clear answer to the question, which enzyme is responsible for the membrane-associated oxidation of estradiol-17beta. In conclusion, the study strongly suggests that different cell types and different isozymes are involved in the cerebral interconversion of estrogens, which might play a pivotal role in maintaining the functions of the central nervous system.
Steckelbroeck
Stephan
S
Department of Pharmacology, University of Pennsylvania School of Medicine, 130C John Morgan Building, 3620 Hamilton Walk, Philadelphia, PA 19104-6084, USA. st_steckelbroek@hotmail.com
Watzka
Matthias
M
Reissinger
Annette
A
Wegener-Toper
Petra
P
Bidlingmaier
Frank
F
Bliesener
Niklaas
N
Hans
Volkmar H J
VH
Clusmann
Hans
H
Ludwig
Michael
M
Siekmann
Lothar
L
Klingmüller
Dietrich
D
D015067:17-Hydroxysteroid Dehydrogenases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D015114:Androstenediol / Q000032:analysis / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002648:Child; D002675:Child, Preschool; D003687:Dehydroepiandrosterone / Q000032:analysis / Q000378:metabolism; D004958:Estradiol / Q000032:analysis / Q000378:metabolism; D004970:Estrone / Q000032:analysis / Q000378:metabolism; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D013702:Temporal Lobe / Q000201:enzymology; D013739:Testosterone / Q000032:analysis / Q000378:metabolism
D007527:Isoenzymes; D012333:RNA, Messenger; D004970:Estrone; D013739:Testosterone; D003687:Dehydroepiandrosterone; D004958:Estradiol; D015114:Androstenediol; D015067:17-Hydroxysteroid Dehydrogenases