TY - JOUR
T1 - Quantitation and cellular localization of 11β-HSD1 expression in murine thymus
AU - Nuotio-Antar, Alli M.
AU - Hasty, Alyssa H.
AU - Kovacs, William J.
N1 - Funding Information:
These studies were supported in part by a Vanderbilt University Discovery Grant (to WJK). WJK received support from the Diana and Richard C. Strauss Professorship in Biomedical Research at the University of Texas Southwestern Medical Center. AMN-A received support from NIH training grant T32 HD007043. The Vanderbilt Diabetes Research and Training Center (DK020593) provided essential core facilities. Anti-11β-HSD1 antibody was the generous gift of Dr. Zygmunt Krozowski. The K8 (TROMA-I) hybridoma antibody developed by Drs. Philippe Brulet and Rolf Kemler was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.
PY - 2006/5
Y1 - 2006/5
N2 - 11β-Hydroxysteroid dehydrogenase type I (11β-HSD1), an NADPH-dependent reductase, functions in intact cells to convert inactive 11-keto metabolites of glucocorticoids into biologically active glucocorticoids. The enzyme is thus capable of amplifying glucocorticoid action in tissues in which it is expressed. In the experiments presented here, we show that 11β-HSD1 is expressed in the murine thymus and that expression increases from late fetal development to maximal levels in the adult thymus. Quantitative real time-PCR, immunoblots, and assays of enzymatic activity reveal adult thymic expression of 11β-HSD1 mRNA and protein at levels approximately 6-7% of those observed in liver. Immunofluorescence experiments show that the enzyme is expressed in the medullary thymocytes and thymocytes present at the corticomedullary junction. These experiments extend our recognition of 11β-HSD1 expression in cells of the immune system and lend support to the notion that glucocorticoid signaling and amplification of those signals by regeneration of active glucocorticoids from inactive 11-keto metabolites might impact intrathymic T cell development and the establishment of the immune repertoire.
AB - 11β-Hydroxysteroid dehydrogenase type I (11β-HSD1), an NADPH-dependent reductase, functions in intact cells to convert inactive 11-keto metabolites of glucocorticoids into biologically active glucocorticoids. The enzyme is thus capable of amplifying glucocorticoid action in tissues in which it is expressed. In the experiments presented here, we show that 11β-HSD1 is expressed in the murine thymus and that expression increases from late fetal development to maximal levels in the adult thymus. Quantitative real time-PCR, immunoblots, and assays of enzymatic activity reveal adult thymic expression of 11β-HSD1 mRNA and protein at levels approximately 6-7% of those observed in liver. Immunofluorescence experiments show that the enzyme is expressed in the medullary thymocytes and thymocytes present at the corticomedullary junction. These experiments extend our recognition of 11β-HSD1 expression in cells of the immune system and lend support to the notion that glucocorticoid signaling and amplification of those signals by regeneration of active glucocorticoids from inactive 11-keto metabolites might impact intrathymic T cell development and the establishment of the immune repertoire.
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U2 - 10.1016/j.jsbmb.2006.01.011
DO - 10.1016/j.jsbmb.2006.01.011
M3 - Article
C2 - 16621520
AN - SCOPUS:33646182383
SN - 0960-0760
VL - 99
SP - 93
EP - 99
JO - Journal of Steroid Biochemistry and Molecular Biology
JF - Journal of Steroid Biochemistry and Molecular Biology
IS - 2-3
ER -
