title: Effect of glucocorticoids on the activity, expression and proximal promoter of type II deiodinase in rat brown adipocytes.
abstract: Triiodothyronine (T3) is important for thermogenesis in brown adipose tissue (BAT). Type II deiodinase (DIO2) produces T3 required for intracellular needs in BAT. Brown adipocytes in culture require T3 for the adrenergic stimulation of DIO2. Glucocorticoids induce adipocyte differentiation (lipogenesis). We investigated the regulation of DIO2 activity, Dio2 mRNA and Dio2 promoter activity by glucocorticoids in primary cultures of rat brown adipocytes using dexamethasone (DEX) and hydrocortisone (HC). DEX and HC regulated the adrenergic stimulation of DIO2 activity in a dose- and time-dependent manner, inhibiting DIO2 activity at short treatment times and large doses (1-10 μM) and stimulating DIO2 at low HC doses (1-100 nM) and longer times (DEX). Insulin depletion reduced DIO2 activity but the response to glucocorticoids remained unchanged. DEX and HC inhibited basal DIO2 activity. DEX had no effect on DIO2 half-life, whereas HC stabilized DIO2 activity. DEX and HC inhibited the adrenergic stimulation of Dio2 mRNA expression (100-10000 nM, 14-96 h), but stabilized Dio2 mRNA, particularly DEX. DEX increased basal Dio2 mRNA levels, possibly through stabilization of Dio2 mRNA. An 807 bp construct of the murine Dio2 proximal promoter showed maximal reporter activity, with the cAMP response element (CRE) essential for transcriptional activity. DEX caused inhibition in most constructs containing the CRE element whereas HC stimulated reporter activity in the 807 bp construct. Glucocorticoids inhibited the adrenergic stimulation of Dio2 at the transcriptional level in brown adipocytes, although DIO2 activity increased with HC, possibly due to stabilization of Dio2 activity and mRNA. The CRE and cEBP elements of the Dio2 promoter seem involved in the regulation by glucocorticoids.
authors.affiliation: Molecular Physiopathology and Nervous System, Inst Investigaciones Biomedicas (IIB), Centro Mixto CSIC-UAM (Consejo Superior Investigaciones Cientificas and Universidad Autonoma Madrid), Madrid, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: Martinez-deMena
authors.affiliation: Molecular Physiopathology and Nervous System, Inst Investigaciones Biomedicas (IIB), Centro Mixto CSIC-UAM (Consejo Superior Investigaciones Cientificas and Universidad Autonoma Madrid), Madrid, Spain.
authors.forename: Rosa-Maria
authors.initials: RM
authors.lastname: Calvo
authors.affiliation: Molecular Physiopathology and Nervous System, Inst Investigaciones Biomedicas (IIB), Centro Mixto CSIC-UAM (Consejo Superior Investigaciones Cientificas and Universidad Autonoma Madrid), Madrid, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Garcia
authors.affiliation: Molecular Physiopathology and Nervous System, Inst Investigaciones Biomedicas (IIB), Centro Mixto CSIC-UAM (Consejo Superior Investigaciones Cientificas and Universidad Autonoma Madrid), Madrid, Spain. Electronic address: mjobregon@iib.uam.es.
authors.forename: Maria Jesus
authors.initials: MJ
authors.lastname: Obregon
mesh_terms: D052437:Adipocytes, Brown / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D005938:Glucocorticoids / Q000494:pharmacology*; D006207:Half-Life; D006854:Hydrocortisone / Q000494:pharmacology; D007453:Iodide Peroxidase / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D000071256:Uncoupling Protein 1 / Q000378:metabolism
chemical_list: D005938:Glucocorticoids; D012333:RNA, Messenger; C000604373:Ucp1 protein, rat; D000071256:Uncoupling Protein 1; D003907:Dexamethasone; D007453:Iodide Peroxidase; D006854:Hydrocortisone
keywords: Brown adipocytes; Dexamethasone; Hydrocortisone; Type 2 deiodinase DIO2