PDE7 inhibitor TC3.6 ameliorates symptomatology in a model of primary progressive multiple sclerosis.

BACKGROUND AND PURPOSE: cAMP plays an important role in the transduction of signalling pathways involved in neuroprotection and immune regulation. Control of the levels of this nucleotide by inhibition of cAMP-specific PDEs such as PDE7 may affect the pathological processes of neuroinflammatory diseases like multiple sclerosis (MS). In the present study, we evaluated the therapeutic potential of the selective PDE7 inhibitor, TC3.6, in a model of primary progressive multiple sclerosis (PPMS), a rare and severe variant of MS. EXPERIMENTAL APPROACH: Theiler's murine encephalomyelitis virus-induced demyelinated disease (TMEV-IDD) is one of the models used to validate the therapeutic efficacy of new drugs in MS. As recent studies have analysed the effect of PDE7 inhibitors in the EAE model of MS, here the TMEV-IDD model was used to test their efficacy in a progressive variant of MS. Mice were subjected to two protocols of TC3.6 administration: on the pre-symptomatic phase and once the disease was established. KEY RESULTS: Treatment with TC3.6 ameliorated the disease course and improved motor deficits of infected mice. This was associated with down-regulation of microglial activation and reduced cellular infiltrates. Decreased expression of pro-inflammatory mediators such as COX-2 and the cytokines, IL-1ß, TNF-α, IFN-γ and IL-6 in the spinal cord of TMEV-infected mice was also observed after TC3.6 administration. CONCLUSION: These findings support the importance of PDE7 inhibitors, and specifically TC3.6, as a novel class of agents with therapeutic potential for PPMS. Preclinical studies are needed to determine whether their effects translate into durable clinical benefits.

Glycogen synthase kinase-3 inhibitors as potent therapeutic agents for the treatment of Parkinson disease.

Parkinson's disease (PD) is a devastating neurodegenerative disorder characterized by degeneration of the nigrostriatal dopaminergic pathway. Because the current therapies only lead to temporary, limited improvement and have severe side effects, new approaches to treat PD need to be developed. To discover new targets for potential therapeutic intervention, a chemical genetic approach involving the use of small molecules as pharmacological tools has been implemented. First, a screening of an in-house chemical library on a well-established cellular model of PD was done followed by a detailed pharmacological analysis of the hits. Here, we report the results found for the small heterocyclic derivative called SC001, which after different enzymatic assays was revealed to be a new glycogen synthase kinase-3 (GSK-3) inhibitor with IC(50) = 3.38 ± 0.08 µM. To confirm that GSK-3 could be a good target for PD, the evaluation of a set of structurally diverse GSK-3 inhibitors as neuroprotective agents for PD was performed. Results show that inhibitors of GSK-3 have neuroprotective effects in vitro representing a new pharmacological option for the disease-modifying treatment of PD. Furthermore, we show that SC001 is able to cross the blood-brain barrier, protects dopaminergic neurons, and reduces microglia activation in in vivo models of Parkinson disease, being a good candidate for further drug development.

Decreased CCAAT/enhancer binding protein ß expression inhibits the growth of glioblastoma cells.

C/EBPß is a leucine-zipper transcription factor implicated in the control of metabolism, development, cell differentiation, and proliferation. However, it remains unclear its role in tumor development. Here, we show that down-regulation of C/EBPß by RNA interference inhibits proliferation in the GL261 murine glioblastoma cell line, induces an arrest of the cell cycle at the G0/G1 boundary, and diminishes their transformation capacity and migration. In addition, we show that C/EBPß regulates the expression of several DNA damage response- and invasion-related genes. Lastly, C/EBPß depletion significantly retards tumor onset and prolongs survival in a murine orthotopic brain tumor model. Immunohistochemical analysis revealed a significant diminution of proliferating cell nuclear antigen (PCNA) labeling in tumors derived from C/EBPß-depleted GL261 cells compared with that in controls. These results show, for the first time, the dependence of glioma cells on C/EBPß and suggest a potential role of this transcription factor in glioma development.

Cancer stem cells and brain tumors.

Besides the role of normal stem cells in organogenesis, cancer stem cells are thought to be crucial for tumorigenesis. Most current research on human tumors is focused on molecular and cellular analysis of the bulk tumor mass. However, evidence in leukemia and, more recently, in solid tumors suggests that the tumor cell population is heterogeneous. In recent years, several groups have described the existence of a cancer stem cell population in different brain tumors. These neural cancer stem cells (NCSC) can be isolated by cell sorting of dissociated suspensions of tumor cells for the neural stem cell marker CD133. These CD133+ cells -which also express nestin, an intermediate filament that is another neural stem cell marker- represent a small fraction of the entire brain tumor population. The stem-like cancer cells appear to be solely responsible for propagating the disease in laboratory models. A promising new approach to treating glioblastoma proposes targeting cancer stem cells. Here, we summarize progress in delineating NCSC and the implications of the discovery of this cell population in human brain tumors.

Cancer stem cells and brain tumors

Besides the role of normal stem cells in organogenesis, cancer stem cells are thought to be crucial for tumorigenesis. Most current research on human tumors is focused on molecular and cellular analysis of the bulk tumor mass. However, evidence in leukemia and, more recently, in solid tumors suggests that the tumor cell population is heterogeneous. In recent years, several groups have described the existence of a cancer stem cell population in different brain tumors. These neural cancer stem cells (NCSC) can be isolated by cell sorting of dissociated suspensions of tumor cells for the neural stem cell marker CD133. These CD133+ cells -which also express nestin, an intermediate filament that is another neural stem cell marker- represent a small fraction of the entire brain tumor population. The stem-like cancer cells appear to be solely responsible for propagating the disease in laboratory models. A promising new approach to treating glioblastoma proposes targeting cancer stem cells. Here, we summarize progress in delineating NCSC and the implications of the discovery of this cell population in human brain tumors (AU)

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The mitochondrial respiratory complex I is a target for 15-deoxy-delta12,14-prostaglandin J2 action.

The prostaglandin J2 derivative 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) is a very active compound with important effects on inflammation, apoptosis, and cell growth processes. To exert this broad range of effects, 15d-PGJ2 binds and alters the activity of diverse proteins, which consequently are postulated to be mediators of its action. Among them are the transcription factors peroxisome proliferator-activated receptor gamma and nuclear factor kappaB, which are thought to play an essential role in the antitumorigenic and anti-inflammatory actions of 15d-PGJ2. Here, we show that 15d-PGJ2, at micromolar concentrations, efficiently blocks state 3 oxygen consumption in intact nonsynaptic mitochondria isolated from rat cerebral cortex. This effect is attributable to the inhibition by this prostaglandin of the activity of the enzyme NADH-ubiquinone reductase (complex I) of the mitochondrial respiratory chain. In addition to this, 15d-PGJ2 dramatically increases the rate of reactive oxygen species generation by complex I. The inhibition by 15d-PGJ2 of complex I activity was abolished by dithiothreitol, which raises the possibility that adduct formation with a critical component of complex I accounts for the inhibitory effect of this prostaglandin. These results clearly identified mitochondrial complex I as a new target for 15d-PGJ2 actions.

The peroxisome proliferator-activated receptor gamma is an inhibitor of ErbBs activity in human breast cancer cells.

One of the most interesting recent developments in the nuclear receptor field has been the identification of natural and synthetic agonists of the peroxisome proliferator-activated receptor (PPAR) family, coupled with a growing recognition that the gamma isoform (PPARgamma) affects pathways important in a variety of human diseases. Here we show that the activation of PPARgamma through the 15-deoxy-Delta-12,14-prostaglandin J(2) (PG-J(2)) ligand causes a dramatic inhibition of ErbB-2 and ErbB-3 tyrosine phosphorylation caused by neuregulin 1 (NRG1) and neuregulin 2 (NRG2) in MCF-7 cells. This effect is accompanied by a very efficient blocking of ErbBs effects upon proliferation, differentiation and cell death in these cells. Preincubation of MCF-7 cells with PG-J(2) before addition of NRG1 and NRG2 had a dramatic growth-suppressive effect accompanied by accumulation of cells in the G0/G1 compartment of the cell cycle, and a marked increase in apoptosis. NRG1 and NRG2 induce G1 progression, which was associated with stimulation of the phosphatidylinositol-3 kinase (PI 3-K) pathway, whereas survival was dependent on ERK1/ERK2 activation. Both pathways were inhibited by PG-J(2). Furthermore, PG-J(2) can abolish the NRG1 and NRG2-induced increase in anchorage-independent growth of these cells. PG-J(2) also blocks phosphorylation of other receptor tyrosine kinases, such as IGF-IR, in MCF-7 cells, and suppress proliferation of other breast cancer cell lines. In summary, our data show a specific inhibitory action of PG-J(2) on the activity of the ErbB receptors in breast cancer cells.

Thyroid hormone regulates oxidative phosphorylation in the cerebral cortex and striatum of neonatal rats.

We have previously shown that thyroid hormone (T(3)) regulates mitochondrial gene expression, morphology and transmembrane potential in the developing brain. Here, we have analysed the effect of thyroid hormone on mitochondrial function in different brain regions. For this purpose we have determined, in control, hypothyroid and T(3)-treated hypothyroid neonatal rats, the rate of oxidative phosphorylation in isolated mitochondria and the activity of the respiratory complexes in tissue homogenates. Our results showed a decrease in oxidative phosphorylation rate (only in the presence of NADH-generating substrates) and mitochondrial complexes I and III activity in the cerebral cortex and striatum of hypothyroid neonates, but not in the other areas analysed (hippocampus, cerebellum, thalamus, mid brain and brain stem). In parallel with mitochondrial activity, the levels of mitochondrially encoded transcripts were decreased only in the cerebral cortex and striatum of hypothyroid rats. The administration of T(3) corrected all these parameters. In summary, this study showed a down-regulation of mitochondrial gene expression accompanied by a decrease in mitochondrial activity in the cerebral cortex and striatum of developing hypothyroid neonatal rats.

Characterization of the promoter region of the rat CCAAT/enhancer-binding protein alpha gene and regulation by thyroid hormone in rat immortalized brown adipocytes.

CCAAT/enhancer binding proteins (C/EBP) are a family of transcription factors with a highly conserved basic/leucine zipper (bZIP) domain that has been implicated in the transcriptional control of genes involved in cell growth and differentiation. We have previously demonstrated that the expression of C/EBPalpha and C/EBPbeta genes is regulated by thyroid hormone in rat liver during development. The aim of the present study was to explore the molecular mechanisms underlying the control of C/EBPalpha gene expression by thyroid hormone. To achieve this goal, we isolated and characterized a genomic clone containing 1171 bp of the 5'-flanking region of the rat C/EBPalpha gene. This fragment was an active promoter in MB492 cells, an immortalized brown adipocyte cell line that expresses the endogenous C/EBPalpha gene in a T3-dependent manner. Sequence analysis suggested the presence of three thyroid hormone response elements, TRE-1 (-602/-589), TRE2 (-411/-396), and TRE3 (-376/-350). The results of deletion, mutagenesis, and gel mobility shift analysis disclosed that only TRE-1, an ER2-type response element, represented a functional T3 response element. Our results demonstrate that T3 is a factor that positively regulates C/EBPalpha gene expression in a direct fashion.

Involvement of the NGFI-A gene in the differentiation of neuroblastoma cells.

The transcription factor NGFI-A is an early response gene that has been implicated in the regulation of cell growth and differentiation and, more recently, in apoptosis. This gene is expressed in many tissues, and is very abundant in the brain. However, little is known about its functional role in the differentiation of this tissue. In the present work we investigated the role of NGFI-A in serum withdrawal-induced differentiation in N2A neuroblastoma cells. To do so, we studied the effect of NGFI-A antisense oligonucleotides and NGFI-A overexpression on this process. We show that neuroblastoma cells treated with an NGFI-A antisense oligonucleotide do not undergo normal morphological differentiation after serum withdrawal, whereas N2A cells overexpressing this gene extend long neurites, even in the presence of serum. We also show that NGFI-A overexpression is accompanied by an increase in the amount of phosphorylated microtubule-associated protein MAP1B, which has been associated with neurite outgrowth. Our results suggest that the NGFI-A gene plays an important role in neurite extension.

Control by thyroid hormone of NGFI-A gene expression in lung: regulation of NGFI-A promoter activity.

In previous studies, it was shown that the expression of the transcription factor NGFI-A is under thyroid hormone (T3) control in developing rat brain. Here, the effect of this hormone on the expression of this gene in lung, as well as the effect of T3 and NGFI-A protein on NGFI-A promoter activity, has been investigated. The results show that T3 regulates NGFI-A gene expression at a pretranslational level in neonatal and adult rat lung, but also at a translational and/or posttranslational level in 5-day-old neonates, since the NGFI-A protein content was decreased in hypothyroid animals without changes in mRNA. Transfection experiments showed an induction of NGFI-A promoter activity by thyroid hormone. NGFI-A protein blocked this stimulation, suggesting a negative interaction between NGFI-A protein and thyroid hormone receptor, which could explain the transient induction by T3 of this gene observed in the first 2-4 h.

Neuronal mitochondrial morphology and transmembrane potential are severely altered by hypothyroidism during rat brain development.

We recently demonstrated that thyroid hormone is an important regulator of mitochondrial gene expression during brain development. To gain further insights into the consequences of this regulation, we have performed functional and structural analysis of brain mitochondria from control and hypothyroid neonatal rats. Flow cytometric analysis showed a significant decrease in the mitochondrial transmembrane potential in hypothyroid animals compared with controls, which was reversed after 48 h, but not after 2 h, of thyroid hormone administration, suggesting that the functional alterations observed are the consequence of changes in mitochondrial gene expression. In addition, band shift studies showed a protein bound to the rat mitochondrial promoter differentially regulated by thyroid state. Electron microscopic analysis of cerebral cortex, striatum, and hippocampus revealed marked differences in the morphology of neuronal mitochondria from control and hypothyroid neonates. Hypothyroid mitochondria presented a decrease in the area of the inner membrane plus cristae in all areas studied, except for the hippocampal CA1 neurons and nonneuronal cell types. The observations reported here provide a basis for the known biochemical action of thyroid hormone on brain development.

Regulation by thyroid hormone and retinoic acid of the CCAAT/enhancer binding protein alpha and beta genes during liver development.

The effect of thyroid hormone and retinoic acid on the expression of CCAAT/enhancer binding proteins (C/EBPs) alpha and beta was investigated in rat liver during development. Congenital hypothyroidism caused a significant decrease in both C/EBP alpha and C/EBP beta gene expression at early stages of postnatal development. This effect was tissue specific since thyroid hormone had no effect on the level of C/EBP mRNAs in brown fat. Injection of 15-, and 30-day-old hypothyroid animals with thyroid hormone resulted in a slow recovery of the hepatic levels of both C/EBP alpha and beta mRNA levels. Retinoic acid was a very rapid and potent stimulator compared to thyroid hormone in hypothyroid animals. C/EBP alpha and beta protein levels were markedly diminished in hypothyroid neonates and the kinetics of induction of these proteins by thyroid hormone was faster than the one observed for the corresponding transcripts. The discrepancies observed between mRNA and protein levels suggest a translational or post-translational regulation of these genes as the major point of thyroid hormone action on these genes.

Thyroid hormone controls the expression of insulin-like growth factor I receptor gene at different levels in lung and heart of developing and adult rats.

Thyroid hormone exerts profound effects on the insulin-like growth factors (IGFs)/IGF factor I receptor (IGF-IR) system through its action on the production of IGF-I peptide and IGF-binding proteins. Most of these actions are mediated by the direct control of pituitary GH gene by thyroid hormone. In this work, we have analyzed the possible effect of hypothyroidism on the expression of IGF-IR gene, both in adult and developing animals. Our results show that in the lung and heart, thyroid hormone exerts a negative effect on IGF-IR gene expression in the adult animals and during perinatal life (from day 15 onwards). This negative effect is exerted at different levels. In the heart, this regulation occurs at a pretranslational level, indicated by the fact that parallel changes in the number of membrane IGF-I receptors and IGF-IR transcripts were observed, whereas in lung, no effect of thyroid hormone was noted in the amount of IGF-IR transcripts, suggesting a translational or posttranslational control. GH does not seem to mediate T3 effects on this gene. In contrast, retinoic acid increases the expression of IGF-IR gene at a transcriptional or posttranscriptional level in adult lung and heart. Because the IGFE/ IGF-IR system is depressed in hypothyroid animals, the specific increase in the number of IGF-IRs in the lung and heart of these animals could represent a mechanism to ameliorate the negative effects of hypothyroidism on these important organs.

Effect of perinatal hypothyroidism on the developmental regulation of rat pituitary growth hormone and thyrotropin genes.

We studied the effects of thyroid hormone (T3) on GH, TSH, and T3 receptor (TR) gene expression as well as deiodinase activities during rat pituitary development. By reverse transcriptase-polymerase chain reaction, GH and TSH beta transcripts were detectable on fetal day 15. Although with certain differences, the expression of both GH and TSH beta genes was under T3 control during fetal and neonatal life. Differences in plasma, but not pituitary, TSH concentrations were observed between control and hypothyroid animals throughout the period studied. Both TR alpha and TR beta genes were expressed in the fetal pituitary. TR alpha 1, TR beta 2, and c-erbA alpha 2 transcripts displayed a developmental profile different from that of TR beta 1. Thyroid hormone repressed TR alpha 1, TR beta 2, and c-erbA alpha 2 and stimulated TR beta 1. Type I and type II deiodinase activities (5'DI and 5'DII, respectively) had different ontogenic patterns; 5'D-II was the predominant activity in fetuses, with levels similar to those in adults, whereas the level of 5'D-I was low and increased with age. T3 stimulated 5'D-I and decreases 5'D-II. These results demonstrate that in somatotroph and thyrotroph cells, the mechanisms responsible for T3 action are mature and active very early in development and suggest an involvement of this hormone in the establishment and/or maintenance of the somatotroph and thyrotroph phenotype.

Thyroid hormone-regulated brain mitochondrial genes revealed by differential cDNA cloning.

Thyroid hormone (T3) plays a critical role in the development of the central nervous system and its deficiency during the early neonatal period results in severe brain damage. However the mechanisms involved and the genes specifically regulated by T3 during brain development are largely unknown. By using a subtractive hybridization technique we have isolated a number of cDNAs that represented mitochondrial genes (12S and 16S rRNAs and cytochrome c oxidase subunit III). The steady state level of all three RNAs was reduced in hypothyroid animals during the postnatal period and T3 administration restored control levels. During fetal life the level of 16S rRNA was decreased in the brain of hypothyroid animals, suggesting a prenatal effect of thyroid hormone on brain development. Since T3 does not affect the amount of mitochondrial DNA, the results suggest that the effect of T3 is at transcriptional and/or postranscriptional level. In addition, the transcript levels for two nuclear-encoded mitochondrial cytochrome c oxidase subunits: subunits IV and VIc were also decreased in the brains of hypothyroid animals. Hypothyroidism-induced changes in mitochondrial RNAs were followed by a concomitant 40% decrease in cytochrome c oxidase activity. This study shows that T3 is an important regulator of mitochondrial function in the neonatal brain and, more importantly, provides a molecular basis for the specific action of this hormone in the developing brain.

Induction of NGFI-B gene expression during T cell activation. Role of protein phosphatases.

mRNA expression of the immediate-early gene NGFI-B was investigated in T cells during the G0/G1 transition as well as throughout the G1 phase. After stimulation of T lymphocytes with Con A or phorbol 12,13 dibutyrate (PDBu), NGFI-B gene expression showed an induction of at least sevenfold within 3 h of stimulation. Twenty-four h later, however, the level of NGFI-B transcripts had fallen to almost basal levels. Activation of the Ca2+ signaling pathway also produced an induction of this gene, although to a lesser extent than the one obtained after protein kinase C activation. Similar transient kinetics of NGFI-B mRNA were also observed after PDBu stimulation of G1 lymphoblasts. However, the induction of NGFI-B by IL-2 is dependent on the presence of cycloheximide. Con A-induced activation of NGFI-B gene expression was not overcome by cyclosporin A or by 8Br-cAMP, but was partially prevented by dexamethasone. In lymphoblasts, okadaic acid caused the induction of NGFI-B gene expression, indicating a role for the serine/threonine protein phosphatases PP1 and PP2A in the regulation of this gene in resting cells. Okadaic acid-induced NGFI-B transcripts were significantly more stable than PDBu-induced NGFI-B mRNA. Thus, the level of NGFI-B transcripts in T cells might be determined by the balance between the activities of several serine/threonine protein kinases and phosphatases. Together, these findings indicate that the transient induction of NGFI-B transcripts is associated with normal lymphocyte activation. Because the mRNA for NGFI-B codes for a zinc-finger DNA-binding protein, these results suggest that NGFI-B participates in transcriptional regulation during T cell activation.

Sequence of the penicillin-binding protein 2-encoding gene (penA) of Neisseria perflava/sicca.

The nucleotide sequence of the transpeptidase domain (Tp) of the penicillin (Pn)-binding protein 2-encoding gene (penA) from two Pn-sensitive (PnS) and two Pn-resistant (PnR) strains of Neisseria perflava/sicca, has been determined. The sequences from PnS strains, although displaying high homology with those PnS strains from other Neisseria species, also showed several interesting modifications suggesting that these isolates might represent intermediates in the mutational development of PnR in N. perflava/sicca.

Differential effect of thyroid hormone on NGFI-A gene expression in developing rat brain.

NGFI-A is an immediate early gene that is rapidly activated in quiescent cells by mitogens or in postmitotic neurons after depolarization. We have previously shown that the expression of NGFI-A in the developing rat brain is under the control of thyroid hormone. Now we report, by means of in situ hybridization histochemistry, the differential effect of thyroid hormone on NGFI-A expression in distinct brain regions depending on the developmental stage. NGFI-A messenger RNA (mRNA) content was analyzed in the piriform cortex, striatum, hippocampus, and cerebral cortex of control, hypothyroid, and T3-injected hypothyroid rats at birth and on postnatal days 5 and 15. In the newborn rats, experimental hypothyroidism is associated with reduced levels of NGFI-A mRNA in most of the brain regions studied. On postnatal day 15, the difference in NGFI-A expression between control and hypothyroid rats is less apparent in the striatum or no longer present in the piriform cortex and the hippocampus. In the cerebral cortex, hypothyroidism is associated with reduced levels of NGFI-A mRNA on postnatal day 15. The dentate gyrus is always insensitive to the thyroidal state. Administration of T3 accelerates the recovery of NGFI-A mRNA in 5- and 15-day-old rats. However, in newborn rats, the effect of the hormone is noticeable only in the piriform cortex. We also show that the reduced level of NGFI-A mRNA in hypothyroidism is accompanied by a reduction in the protein level. Convulsions induced by pentylenetetrazole administration resulted in an increased expression of the NGFI-A gene, which is of similar magnitude in control and hypothyroid rats.

Perinatal hypothyroidism impairs the normal transition of GLUT4 and GLUT1 glucose transporters from fetal to neonatal levels in heart and brown adipose tissue. Evidence for tissue-specific regulation of GLUT4 expression by thyroid hormone.

GLUT1 and GLUT4 glucose transporter expression is highly regulated in muscle and adipose tissue during perinatal life. Here we have investigated the role of thyroid hormones in the regulation of GLUT4 induction and GLUT1 repression associated to neonatal development. Perinatal hypothyroidism markedly impaired GLUT4 protein induction in heart. This effect was heart specific, and a greater expression of GLUT4 was detected in brown adipose tissue from neonatal hypothyroid rats compared with controls. These changes in GLUT4 protein expression were not detected in brown adipose tissue or heart when hypothyroidism was induced in adult rats. These results indicate that GLUT4 induction during perinatal life is highly sensitive to thyroid hormones in both heart and adipose tissue. Perinatal hypothyroidism was characterized by decreased cardiac GLUT4 mRNA concentrations. T3 injection caused a marked increase in cardiac levels of GLUT4 mRNA in hypothyroid neonates. Thus, in 13-day-old hypothyroid rats, GLUT4 mRNA levels increased 3-fold 1 h after T3 injection. Under these conditions, retinoic acid also caused a rapid increase in cardiac GLUT4 mRNA levels from hypothyroid neonates. In addition, cardiac levels of GLUT4 protein markedly increased in fetuses and in neonates 24 h after T3 injection. These findings suggest that a direct effect of thyroid hormones is the promotion of cardiac GLUT4 gene expression. GLUT1 protein expression was markedly enhanced in brown adipose tissue and heart during neonatal hypothyroidism as well as in hypothyroidism induced in adult rats. This was concomitant to greater levels of GLUT1 mRNA in hearts from hypothyroid neonates. Immunofluorescence analysis indicated that cardiomyocytes from hypothyroid pups contained an enhanced level of GLUT1 protein. Furthermore, T3 injection caused a decrease in cardiac levels of GLUT1 mRNA in hypothyroid neonates. These results indicate that thyroid hormone manipulation leads to inverse regulation of GLUT1 and GLUT4 glucose transporter gene expression in the neonatal heart. We conclude that thyroid hormones play a pivotal role controlling the transition of glucose transporter carriers from fetal to neonatal levels in heart and brown adipose tissue.