Anti-Obesity Sodium Tungstate Treatment Triggers Axonal and Glial Plasticity in Hypothalamic Feeding Centers

Objective This study aims at exploring the effects of sodium tungstate treatment on hypothalamic plasticity, which is known to have an important role in the control of energy metabolism. Methods Adult lean and high-fat diet-induced obese mice were orally treated with sodium tungstate. Arcuate and paraventricular nuclei and lateral hypothalamus were separated and subjected to proteomic analysis by DIGE and mass spectrometry. Immunohistochemistry and in vivo magnetic resonance imaging were also performed. Results Sodium tungstate treatment reduced body weight gain, food intake, and blood glucose and triglyceride levels. These effects were associated with transcriptional and functional changes in the hypothalamus. Proteomic analysis revealed that sodium tungstate modified the expression levels of proteins involved in cell morphology, axonal growth, and tissue remodeling, such as actin, CRMP2 and neurofilaments, and of proteins related to energy metabolism. Moreover, immunohistochemistry studies confirmed results for some targets and further revealed tungstate-dependent regulation of SNAP25 and HPC-1 proteins, suggesting an effect on synaptogenesis as well. Functional test for cell activity based on c-fos-positive cell counting also suggested that sodium tungstate modified hypothalamic basal activity. Finally, in vivo magnetic resonance imaging showed that tungstate treatment can affect neuronal organization in the hypothalamus. Conclusions Altogether, these results suggest that sodium tungstate regulates proteins involved in axonal and glial plasticity. The fact that sodium tungstate could modulate hypothalamic plasticity and networks in adulthood makes it a possible and interesting therapeutic strategy not only for obesity management, but also for other neurodegenerative illnesses like Alzheimer’s disease.

[1]  M. Inagaki,et al.  Role of phosphorylation on the structural dynamics and function of types III and IV intermediate filaments. , 2007, Experimental cell research.

[2]  C. Beaulieu,et al.  The basis of anisotropic water diffusion in the nervous system – a technical review , 2002, NMR in biomedicine.

[3]  Yoji Kawano,et al.  Phosphorylation by Rho Kinase Regulates CRMP-2 Activity in Growth Cones , 2005, Molecular and Cellular Biology.

[4]  T. Lutz,et al.  Blunted Fasting-Induced Hypothalamic Activation and Refeeding Hyperphagia in Late-Onset Obesity , 2009, Neuroendocrinology.

[5]  A. Bortolozzi,et al.  Sodium tungstate regulates food intake and body weight through activation of the hypothalamic leptin pathway , 2011, Diabetes, obesity & metabolism.

[6]  J. Delgado-García,et al.  Molecular Bases of Caloric Restriction Regulation of Neuronal Synaptic Plasticity , 2008, Molecular Neurobiology.

[7]  S. Woods,et al.  Central nervous system control of food intake , 2000, Nature.

[8]  M. W. Schwartz,et al.  Central nervous system control of food intake and body weight , 2006, Nature.

[9]  T. Horvath,et al.  UCP2 mediates ghrelin’s action on NPY/AgRP neurons by lowering free radicals , 2009, Nature.

[10]  N. Kassis,et al.  A Dietary Fat Excess Alters Metabolic and Neuroendocrine Responses Before the Onset of Metabolic Diseases , 2009, Cellular and Molecular Neurobiology.

[11]  R. Gomis,et al.  Target identification of the novel antiobesity agent tungstate in adipose tissue from obese rats , 2005, Proteomics.

[12]  Roy J Martin,et al.  Dynamics of recovery of body composition after overfeeding, food restriction or starvation of mature female rats. , 1986, The Journal of nutrition.

[13]  R. Gomis,et al.  Tungstate is an effective antidiabetic agent in streptozotocin-induced diabetic rats: a long-term study , 2001, Diabetologia.

[14]  A. Shevchenko,et al.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. , 1996, Analytical chemistry.

[15]  R. Norgren,et al.  Expression of Fos during sham sucrose intake in rats with central gustatory lesions. , 2008, American journal of physiology. Regulatory, integrative and comparative physiology.

[16]  S. Bouret,et al.  Formation of Projection Pathways from the Arcuate Nucleus of the Hypothalamus to Hypothalamic Regions Implicated in the Neural Control of Feeding Behavior in Mice , 2004, The Journal of Neuroscience.

[17]  Khader M Hasan,et al.  Retrospective measurement of the diffusion tensor eigenvalues from diffusion anisotropy and mean diffusivity in DTI , 2006, Magnetic resonance in medicine.

[18]  S. Bouret,et al.  Hypothalamic neural projections are permanently disrupted in diet-induced obese rats. , 2008, Cell metabolism.

[19]  A. G. Roseberry,et al.  Rapid Rewiring of Arcuate Nucleus Feeding Circuits by Leptin , 2004, Science.

[20]  P. Basser,et al.  Microstructural and physiological features of tissues elucidated by quantitative-diffusion-tensor MRI. , 1996, Journal of magnetic resonance. Series B.

[21]  D. Hochstrasser,et al.  Information transfer between large and small two‐dimensional polyacrylamide gel electrophoresis , 1999, Electrophoresis.

[22]  K. Mikoshiba,et al.  Phosphorylation of CRMP2 (Collapsin Response Mediator Protein 2) Is Involved in Proper Dendritic Field Organization , 2012, The Journal of Neuroscience.

[23]  S. Bouret,et al.  Trophic Action of Leptin on Hypothalamic Neurons That Regulate Feeding , 2004, Science.

[24]  Sheng-Kwei Song,et al.  Reduced Axonopathy and Enhanced Remyelination After Chronic Demyelination in Fibroblast Growth Factor 2 (Fgf2)-Null Mice: Differential Detection With Diffusion Tensor Imaging , 2011, Journal of neuropathology and experimental neurology.

[25]  P. Basser,et al.  Toward a quantitative assessment of diffusion anisotropy , 1996, Magnetic resonance in medicine.

[26]  Y. Ihara,et al.  Neurofibrillary tangle-associated collapsin response mediator protein-2 (CRMP-2) is highly phosphorylated on Thr-509, Ser-518, and Ser-522. , 2000, Biochemistry.

[27]  M. Black,et al.  Newly assembled microtubules are concentrated in the proximal and distal regions of growing axons , 1992, The Journal of cell biology.

[28]  J. Flier,et al.  The Fat-Brain Axis Enters a New Dimension , 2004, Science.

[29]  J. Rodríguez-Gil,et al.  Insulin-like actions of tungstate in diabetic rats. Normalization of hepatic glucose metabolism. , 1994, The Journal of biological chemistry.

[30]  R. Coppari,et al.  A treasure trove of hypothalamic neurocircuitries governing body weight homeostasis. , 2010, Endocrinology.

[31]  S. Kalko,et al.  Integration of DIGE and Bioinformatics Analyses Reveals a Role of the Antiobesity Agent Tungstate in Redox and Energy Homeostasis Pathways in Brown Adipose Tissue *S , 2008, Molecular & Cellular Proteomics.

[32]  K. Kaibuchi,et al.  CRMP-2 induces axons in cultured hippocampal neurons , 2001, Nature Neuroscience.

[33]  Veeranna,et al.  Cdk5 and MAPK are associated with complexes of cytoskeletal proteins in rat brain. , 2000, Brain research. Molecular brain research.

[34]  R. Gomis,et al.  Tungstate decreases weight gain and adiposity in obese rats through increased thermogenesis and lipid oxidation. , 2005, Endocrinology.

[35]  R Gomis,et al.  Dual effects of sodium tungstate on adipocyte biology: inhibition of adipogenesis and stimulation of cellular oxygen consumption , 2009, International Journal of Obesity.

[36]  M. Gresser,et al.  Inhibition of phosphatase and sulfatase by transition-state analogues. , 1988, Biochemistry.

[37]  B. Oldfield,et al.  The endogenous actions of hypothalamic peptides on brown adipose tissue thermogenesis in the rat. , 2010, Endocrinology.

[38]  Kozo Kaibuchi,et al.  [Neuronal polarity]. , 2008, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[39]  T. Horvath,et al.  Brain circuits regulating energy homeostasis , 2008, Regulatory Peptides.

[40]  C. Servis,et al.  Proteomic analysis of cytokine induced proteins in human intestinal epithelial cells: Implications for inflammatory bowel diseases , 2002, Proteomics.

[41]  Jong Hoon Lee,et al.  Axial asymmetry of water diffusion in brain white matter , 2005, Magnetic resonance in medicine.

[42]  M. Cowley,et al.  Diet-induced obesity causes severe but reversible leptin resistance in arcuate melanocortin neurons. , 2007, Cell metabolism.

[43]  R. Marouga,et al.  The development of the DIGE system: 2D fluorescence difference gel analysis technology , 2005, Analytical and bioanalytical chemistry.

[44]  Eduardo Fernández,et al.  Proton magnetic resonance spectroscopy (1H-MRS) reveals the presence of elevated myo-inositol in the occipital cortex of blind subjects , 2009, NeuroImage.

[45]  A. Bortolozzi,et al.  A functional leptin system is essential for sodium tungstate antiobesity action. , 2009, Endocrinology.

[46]  M. Huentelman,et al.  Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem. , 2002, Journal of neurophysiology.

[47]  R. Cone.  Anatomy and regulation of the central melanocortin system , 2005, Nature Neuroscience.

[48]  R M Levenson,et al.  Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment II , 2006, Histopathology.

[49]  M. Dietrich,et al.  Feeding signals and brain circuitry , 2009, The European journal of neuroscience.

[50]  Richard Robitaille,et al.  Glial cells in synaptic plasticity , 2006, Journal of Physiology-Paris.

[51]  C. Tanaka,et al.  In vivo assessment of peripheral nerve regeneration by diffusion tensor imaging , 2011, Journal of magnetic resonance imaging : JMRI.

[52]  M. Muñoz,et al.  Effects of tungstate, a new potential oral antidiabetic agent, in Zucker diabetic fatty rats. , 2001, Diabetes.

[53]  M. Adams,et al.  Tungsten in biological systems. , 1996, FEMS microbiology reviews.

[54]  J. Backer,et al.  Activation of hypothalamic S6 kinase mediates diet-induced hepatic insulin resistance in rats. , 2008, The Journal of clinical investigation.

[55]  K. Kaibuchi,et al.  Ras regulates neuronal polarity via the PI3-kinase/Akt/GSK-3beta/CRMP-2 pathway. , 2006, Biochemical and biophysical research communications.

[56]  K. Kaibuchi,et al.  GSK-3β Regulates Phosphorylation of CRMP-2 and Neuronal Polarity , 2005, Cell.

[57]  K. Kaibuchi,et al.  CRMP-2 binds to tubulin heterodimers to promote microtubule assembly , 2002, Nature Cell Biology.

[58]  H. Berthoud,et al.  Neural systems controlling the drive to eat: mind versus metabolism. , 2008, Physiology.

[59]  R. Gomis,et al.  Phosphorylation events implicating p38 and PI3K mediate tungstate-effects in MIN6 beta cells. , 2007, Biochemical and biophysical research communications.

[60]  J. Honnorat,et al.  Collapsin response mediator proteins (CRMPs) , 2003, Molecular Neurobiology.

[61]  Jesús Avila,et al.  Sodium tungstate decreases the phosphorylation of tau through GSK3 inactivation , 2006, Journal of neuroscience research.

[62]  I. Alafuzoff,et al.  Glycosylation changes in Alzheimer’s disease as revealed by a proteomic approach , 2004, Neuroscience Letters.