BHLHB2 Controls Bdnf Promoter 4 Activity and Neuronal Excitability

Brain-derived neurotrophic factor (BDNF), via activation of TrkB receptors, mediates vital physiological functions in the brain, ranging from neuronal survival to synaptic plasticity, and has been implicated in the pathophysiology of neurodegenerative disorders. Although transcriptional regulation of the BDNF gene (Bdnf) has been extensively studied, much remains to be understood. We discovered a sequence within Bdnf promoter 4 that binds the basic helix-loop-helix protein BHLHB2 and is a target for BHLHB2-mediated transcriptional repression. NMDA receptor activation de-repressed promoter 4-mediated transcription and correlated with reduced occupancy of the promoter by BHLHB2 in cultured hippocampal neurons. Bhlhb2 gene −/− mice showed increased hippocampal exon 4-specific Bdnf mRNA levels compared with +/+ littermates under basal and activity-dependent conditions. Bhlhb2 knock-out mice also showed increased status epilepticus susceptibility, suggesting that BHLHB2 alters neuronal excitability. Together, these results support a role for BHLHB2 as a new modulator of Bdnf transcription and neuronal excitability.

[1]  A. Marini,et al.  Activity-dependent Release of Brain-derived Neurotrophic Factor Underlies the Neuroprotective Effect ofN-Methyl-d-aspartate* , 1998, The Journal of Biological Chemistry.

[2]  B. Dias,et al.  Differential regulation of multiple brain-derived neurotrophic factor transcripts in the postnatal and adult rat hippocampus during development, and in response to kainate administration. , 2004, Brain research. Molecular brain research.

[3]  H. Thoenen.  Neurotrophins and Neuronal Plasticity , 1995, Science.

[4]  H. Scharfman,et al.  BDNF and epilepsy: too much of a good thing? , 2001, Trends in Neurosciences.

[5]  Hilmar Bading,et al.  Nuclear calcium signaling controls CREB-mediated gene expression triggered by synaptic activity , 2001, Nature Neuroscience.

[6]  P. Anderson,et al.  Bcl-3 and NFκB p50-p50 Homodimers Act as Transcriptional Repressors in Tolerant CD4+ T Cells* , 2004, Journal of Biological Chemistry.

[7]  K. Magnusson.  Declines in mRNA Expression of Different Subunits May Account for Differential Effects of Aging on Agonist and Antagonist Binding to the NMDA Receptor , 2000, The Journal of Neuroscience.

[8]  M. Mattson,et al.  Telomerase Mediates the Cell Survival-Promoting Actions of Brain-Derived Neurotrophic Factor and Secreted Amyloid Precursor Protein in Developing Hippocampal Neurons , 2002, The Journal of Neuroscience.

[9]  A. Basbaum,et al.  Resistance to excitotoxin-induced seizures and neuronal death in mice lacking the preprotachykinin A gene. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Hyde,et al.  Detection of plasmid DNA vectors following gene transfer to the murine airways , 2005, Gene Therapy.

[11]  Marcelo A Wood,et al.  Transgenic mice expressing a truncated form of CREB-binding protein (CBP) exhibit deficits in hippocampal synaptic plasticity and memory storage. , 2005, Learning & memory.

[12]  S. Egan,et al.  Stra13 Homodimers Repress Transcription through Class B E-box Elements* , 2002, The Journal of Biological Chemistry.

[13]  H. Bading,et al.  The Yin and Yang of NMDA receptor signalling , 2003, Trends in Neurosciences.

[14]  G. Collingridge,et al.  Long-term potentiation of NMDA receptor-mediated synaptic transmission in the hippocampus , 1991, Nature.

[15]  A. Burny,et al.  Overlapping CRE and E Box Motifs in the Enhancer Sequences of the Bovine Leukemia Virus 5′ Long Terminal Repeat Are Critical for Basal and Acetylation-Dependent Transcriptional Activity of the Viral Promoter: Implications for Viral Latency , 2004, Journal of Virology.

[16]  C. Shatz,et al.  Synaptic Activity and the Construction of Cortical Circuits , 1996, Science.

[17]  Bing Li,et al.  Enhancement of Synaptic Plasticity through Chronically Reduced Ca2+ Flux during Uncorrelated Activity , 2004, Neuron.

[18]  Kaia Palm,et al.  Mouse and rat BDNF gene structure and expression revisited , 2006, Journal of neuroscience research.

[19]  Xueying Jiang,et al.  BDNF Variation and Mood Disorders: A Novel Functional Promoter Polymorphism and Val66Met are Associated with Anxiety but Have Opposing Effects , 2005, Neuropsychopharmacology.

[20]  I. Black,et al.  Trophic regulation of synaptic plasticity. , 1999, Journal of neurobiology.

[21]  M. Barton,et al.  The seizure-related phenotype of brain-derived neurotrophic factor knockdown mice , 2005, Neuroscience.

[22]  Y. Jan,et al.  Neuronal type information encoded in the basic-helix-loop-helix domain of proneural genes. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[23]  H. Bading,et al.  Opposing roles of synaptic and extrasynaptic NMDA receptors in neuronal calcium signalling and BDNF gene regulation , 2003, Current Opinion in Neurobiology.

[24]  P. Walker,et al.  Transcriptional Activation of the Human Brain-derived Neurotrophic Factor Gene Promoter III by Dopamine Signaling in NT2/N Neurons* , 2003, Journal of Biological Chemistry.

[25]  Eric C. Griffith,et al.  Derepression of BDNF Transcription Involves Calcium-Dependent Phosphorylation of MeCP2 , 2003, Science.

[26]  R. Taneja,et al.  Cross-regulatory interaction between Stra13 and USF results in functional antagonism , 2001, Oncogene.

[27]  N. Copeland,et al.  A highly efficient recombineering-based method for generating conditional knockout mutations. , 2003, Genome research.

[28]  Hong Sun,et al.  Stra13 expression is associated with growth arrest and represses transcription through histone deacetylase (HDAC)-dependent and HDAC-independent mechanisms. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  O. Ohara,et al.  Distinctive four promoters collectively direct expression of brain-derived neurotrophic factor gene. , 1994, Brain research. Molecular brain research.

[30]  K. Nave,et al.  SHARPs: mammalian enhancer-of-split- and hairy-related proteins coupled to neuronal stimulation. , 1997, Molecular and cellular neurosciences.

[31]  T. Bonhoeffer,et al.  Bidirectional Activity-Dependent Morphological Plasticity in Hippocampal Neurons , 2004, Neuron.

[32]  Mariza de Andrade,et al.  Human brain derived neurotrophic factor (BDNF) genes, splicing patterns, and assessments of associations with substance abuse and Parkinson's Disease , 2005, American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics.

[33]  R. Malenka,et al.  CREB modulates excitability of nucleus accumbens neurons , 2006, Nature Neuroscience.

[34]  L. Olson,et al.  Identification of cells in rat brain and peripheral tissues expressing mRNA for members of the nerve growth factor family , 1990, Neuron.

[35]  M. Mattson,et al.  Basic Fibroblast Growth Factor Selectively Increases AMPA‐Receptor Subunit GluR1 Protein Level and Differentially Modulates Ca2+ Responses to AMPA and NMDA in Hippocampal Neurons , 1995, Journal of neurochemistry.

[36]  E. Nestler,et al.  Histone Modifications at Gene Promoter Regions in Rat Hippocampus after Acute and Chronic Electroconvulsive Seizures , 2004, The Journal of Neuroscience.

[37]  M. Mattson,et al.  Glucagon‐like peptide‐1 decreases endogenous amyloid‐β peptide (Aβ) levels and protects hippocampal neurons from death induced by Aβ and iron , 2003 .

[38]  M. Einarson,et al.  Regulation of Id1 and its association with basic helix-loop-helix proteins during nerve growth factor-induced differentiation of PC12 cells , 1995, Molecular and cellular biology.

[39]  C. Culmsee,et al.  Roles of nuclear factor kappaB in neuronal survival and plasticity. , 2000, Journal of neurochemistry.

[40]  A. Routtenberg,et al.  Transcription factor NF‐κB activation after in vivo perforant path LTP in mouse hippocampus , 2004, Hippocampus.

[41]  H. Manev,et al.  NMDA-stimulated expression of BDNF mRNA in cultured cerebellar granule neurones. , 1993, Neuroreport.

[42]  Daisuke Hattori,et al.  DNA Methylation-Related Chromatin Remodeling in Activity-Dependent Bdnf Gene Regulation , 2003, Science.

[43]  B. Gwag,et al.  Activation of NMDA receptors increases brain-derived neurotrophic factor (BDNF) mRNA expression in the hippocampal formation. , 1993, Neuroreport.

[44]  Q. Yan,et al.  Expression of brain-derived neurotrophic factor protein in the adult rat central nervous system , 1997, Neuroscience.

[45]  C. Culmsee,et al.  Roles of Nuclear Factor κB in Neuronal Survival and Plasticity , 2000 .

[46]  Brian J. Wiltgen,et al.  NF-kappa B functions in synaptic signaling and behavior. , 2003, Nature neuroscience.

[47]  A. Romano,et al.  Participation of Rel/NF-κB transcription factors in long-term memory in the crab Chasmagnathus , 2000, Brain Research.

[48]  M E Greenberg,et al.  Requirement for BDNF in activity-dependent survival of cortical neurons. , 1994, Science.

[49]  E. Kandel,et al.  cAMP response element-binding protein is activated by Ca2+/calmodulin- as well as cAMP-dependent protein kinase. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[50]  O. Lindvall,et al.  Rapid increase of BDNF mRNA levels in cortical neurons following spreading depression: regulation by glutamatergic mechanisms independent of seizure activity. , 1993, Brain research. Molecular brain research.

[51]  R. Ricciardi,et al.  DNA Binding of Repressor Nuclear Factor-κB p50/p50 Depends on Phosphorylation of Ser337 by the Protein Kinase A Catalytic Subunit* , 2005, Journal of Biological Chemistry.

[52]  M. Mattson,et al.  Glucagon-like peptide-1 decreases endogenous amyloid-beta peptide (Abeta) levels and protects hippocampal neurons from death induced by Abeta and iron. , 2003, Journal of neuroscience research.

[53]  B. McEwen,et al.  The role of N‐methyl‐D‐asparate receptors in neurogenesis , 2006, Hippocampus.

[54]  Li I. Zhang,et al.  Heterosynaptic Scaling of Developing GABAergic Synapses: Dependence on Glutamatergic Input and Developmental Stage , 2007, The Journal of Neuroscience.

[55]  R. Flavell,et al.  Defective T cell activation and autoimmune disorder in Stra13-deficient mice , 2001, Nature Immunology.

[56]  David Baltimore,et al.  NF-κB functions in synaptic signaling and behavior , 2003, Nature Neuroscience.

[57]  A. Roopra,et al.  The Basic Helix-Loop-Helix Protein, SHARP-1, Represses Transcription by a Histone Deacetylase-dependent and Histone Deacetylase-independent Mechanism* , 2001, The Journal of Biological Chemistry.

[58]  B. Lu,et al.  Role of Neurotrophins in Synapse Development and Plasticity , 1997, Reviews in the neurosciences.

[59]  A. Harris,et al.  The hypoxia‐regulated transcription factor DEC1 (Stra13, SHARP‐2) and its expression in human tissues and tumours , 2004, The Journal of pathology.

[60]  A. Wellstein,et al.  Mitogen-induced Expression of the Fibroblast Growth Factor-binding Protein Is Transcriptionally Repressed through a Non-canonical E-box Element* , 2000, The Journal of Biological Chemistry.

[61]  W. Quinn,et al.  Induction of a dominant negative CREB transgene specifically blocks long-term memory in Drosophila , 1994, Cell.

[62]  T. Bliss,et al.  A synaptic model of memory: long-term potentiation in the hippocampus , 1993, Nature.

[63]  M. Chao,et al.  Neurotrophins and their receptors: A convergence point for many signalling pathways , 2003, Nature Reviews Neuroscience.

[64]  I. Black,et al.  Brain-derived neurotrophic factor rapidly enhances phosphorylation of the postsynaptic N-methyl-D-aspartate receptor subunit 1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[65]  Steven Finkbeiner,et al.  Ca2+ Influx Regulates BDNF Transcription by a CREB Family Transcription Factor-Dependent Mechanism , 1998, Neuron.

[66]  I. Black,et al.  BDNF acutely increases tyrosine phosphorylation of the NMDA receptor subunit 2B in cortical and hippocampal postsynaptic densities. , 1998, Brain research. Molecular brain research.

[67]  Robert Lipsky,et al.  The excitoprotective effect of N‐methyl‐d‐aspartate receptors is mediated by a brain‐derived neurotrophic factor autocrine loop in cultured hippocampal neurons , 2005, Journal of neurochemistry.

[68]  A. Konnerth,et al.  Neurotrophin-evoked rapid excitation through TrkB receptors , 1999, Nature.

[69]  S. Pomeroy,et al.  Abnormal Cerebellar Development and Foliation in BDNF −/− Mice Reveals a Role for Neurotrophins in CNS Patterning , 1997, Neuron.

[70]  Michael E. Greenberg,et al.  CREB: A mediator of long-term memory from mollusks to mammals , 1994, Cell.

[71]  I. Black,et al.  Brain-derived neurotrophic factor modulates hippocampal synaptic transmission by increasing N-methyl-D-aspartic acid receptor activity. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[72]  Wen G. Chen,et al.  Upstream Stimulatory Factors Are Mediators of Ca2+-Responsive Transcription in Neurons , 2003, The Journal of Neuroscience.

[73]  A. Novelli,et al.  Nuclear factor κB is a critical determinant in N‐methyl‐d‐aspartate receptor‐mediated neuroprotection , 2001, Journal of neurochemistry.

[74]  I. Black,et al.  Brain-derived neurotrophic factor acutely enhances tyrosine phosphorylation of the AMPA receptor subunit GluR1 via NMDA receptor-dependent mechanisms. , 2004, Brain research. Molecular brain research.

[75]  H. Phillips,et al.  Widespread expression of BDNF but not NT3 by target areas of basal forebrain cholinergic neurons. , 1990, Science.

[76]  Lawrence M. Grover,et al.  Two components of long-term potentiation induced by different patterns of afferent activation , 1990, Nature.

[77]  A. Marini,et al.  Role of brain-derived neurotrophic factor and NF-kappaB in neuronal plasticity and survival: From genes to phenotype. , 2004, Restorative neurology and neuroscience.

[78]  A. Routtenberg,et al.  Gene expression of the transcription factor NF-kappa B in hippocampus: regulation by synaptic activity. , 1996, Brain research. Molecular brain research.

[79]  F. Vajda.  Neuroprotection and neurodegenerative disease , 2002, Journal of Clinical Neuroscience.

[80]  M. Saarma,et al.  Multiple promoters direct tissue-specific expression of the rat BDNF gene , 1993, Neuron.

[81]  J. Tsien,et al.  c-fos regulates neuronal excitability and survival , 2002, Nature Genetics.