title: Excitotoxicity downregulates TrkB.FL signaling and upregulates the neuroprotective truncated TrkB receptors in cultured hippocampal and striatal neurons.
abstract: Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival through activation of TrkB receptors. The trkB gene encodes a full-length receptor tyrosine kinase (TrkB.FL) and its truncated (T1/T2) isoforms. We investigated the changes in TrkB protein levels and signaling activity under excitotoxic conditions, which are characteristic of brain ischemia, traumatic brain injury, and neurodegenerative disorders. Excitotoxic stimulation of cultured rat hippocampal or striatal neurons downregulated TrkB.FL and upregulated a truncated form of the receptor (TrkB.T). Downregulation of TrkB.FL was mediated by calpains, whereas the increase in TrkB.T protein levels required transcription and translation activities. Downregulation of TrkB.FL receptors in hippocampal neurons correlated with a decrease in BDNF-induced activation of the Ras/ERK and PLCγ pathways. However, calpain inhibition, which prevents TrkB.FL degradation, did not preclude the decrease in signaling activity of these receptors. On the other hand, incubation with anisomycin, to prevent the upregulation of TrkB.T, protected to a large extent the TrkB.FL signaling activity, suggesting that truncated receptors may act as dominant-negatives. The upregulation of TrkB.T under excitotoxic conditions was correlated with an increase in BDNF-induced inhibition of RhoA, a mediator of excitotoxic neuronal death. BDNF fully protected hippocampal neurons transduced with TrkB.T when present during excitotoxic stimulation with glutamate, in contrast with the partial protection observed in cells overexpressing TrkB.FL or expressing GFP. These results indicate that BDNF protects hippocampal neurons by two distinct mechanisms: through the neurotrophic effects of TrkB.FL receptors and by activation of TrkB.T receptors coupled to inhibition of the excitotoxic signaling.
authors.affiliation: CNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal.
authors.forename: João R
authors.initials: JR
authors.lastname: Gomes
authors.forename: João T
authors.initials: JT
authors.lastname: Costa
authors.forename: Carlos V
authors.initials: CV
authors.lastname: Melo
authors.forename: Federico
authors.initials: F
authors.lastname: Felizzi
authors.forename: Patrícia
authors.initials: P
authors.lastname: Monteiro
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Pinto
authors.forename: Ana R
authors.initials: AR
authors.lastname: Inácio
authors.forename: Tadeusz
authors.initials: T
authors.lastname: Wieloch
authors.forename: Ramiro D
authors.initials: RD
authors.lastname: Almeida
authors.forename: Mário
authors.initials: M
authors.lastname: Grãos
authors.forename: Carlos B
authors.initials: CB
authors.lastname: Duarte
mesh_terms: D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D019208:Brain-Derived Neurotrophic Factor / Q000502:physiology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000502:physiology; D016923:Cell Death / Q000187:drug effects* / Q000235:genetics; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism*; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation / Q000187:drug effects*; D018698:Glutamic Acid / Q000633:toxicity; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D007608:Kainic Acid / Q000633:toxicity*; D009474:Neurons / Q000378:metabolism; D018696:Neuroprotective Agents / Q000378:metabolism; D061251:Primary Cell Culture; D051381:Rats; D017208:Rats, Wistar; D020813:Receptor, trkB / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000502:physiology
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D007527:Isoenzymes; D018696:Neuroprotective Agents; D018698:Glutamic Acid; D000841:Anisomycin; D020813:Receptor, trkB; D002154:Calpain; D020742:rhoA GTP-Binding Protein; D007608:Kainic Acid