title: DNA methylation regulates neuronal glutamatergic synaptic scaling.
abstract: Enhanced receptiveness at all synapses on a neuron that receive glutamatergic input is called cell-wide synaptic upscaling. We hypothesize that this type of synaptic plasticity may be critical for long-term memory storage within cortical circuits, a process that may also depend on epigenetic mechanisms, such as covalent chemical modification of DNA. We found that DNA cytosine demethylation mediates multiplicative synaptic upscaling of glutamatergic synaptic strength in cultured cortical neurons. Inhibiting neuronal activity with tetrodotoxin (TTX) decreased the cytosine methylation of and increased the expression of genes encoding glutamate receptors and trafficking proteins, in turn increasing the amplitude but not frequency of miniature excitatory postsynaptic currents (mEPSCs), indicating synaptic upscaling rather than increased spontaneous activity. Inhibiting DNA methyltransferase (DNMT) activity, either by using the small-molecule inhibitor RG108 or by knocking down Dnmt1 and Dnmt3a, induced synaptic upscaling to a similar magnitude as exposure to TTX. Moreover, upscaling induced by DNMT inhibition required transcription; the RNA polymerase inhibitor actinomycin D blocked upscaling induced by DNMT inhibition. Knocking down the cytosine demethylase TET1 also blocked the upscaling effects of RG108. DNMT inhibition induced a multiplicative increase in mEPSC amplitude, indicating that the alterations in glutamate receptor abundance occurred in a coordinated manner throughout a neuron and were not limited to individual active synapses. Our data suggest that DNA methylation status controls transcription-dependent regulation of glutamatergic synaptic homeostasis. Furthermore, covalent DNA modifications may contribute to synaptic plasticity events that underlie the formation and stabilization of memories.
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Jarrod P
authors.initials: JP
authors.lastname: Meadows
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Mikael C
authors.initials: MC
authors.lastname: Guzman-Karlsson
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Phillips
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Cassie
authors.initials: C
authors.lastname: Holleman
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Posey
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Jeremy J
authors.initials: JJ
authors.lastname: Day
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA. jhablitz@uab.edu dsweatt@uab.edu.
authors.forename: John J
authors.initials: JJ
authors.lastname: Hablitz
authors.affiliation: Evelyn F. McKnight Brain Institute, Department of Neurobiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA. jhablitz@uab.edu dsweatt@uab.edu.
authors.forename: J David
authors.initials: JD
authors.lastname: Sweatt
mesh_terms: D000818:Animals; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects / Q000502:physiology*; D000090205:DNA Methyltransferase 3A; D055785:Gene Knockdown Techniques; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010797:Phthalimides / Q000494:pharmacology; D051381:Rats; D013569:Synapses / Q000235:genetics / Q000378:metabolism*; D055366:Synaptic Potentials / Q000187:drug effects / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology*; D014364:Tryptophan / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D010797:Phthalimides; C503639:RG108; D013779:Tetrodotoxin; D014364:Tryptophan; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; C000619382:Dnmt1 protein, rat