Accéder au contenu
Merck

RAI1 Regulates Activity-Dependent Nascent Transcription and Synaptic Scaling.

Cell reports (2020-08-14)
Patricia M Garay, Alex Chen, Takao Tsukahara, Jean Carlos Rodríguez Díaz, Rafi Kohen, J Christian Althaus, Margarete A Wallner, Roman J Giger, Kevin S Jones, Michael A Sutton, Shigeki Iwase
RÉSUMÉ

Long-lasting forms of synaptic plasticity such as synaptic scaling are critically dependent on transcription. Activity-dependent transcriptional dynamics in neurons, however, remain incompletely characterized because most previous efforts relied on measurement of steady-state mRNAs. Here, we use nascent RNA sequencing to profile transcriptional dynamics of primary neuron cultures undergoing network activity shifts. We find pervasive transcriptional changes, in which ∼45% of expressed genes respond to network activity shifts. We further link retinoic acid-induced 1 (RAI1), the Smith-Magenis syndrome gene, to the transcriptional program driven by reduced network activity. Remarkable agreement among nascent transcriptomes, dynamic chromatin occupancy of RAI1, and electrophysiological properties of Rai1-deficient neurons demonstrates the essential roles of RAI1 in suppressing synaptic upscaling in the naive network, while promoting upscaling triggered by activity silencing. These results highlight the utility of bona fide transcription profiling to discover mechanisms of activity-dependent chromatin remodeling that underlie normal and pathological synaptic plasticity.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Albumine de sérum bovin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Anticorps monoclonal anti-β-actine antibody produced in mouse, clone AC-15, ascites fluid
Sigma-Aldrich
Anticorps anti-NeuN, clone A60, clone A60, Chemicon®, from mouse
Sigma-Aldrich
Anticorps anti-Olig-2, Chemicon®, from rabbit
Sigma-Aldrich
Réactif TRI®, BD, For processing whole blood, plasma, or serum.
Sigma-Aldrich
Anticorps de chèvre anti-IgG de lapin, conjugué à de la peroxydase, 1 mg/mL (after reconstitution), Chemicon®
Sigma-Aldrich
Anticorps anti-PSD95 (protéine de densité post-synaptique 95), clone 6G6-1C9, clone 6G6-1C9, Chemicon®, from mouse
Sigma-Aldrich
Anticorps anti-MAP2, Chemicon®, from chicken
Sigma-Aldrich
5-Bromouridine, 98%
Sigma-Aldrich
MISSION® TRC2 pLKO.5-puro Non-Mammalian shRNA Control Plasmid DNA, Targets no known mammalian genes
Sigma-Aldrich
Anti-GluR1 Antibody, from rabbit, purified by affinity chromatography
Sigma-Aldrich
Rabbit Anti-Mouse IgG Antibody, F(ab′)2, HRP conjugate, Chemicon®, from rabbit