title: Sp3/REST/HDAC1/HDAC2 Complex Represses and Sp1/HIF-1/p300 Complex Activates ncx1 Gene Transcription, in Brain Ischemia and in Ischemic Brain Preconditioning, by Epigenetic Mechanism.
abstract: The Na(+)-Ca(2+) exchanger 1 (NCX1) is reduced in stroke by the RE1-silencing transcription factor (REST), whereas it is increased in ischemic brain preconditioning (PC) by hypoxia-inducible factor 1 (HIF-1). Because ncx1 brain promoter (ncx1-Br) has five putative consensus sequences, named Sp1A-E, for the specificity protein (Sp) family of transcription factors (Sp1-4), we investigated the role of this family in regulating ncx1 transcription in rat cortical neurons. Here we found that Sp1 is a transcriptional activator, whereas Sp3 is a transcriptional repressor of ncx1, and that both bind ncx1-Br in a sequence-specific manner, modulating ncx1 transcription through the Sp1 sites C-E. Furthermore, by transient middle cerebral artery occlusion (tMCAO) in rats, the transcriptional repressors Sp3 and REST colocalized with the two histone-deacetylases (HDACs) HDAC1 and HDAC2 on the ncx1-Br, with a consequent hypoacetylation. Contrarily, in PC+tMCAO the transcriptional activators Sp1 and HIF-1 colocalized with histone acetyltransferase p300 on ncx1-Br with a consequent hyperacetylation. In addition, in neurons silenced with siRNA of NCX1 and subjected to oxygen and glucose deprivation (OGD) (3 h) plus reoxygenation (RX) (24 h), the neuroprotection of Class I HDAC inhibitor MS-275 was counteracted, whereas in neurons overexpressing NCX1 and subjected to ischemic preconditioning (PC+OGD/RX), the neurotoxic effect of p300 inhibitor C646 was prevented. Collectively, these results demonstrate that NCX1 expression is regulated by the Sp3/REST/HDAC1/HDAC2 complex in tMCAO and by the Sp1/HIF-1/p300 complex in PC+tMCAO and that epigenetic intervention, by modulating the acetylation of ncx1-Br, may be a strategy for the development of innovative therapeutic intervention in stroke.
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy, Division of Pharmacology, Department of Science and Technology, University of Sannio, 82100 Benevento, Italy, and.
authors.forename: Luigi
authors.initials: L
authors.lastname: Formisano
authors.affiliation: IRCSS SDN, Naples 80131, Italy.
authors.forename: Natascia
authors.initials: N
authors.lastname: Guida
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Valsecchi
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Cantile
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Ornella
authors.initials: O
authors.lastname: Cuomo
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Vinciguerra
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Giusy
authors.initials: G
authors.lastname: Laudati
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pignataro
authors.affiliation: IRCSS SDN, Naples 80131, Italy.
authors.forename: Rossana
authors.initials: R
authors.lastname: Sirabella
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Di Renzo
authors.affiliation: Division of Pharmacology, Department of Neuroscience, Reproductive and Dentistry Sciences, School of Medicine, "Federico II" University of Naples, Naples 80131, Italy, IRCSS SDN, Naples 80131, Italy lannunzi@unina.it.
authors.forename: Lucio
authors.initials: L
authors.lastname: Annunziato
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D002545:Brain Ischemia / Q000473:pathology* / Q000517:prevention & control; D002540:Cerebral Cortex / Q000166:cytology; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044127:Epigenesis, Genetic; D056284:Histone Deacetylase 1 / Q000235:genetics; D006801:Humans; D019194:Ischemic Preconditioning; D008297:Male; D009474:Neurons / Q000378:metabolism*; D009578:Nitrobenzenes; D011720:Pyrazoles / Q000494:pharmacology; D047069:Pyrazolones; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins; D019831:Sodium-Calcium Exchanger / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection; D050880:p300-CBP Transcription Factors / Q000378:metabolism; D000099229:RE1-Silencing Transcription Factor
chemical_list: D001565:Benzoates; D004791:Enzyme Inhibitors; D056284:Histone Deacetylase 1; D009578:Nitrobenzenes; D011720:Pyrazoles; D047069:Pyrazolones; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D019831:Sodium-Calcium Exchanger; D050880:p300-CBP Transcription Factors; D000099229:RE1-Silencing Transcription Factor; C584509:4-(4-((5-(4,5-dimethyl-2-nitrophenyl)-2-furanyl)methylene)-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-1-yl)benzoic acid; C110374:sodium-calcium exchanger 1; C110550:HDAC1 protein, human
keywords: REST\NRSF; acetylation; brain ischemia; epigenetic; ischemic brain preconditioning; sodium/calcium exchanger