Drug-induced epigenetic changes produce drug tolerance.
Tolerance to drugs that affect neural activity is mediated, in part, by adaptive mechanisms that attempt to restore normal neural excitability. Changes in the expression of ion channel genes are thought to play an important role in these neural adaptations. The slo gene encodes the pore-forming subunit of BK-type Ca(2+)-activated K(+) channels, which regulate many aspects of neural activity. Given that induction of slo gene expression plays an important role in the acquisition of tolerance to sedating drugs, we investigated the molecular mechanism of gene induction. Using chromatin immunoprecipitation followed by real-time PCR, we show that a single brief sedation with the anesthetic benzyl alcohol generates a spatiotemporal pattern of histone H4 acetylation across the slo promoter region. Inducing histone acetylation with a histone deacetylase inhibitor yields a similar pattern of changes in histone acetylation, up-regulates slo expression, and phenocopies tolerance in a slo-dependent manner. The cAMP response element binding protein (CREB) is an important transcription factor mediating experience-based neuroadaptations. The slo promoter region contains putative binding sites for the CREB transcription factor. Chromatin immunoprecipitation assays show that benzyl alcohol sedation enhances CREB binding within the slo promoter region. Furthermore, activation of a CREB dominant-negative transgene blocks benzyl alcohol-induced changes in histone acetylation within the slo promoter region, slo induction, and behavioral tolerance caused by benzyl alcohol sedation. These findings provide unique evidence that links molecular epigenetic histone modifications and transcriptional induction of an ion channel gene with a single behavioral event.
Wang
Yan
Y
Section of Neurobiology, The University of Texas at Austin, Austin, Texas, USA.
Krishnan
Harish R
HR
Ghezzi
Alfredo
A
Yin
Jerry C P
JC
Atkinson
Nigel S
NS
D000107:Acetylation; D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D019905:Benzyl Alcohol / Q000494:pharmacology*; D002087:Butyrates / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster; D004361:Drug Tolerance / Q000502:physiology*; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects*; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D016415:Sequence Alignment; D012997:Solvents / Q000494:pharmacology; D015533:Transcriptional Activation; D019076:Transgenes
D000779:Anesthetics, Local; D002087:Butyrates; D017362:Cyclic AMP Response Element-Binding Protein; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D015221:Potassium Channels; D012997:Solvents; D006655:Histone Deacetylases; D019905:Benzyl Alcohol