Histone 4 acetylation and behavioral response at the Slo promoter region in Drosophila melanogaster after exposure to sevoflurane.

摘要

Today anesthetic gases remain the preferred pharmacologic agent for administration of general anesthesia. The exact mechanism of action of these agents is still unknown although several theories have been suggested. New fields of biology, including epigenetics and pharmacogenomics, provide promise for further describing the effects of these agents. Epigenetics, which literally translates to "above the genome," explores the relationship between environmental exposures and changes in gene expression. This study utilized four experiments to describe the epigenetic and behavioral effects of sevoflurane on Drosophila melanogaster, specifically at the Slopoke gene. The Slopoke gene is of interest as it has been previously described as active in the development of tolerance to ethanol, as well as other volatile agents. Two observational experiments were completed to investigate effects of the Slopoke gene on Drosophila specifically focusing on recovery time after sevoflurane anesthesia, time of day anesthetized, multiple exposures, and various recovery time periods between exposures. The epigenetic effects of sevoflurane exposure were quantified by Chromatin Immunoprecipitation followed by Real-Time Polymerase Chain Reaction. DNA histone 4 acetylation at the Ser-1/Lys-5/Lys-8/Lys-12 was quantified at both glyceraldehyde 3-phosphate dehydrogenase and Slopoke C0 promoter regions. This study found that after 30 minutes of exposure to 2%sevoflurane, and a 6 hour recovery period, the histone-four acetylation levels at both glyceraldehyde 3-phosphate dehydrogenase and the C0 promoter sites were significantly reduced. RNA expression for the Slopoke gene was also quantified by Reverse-Transcriptase Polymerase Chain Reaction. Slopoke RNA extraction corresponded with histone-four results, indicating that Slo RNA is reduced in the presence of sevoflurane. Our findings suggest further research on the epigenetic effects of sevoflurane at the Slopoke gene, as well as exploration of the implications that sevoflurane may act as a global deacetylator, are warranted.