Direct modulation of microtubule stability contributes to anthracene general anesthesia

Daniel J. Emerson, Brian P. Weiser, John Psonis, Zhengzheng Liao, Olena Taratula, Ashley Fiamengo, Xiaozhao Wang, Keizo Sugasawa, Amos B. Smith, Roderic G. Eckenhoff, Ivan J. Dmochowski

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

Recently, we identified 1-aminoanthracene as a fluorescent general anesthetic. To investigate the mechanism of action, a photoactive analogue, 1-azidoanthracene, was synthesized. Administration of 1-azidoanthracene to albino stage 40-47 tadpoles was found to immobilize animals upon near-UV irradiation of the forebrain region. The immobilization was often reversible, but it was characterized by a longer duration consistent with covalent attachment of the ligand to functionally important targets. IEF/SDS-PAGE examination of irradiated tadpole brain homogenate revealed labeled protein, identified by mass spectrometry as β-tubulin. In vitro assays with aminoanthracene-cross-linked tubulin indicated inhibition of microtubule polymerization, similar to colchicine. Tandem mass spectrometry confirmed anthracene binding near the colchicine site. Stage 40-47 tadpoles were also incubated 1 h with microtubule stabilizing agents, epothilone D or discodermolide, followed by dosing with 1-aminoanthracene. The effective concentration of 1-aminoanthracene required to immobilize the tadpoles was significantly increased in the presence of either microtubule stabilizing agent. Epothilone D similarly mitigated the effects of a clinical neurosteroid general anesthetic, allopregnanolone, believed to occupy the colchicine site in tubulin. We conclude that neuronal microtubules are "on-pathway" targets for anthracene general anesthetics and may also represent functional targets for some neurosteroid general anesthetics.

Original languageEnglish
Pages (from-to)5389-5398
Number of pages10
JournalJournal of the American Chemical Society
Volume135
Issue number14
DOIs
StatePublished - 10 Apr 2013
Externally publishedYes

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