Identification of a fluorescent general anesthetic, 1-aminoanthracene

Proc Natl Acad Sci U S A. 2009 Apr 21;106(16):6501-6. doi: 10.1073/pnas.0810590106. Epub 2009 Apr 3.

Abstract

We identified a fluorophore, 1-aminoanthracene (1-AMA), that is anesthetic, potentiates GABAergic transmission, and gives an appropriate dissociation constant, K(d) approximately 0.1 mM, for binding to the general anesthetic site in horse spleen apoferritin (HSAF). 1-AMA fluorescence is enhanced when bound to HSAF. Thus, displacement of 1-AMA from HSAF by other anesthetics attenuates the fluorescence signal and allows determination of K(d), as validated by isothermal titration calorimetry. This provides a unique fluorescence assay for compound screening and anesthetic discovery. Additional electrophysiology experiments in isolated cells indicate that 1-AMA potentiates chloride currents elicited by GABA, similar to many general anesthetics. Furthermore, 1-AMA reversibly immobilizes stage 45-50 Xenopus laevis tadpoles (EC(50) = 16 microM) and fluorescence micrographs show 1-AMA localized to brain and olfactory regions. Thus, 1-AMA provides an unprecedented opportunity for studying general anesthetic distribution in vivo at the cellular and subcellular levels.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anesthetics, General / metabolism*
  • Animals
  • Anthracenes / metabolism*
  • Anthracenes / pharmacology
  • Apoferritins / chemistry
  • Apoferritins / metabolism
  • Binding Sites
  • Chloride Channels / metabolism
  • Fluorescence
  • Fluorescent Dyes / metabolism*
  • Horses
  • Ion Channel Gating / drug effects
  • Isoflurane / metabolism
  • Larva / cytology
  • Larva / drug effects
  • Microscopy, Confocal
  • Neurons / cytology
  • Neurons / drug effects
  • Propofol / metabolism
  • Protein Structure, Secondary
  • Reflex, Startle / drug effects
  • Spectrometry, Fluorescence
  • Temperature
  • Xenopus laevis / metabolism
  • gamma-Aminobutyric Acid / pharmacology

Substances

  • Anesthetics, General
  • Anthracenes
  • Chloride Channels
  • Fluorescent Dyes
  • gamma-Aminobutyric Acid
  • 1-aminoanthracene
  • Apoferritins
  • Isoflurane
  • Propofol