Fluorescent Anesthetics

Abstract

Methods for using exogenous fluorophore and general anesthetic 1-aminoanthracene (1-AMA) and its photoactive derivative 1-azidoanthracene (1-AZA) are provided. 1-AMA potentiates GABA_A chloride currents and immobilizes Xenopus laevis tadpoles. Cellular and tissue anesthetic distribution can be imaged for quantifying "on-pathway" and "off-pathway" targets. 1-AZA shares targets with 1-AMA and offers further optoanesthetic spatial and temporal control upon near-UV laser irradiation. Furthermore, 1-AZA adduction provides screening of possible relevant anesthetic protein targets and binding site characterization. We highlight several useful imaging and binding assays to demonstrate utility of 1-AMA and its derivative 1-AZA.

Keywords: 1-Aminoanthracene; 1-Azidoanthracene; Caged anesthetic; Confocal microscopy; Fluorescent probe.

Figure 1:

Fluorescence micrograph of 1-AMA distribution (15 (μM) in Xenopus laevis tadpole after incubation for 30 min in pond water. Reprinted with permission from Emerson, D.J., Liao, Z., Eckenhoff, R.G., Dmochowski, I.J. A Novel Fluorescent General Anesthetic Enables Imaging of Sites of Action In Vivo. Anesthesiology. 6 2012, Vol.l 16, 1363.

Figure 2.

Photoactivation of 1-AZA in vivo. Representative confocal fluorescence images of 1-AZA in a tadpole brain: before (A), during (B) and after (C) near-UV laser photolysis of a tadpole forebrain. The forebrain area was irradiated for 10 s by 351 and 364 nm lasers (100% power). Bottom plot shows the average fluorescent pixel intensity over time under 488 nm laser excitation. Reprinted with permission from Emerson, D J., Weiser, B.P., Psonis, J., Liao, Z.,Taratula, O., Fiamengo, A., Wang, X., Sugasawa, K., Smith, A.B., Eckenhoff, R.G., Dmochowski, I J. Direct Modulation of Microtubule Stability Contributes to Anthracene General Anesthesia. Journal of the American Chemical Society. Copy write 2013. American Chemical Society.