title: Anticonvulsant and anesthetic effects of a fluorescent neurosteroid analog activated by visible light.
abstract: Most photoactivatable compounds suffer from the limitations of the ultraviolet wavelengths that are required for activation. We synthesized a neuroactive steroid analog with a fluorescent (7-nitro-2,1,3-benzoxadiazol-4-yl) amino (NBD) group in the beta configuration at the C2 position of (3alpha,5alpha)-3-hydroxypregnan-20-one (allopregnanolone, 3alpha5alphaP). Light wavelengths (480 nm) that excite compound fluorescence strongly potentiate GABAA receptor function. Potentiation is limited by photodepletion of the receptor-active species. Photopotentiation is long-lived and stereoselective and shows single-channel hallmarks similar to steroid potentiation. Other NBD-conjugated compounds also generate photopotentiation, albeit with lower potency. Thus, photopotentiation does not require a known ligand for neurosteroid potentiating sites on the GABAA receptor. Photoactivation of a membrane-impermeant, fluorescent steroid analog demonstrates that membrane localization is critical for activity. The photoactivatable steroid silences pathological spiking in cultured rat hippocampal neurons and anesthetizes tadpoles. Fluorescent steroids photoactivated by visible light may be useful for modulating GABAA receptor function in a spatiotemporally defined manner.
authors.affiliation: Department of Neurology, Washington University School of Medicine, 660 S. Euclid Ave, St. Louis, Missouri 63110, USA.
authors.forename: Lawrence N
authors.initials: LN
authors.lastname: Eisenman
authors.forename: Hong-Jin
authors.initials: HJ
authors.lastname: Shu
authors.forename: Gustav
authors.initials: G
authors.lastname: Akk
authors.forename: Cunde
authors.initials: C
authors.lastname: Wang
authors.forename: Brad D
authors.initials: BD
authors.lastname: Manion
authors.forename: Geraldine J
authors.initials: GJ
authors.lastname: Kress
authors.forename: Alex S
authors.initials: AS
authors.lastname: Evers
authors.forename: Joe Henry
authors.initials: JH
authors.lastname: Steinbach
authors.forename: Douglas F
authors.initials: DF
authors.lastname: Covey
authors.forename: Charles F
authors.initials: CF
authors.lastname: Zorumski
authors.forename: Steven
authors.initials: S
authors.lastname: Mennerick
mesh_terms: D000777:Anesthetics / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D000927:Anticonvulsants / Q000494:pharmacology*; D002461:Cell Line, Transformed; D019793:Fluorescein; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D007814:Larva / Q000187:drug effects; D008027:Light; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D009474:Neurons / Q000187:drug effects; D018408:Patch-Clamp Techniques; D011280:Pregnanolone / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000494:pharmacology*; D021122:Protein Subunits / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000187:drug effects / Q000378:metabolism; D013329:Structure-Activity Relationship; D013550:Swimming; D014162:Transfection; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
chemical_list: D000777:Anesthetics; D000927:Anticonvulsants; D021122:Protein Subunits; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid; D011280:Pregnanolone; D019793:Fluorescein