title: Optically selective two-photon uncaging of glutamate at 900 nm.
abstract: We have synthesized a 7-diethylaminocoumarin (DEAC) derivative that allows wavelength-selective two-photon uncaging at 900 nm versus 720 nm. This new caging chromophore, called DEAC450, has an extended π-electron moiety at the 3-position that shifts the absorption spectrum maximum of DEAC from 375 to 450 nm. Two-photon excitation at 900 nm was more than 60-fold greater than at 720 nm. Two-photon uncaging of DEAC450-Glu at 900 nm at spine heads on pyramidal neurons in acutely isolated brain slices generated postsynaptic responses that were similar to spontaneous postsynaptic excitatory miniature currents, whereas significantly higher energies at 720 nm evoked no currents. Since many nitroaromatic caged compounds are two-photon active at 720 nm, optically selective uncaging of DEAC450-caged biomolecules at 900 nm may allow facile two-color optical interrogation of bimodal signaling pathways in living tissue with high resolution for the first time.
authors.affiliation: Department of Neuroscience, Mount Sinai School of Medicine, New York, New York 10029, USA.
authors.forename: Jeremy P
authors.initials: JP
authors.lastname: Olson
authors.forename: Hyung-Bae
authors.initials: HB
authors.lastname: Kwon
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Takasaki
authors.forename: Chiayu Q
authors.initials: CQ
authors.lastname: Chiu
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Higley
authors.forename: Bernardo L
authors.initials: BL
authors.lastname: Sabatini
authors.forename: Graham C R
authors.initials: GC
authors.lastname: Ellis-Davies
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D003374:Coumarins / Q000737:chemistry; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D018698:Glutamic Acid / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D051379:Mice; D059906:Neuroimaging / Q000379:methods*; D018408:Patch-Clamp Techniques; D010782:Photolysis; D017966:Pyramidal Cells / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013056:Spectrophotometry, Ultraviolet
chemical_list: D003374:Coumarins; D007202:Indicators and Reagents; D018698:Glutamic Acid