Kinetic
analysis of dopamine receptor activation and inactivation
and the study of dopamine-dependent signaling requires precise simulation
of the presynaptic release of the neurotransmitter dopamine and tight
temporal control over the release of dopamine receptor antagonists.
The 8-cyano-7-hydroxyquinolinyl (CyHQ) photoremovable protecting group
was conjugated to dopamine and the dopamine receptor antagonist sulpiride
to generate “caged” versions of these neuromodulators
(CyHQ-O-DA and CyHQ-sulpiride, respectively) that
could release their payloads with 365 or 405 nm light or through 2-photon
excitation (2PE) at 740 nm. These compounds are stable under physiological
conditions in the dark, yet photolyze rapidly and cleanly to yield
dopamine or sulpiride and the caging remnant CyHQ–OH. CyHQ-O-DA mediated the light activation of dopamine-1 (D1) receptors
on the breast cancer cell line MDA-MB-231 in culture. In mouse brain
slice from the substantia nigra pars compacta, localized flash photolysis
of CyHQ-O-DA accurately mimicked the natural presynaptic
release of dopamine and activation of dopamine-2 (D2) receptors, causing
a robust, concentration-dependent, and repeatable G protein-coupled
inwardly rectifying potassium channel-mediated outward current in
whole-cell voltage clamp recordings that was amplified by cocaine
and blocked by sulpiride. Photolysis of CyHQ-sulpiride rapidly blocked
synaptic activity, enabling measurement of the unbinding rates of
dopamine and quinpirole, a D2 receptor agonist. These tools will enable
more detailed study of dopamine receptors, their interactions with
other GPCRs, and the physiology of dopamine signaling in the brain.