Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
The tiny volume of two-photon excitation (2PE) dictates where the photoswitchable drug Alloswitch is photoisomerized to its active trans-isoform, to only excite cells situated at the focal plane (0 µm).
Pittolo S, Lee H, Llado A, Tosi S, Bosch M, Bardia L, Gomez-Santacana X, Llebaria A, Soriano E, Colombelli J, Poskanzer KE, Perea G, Gorostiza P.
Proc Natl Acad Sci U S A 2019 Jul; 116: 13680.
The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and 2-photon excitation using pulsed near-infrared lasers to reversibly silence metabotropic glutamate 5 (mGlu5) receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 microm. Thus, 2-photon pharmacology using alloswitch allows investigating mGlu5-dependent processes in wild-type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles.
PubMed: 31196955. Doi: 10.1073/pnas.1900430116. Free PMC
