Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2019 Jul 2;116(27):13680-13689. doi: 10.1073/pnas.1900430116. Epub 2019 Jun 13.

Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology.

Author information

1
Institute for Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
2
Institute for Research in Biomedicine (IRB Barcelona), Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
3
Institute of Advanced Chemistry of Catalonia, Consejo Superior de Investigaciones Científicas (IQAC-CSIC), 08034 Barcelona, Spain.
4
Department of Cell Biology, Physiology, and Immunology, University of Barcelona (UB), 08028 Barcelona, Spain.
5
Catalan Institution for Research and Advanced Studies (ICREA), 08010 Barcelona, Spain.
6
Network Center of Biomedical Research in Neurodegenerative Diseases (CIBERNED), 28031 Madrid, Spain.
7
Department of Biochemistry & Biophysics, University of California, San Francisco (UCSF), CA 94158.
8
Kavli Institute for Fundamental Neuroscience, University of California, San Francisco, CA 94158.
9
Cajal Institute, Consejo Superior de Investigaciones Científicas (IC-CSIC), 28002 Madrid, Spain.
10
Institute for Bioengineering of Catalonia (IBEC), Barcelona Institute of Science and Technology, 08028 Barcelona, Spain; pau@icrea.cat.
11
Network Center of Biomedical Research in Bioengineering, Biomaterials, and Nanomedicine (CIBER-BBN), 50015 Zaragoza, Spain.

Abstract

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 µm. 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.

KEYWORDS:

2-photon pharmacology; functional silencing; pharmacological selectivity; photoactivation; photopharmacology

PMID:
31196955
PMCID:
PMC6613107
[Available on 2019-12-13]
DOI:
10.1073/pnas.1900430116

Supplemental Content

Full text links

Icon for HighWire
Loading ...
Support Center