A Diacylglycerol Photoswitching Protocol for Studying TRPC Channel Functions in Mammalian Cells and Tissue Slices

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2021 May 24

PMID 34027485

Citations 4

Authors

Trese Leinders-Zufall

Ursula Storch

Michael Mederos Y Schnitzler

Navin K Ojha

Kohei Koike

Thomas Gudermann

Frank Zufall

Affiliations

Soon will be listed here.

Abstract

Small molecular probes designed for photopharmacology and opto-chemogenetics are rapidly gaining widespread recognition for investigations of transient receptor potential canonical (TRPC) channels. This protocol describes the use of three photoswitchable diacylglycerol analogs-PhoDAG-1, PhoDAG-3, and OptoDArG-for ultrarapid activation and deactivation of native TRPC2 channels in mouse vomeronasal sensory neurons and olfactory type B cells, as well as heterologously expressed human TRPC6 channels. Photoconversion can be achieved in mammalian tissue slices and enables all-optical stimulation and shutoff of TRPC channels. For complete details on the use and execution of this protocol, please refer to Leinders-Zufall et al. (2018).

Citing Articles

Photoswitchable TRPC6 channel activators evoke distinct channel kinetics reflecting different gating behaviors.

Keck M, Hermann C, Lutzel K, Gudermann T, Konrad D, Mederos Y Schnitzler M iScience. 2024; 27(10):111008.

PMID: 39403196 PMC: 11472632. DOI: 10.1016/j.isci.2024.111008.

Photolipid excitation triggers depolarizing optocapacitive currents and action potentials.

Bassetto Jr C, Pfeffermann J, Yadav R, Strassgschwandtner S, Glasnov T, Bezanilla F Nat Commun. 2024; 15(1):1139.

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Optical Activation of Photoswitchable TRPC Ligands in the Mammalian Olfactory System Using Laser Scanning Confocal Microscopy.

Ojha N, Zufall F, Leinders-Zufall T Methods Mol Biol. 2023; 2710:31-47.

PMID: 37688722 DOI: 10.1007/978-1-0716-3425-7_3.

BTDAzo: A Photoswitchable TRPC5 Channel Activator.

Muller M, Niemeyer K, Urban N, Ojha N, Zufall F, Leinders-Zufall T Angew Chem Int Ed Engl. 2022; 61(36):e202201565.

PMID: 35713469 PMC: 9542918. DOI: 10.1002/anie.202201565.

References

Hofmann T, Obukhov A, Schaefer M, Harteneck C, Gudermann T, Schultz G . Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol. Nature. 1999; 397(6716):259-63. DOI: 10.1038/16711. View

Pnevmatikakis E . Analysis pipelines for calcium imaging data. Curr Opin Neurobiol. 2018; 55:15-21. DOI: 10.1016/j.conb.2018.11.004. View

Leinders-Zufall T, Ishii T, Mombaerts P, Zufall F, Boehm T . Structural requirements for the activation of vomeronasal sensory neurons by MHC peptides. Nat Neurosci. 2009; 12(12):1551-8. DOI: 10.1038/nn.2452. View

Curcic S, Tiapko O, Groschner K . Photopharmacology and opto-chemogenetics of TRPC channels-some therapeutic visions. Pharmacol Ther. 2019; 200:13-26. DOI: 10.1016/j.pharmthera.2019.04.003. View

Kiepas A, Voorand E, Mubaid F, Siegel P, Brown C . Optimizing live-cell fluorescence imaging conditions to minimize phototoxicity. J Cell Sci. 2020; 133(4). DOI: 10.1242/jcs.242834. View

Leinders-Zufall T, Brennan P, Widmayer P, S P, Maul-Pavicic A, Jager M . MHC class I peptides as chemosensory signals in the vomeronasal organ. Science. 2004; 306(5698):1033-7. DOI: 10.1126/science.1102818. View

Chamero P, Weiss J, Alonso M, Rodriguez-Prados M, Hisatsune C, Mikoshiba K . Type 3 inositol 1,4,5-trisphosphate receptor is dispensable for sensory activation of the mammalian vomeronasal organ. Sci Rep. 2017; 7(1):10260. PMC: 5579292. DOI: 10.1038/s41598-017-09638-8. View

Brechbuhl J, Luyet G, Moine F, Rodriguez I, Broillet M . Imaging pheromone sensing in a mouse vomeronasal acute tissue slice preparation. J Vis Exp. 2011; (58). PMC: 3369656. DOI: 10.3791/3311. View

Frank J, Yushchenko D, Hodson D, Lipstein N, Nagpal J, Rutter G . Photoswitchable diacylglycerols enable optical control of protein kinase C. Nat Chem Biol. 2016; 12(9):755-62. PMC: 6101201. DOI: 10.1038/nchembio.2141. View

10.

Fischer A, Jacobson K, Rose J, Zeller R . Preparation of slides and coverslips for microscopy. CSH Protoc. 2011; 2008:pdb.prot4988. DOI: 10.1101/pdb.prot4988. View

11.

Chen X, Sooch G, Demaree I, White F, Obukhov A . Transient Receptor Potential Canonical (TRPC) Channels: Then and Now. Cells. 2020; 9(9). PMC: 7565274. DOI: 10.3390/cells9091983. View

12.

Leinders-Zufall T, Storch U, Bleymehl K, Mederos Y Schnitzler M, Frank J, Konrad D . PhoDAGs Enable Optical Control of Diacylglycerol-Sensitive Transient Receptor Potential Channels. Cell Chem Biol. 2017; 25(2):215-223.e3. DOI: 10.1016/j.chembiol.2017.11.008. View

13.

Bleymehl K, Perez-Gomez A, Omura M, Moreno-Perez A, Macias D, Bai Z . A Sensor for Low Environmental Oxygen in the Mouse Main Olfactory Epithelium. Neuron. 2016; 92(6):1196-1203. PMC: 5196021. DOI: 10.1016/j.neuron.2016.11.001. View

14.

Gurtovenko A, Anwar J . Modulating the structure and properties of cell membranes: the molecular mechanism of action of dimethyl sulfoxide. J Phys Chem B. 2007; 111(35):10453-60. DOI: 10.1021/jp073113e. View

15.

Lichtenegger M, Tiapko O, Svobodova B, Stockner T, Glasnov T, Schreibmayer W . An optically controlled probe identifies lipid-gating fenestrations within the TRPC3 channel. Nat Chem Biol. 2018; 14(4):396-404. PMC: 5903546. DOI: 10.1038/s41589-018-0015-6. View

16.

Leinders-Zufall T, Lane A, Puche A, Ma W, Novotny M, Shipley M . Ultrasensitive pheromone detection by mammalian vomeronasal neurons. Nature. 2000; 405(6788):792-6. DOI: 10.1038/35015572. View

17.

Hull K, Morstein J, Trauner D . In Vivo Photopharmacology. Chem Rev. 2018; 118(21):10710-10747. DOI: 10.1021/acs.chemrev.8b00037. View

18.

Eckstein E, Pyrski M, Pinto S, Freichel M, Vennekens R, Zufall F . Cyclic regulation of Trpm4 expression in female vomeronasal neurons driven by ovarian sex hormones. Mol Cell Neurosci. 2020; 105:103495. DOI: 10.1016/j.mcn.2020.103495. View

19.

Notman R, Noro M, OMalley B, Anwar J . Molecular basis for dimethylsulfoxide (DMSO) action on lipid membranes. J Am Chem Soc. 2006; 128(43):13982-3. DOI: 10.1021/ja063363t. View

20.

Lucas P, Ukhanov K, Leinders-Zufall T, Zufall F . A diacylglycerol-gated cation channel in vomeronasal neuron dendrites is impaired in TRPC2 mutant mice: mechanism of pheromone transduction. Neuron. 2003; 40(3):551-61. DOI: 10.1016/s0896-6273(03)00675-5. View