Abstract
Optogenetic technology has enabled unparalleled insights into cellular and organ physiology by providing exquisite temporal and spatial control of biological pathways. Here, an optogenetic approach is presented for selective activation of the intrinsic cardiac nervous system in excised perfused mouse hearts. The breeding of transgenic mice that have selective expression of channelrhodopsin in either catecholaminergic or cholinergic neurons is described. An approach for perfusing hearts excised from those animals, recording the ECG to measure heart rate changes, and an illumination technique using a custom micro-LED light source to activate channelrhodopsin is explained. We have used these methods in ongoing studies of the kinetics of autonomic control of cardiac electrophysiology and contractility, demonstrating the proven utility of optogenetic technology to enable unparalleled spatiotemporal anatomic-functional probing of the intrinsic cardiac nervous system.
Key words
- Autonomic nerves
- Cardiac function
- Electrophysiology
- Transgenic mice
- Optogenetics
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Acknowledgments
The microscopy expertise of Anastas Popratiloff, MD, PhD and the facilities GWU SEH Microscopy Core Facility are gratefully acknowledged. Matthew Stoyek, PhD is gratefully acknowledged for guidance in antibody staining of ChAT. Matthew Colonnese, PhD and Marnie Phillips, PhD are gratefully acknowledged for providing mice used in early experiments. We also thank Emilia Entcheva, PhD for many insightful discussions regarding photoactivation of channelrhodopsin within cardiac tissues.
Funding: This work was supported by grants from the National Institutes of Health (R21-HL132618, R01-HL133862, R01- HL141470), and a Don J. Levy and Elma Levy fellowship.
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Moreno, A., Kowalik, G., Mendelowitz, D., Kay, M.W. (2021). Optogenetic Control of Cardiac Autonomic Neurons in Transgenic Mice. In: Dempski, R. (eds) Channelrhodopsin. Methods in Molecular Biology, vol 2191. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0830-2_18
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DOI: https://doi.org/10.1007/978-1-0716-0830-2_18
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