Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart

Wen, Q., Gandhi, K., Capel, Rebecca A., Hao, G., O'Shea, C., Neagu, G., Pearcey, S., Pavlovic, D., Terrar, Derek A., Wu, J. et al, Faggian, G., Camelliti, Patrizia and Lei, M. (2018) Transverse cardiac slicing and optical imaging for analysis of transmural gradients in membrane potential and Ca²⁺ transients in murine heart The Journal of Physiology, 596 (17). pp. 3951-3965.

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Abstract

Transmural and regional gradients in membrane potential and Ca²⁺ transient in the murine heart are largely unexplored. Here, we developed and validated a robust approach which combines transverse ultra‐thin cardiac slices and high resolution optical mapping to enable systematic analysis of transmural and regional gradients in transmembrane potential (Vm) and intracellular Ca²⁺ transient (CaT) across the entire murine ventricles. The voltage dye RH237 or Ca²⁺ dye Rhod‐2 AM were loaded through the coronary circulation using a Langendorff perfusion system. Short‐axis slices (300 μm thick) were prepared from the entire ventricles (from the apex to the base) by using a high‐precision vibratome. Action potentials (APs) and CaTs were recorded with optical mapping during steady‐state baseline and rapid pacing. Significant transmural gradients in V_m and CaT were observed in the left ventricle, with longer AP duration (APD₅₀ and APD₇₅) and CaT duration (CaTD₅₀ and CaTD₇₅) in the endocardium compared with that in the epicardium. No significant regional gradients were observed along the apico‐basal axis of the left ventricle. Interventricular gradients were detected with significantly shorter APD₅₀, APD₇₅ and CaTD₅₀ in the right ventricle compared with left ventricle and ventricular septum. During rapid pacing, AP and CaT alternans were observed in most ventricular regions, with significantly greater incidence in the endocardium in comparison with epicardium. In conclusion, these observations demonstrate the feasibility of our new approach to cardiac slicing for systematic analysis of intrinsic transmural and regional gradients in V_m and CaT in murine ventricular tissue.

Item Type:	Article
Divisions :	Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :	Name Email ORCID Wen, Q. Gandhi, K. Capel, Rebecca A. Hao, G. O'Shea, C. Neagu, G. Pearcey, S. Pavlovic, D. Terrar, Derek A. Wu, J. Faggian, G. Camelliti, Patrizia p.camelliti@surrey.ac.uk Lei, M.
Date :	21 June 2018
DOI :	10.1113/JP276239
Copyright Disclaimer :	© 2018 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Uncontrolled Keywords :	cardiac slices; optical imaging; murine heart; electrophysiological heterogeneity
Depositing User :	Clive Harris
Date Deposited :	30 Jul 2018 09:50
Last Modified :	08 Nov 2018 14:29
URI:	http://epubs.surrey.ac.uk/id/eprint/848801

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