Sympathetic Neurons Regulate Cardiomyocyte Maturation in Culture.

William J Kowalski,Yongshun Lin,Wenling Li,Yoh-Suke Mukouyama,Chulan Kwon,Jizhong Zou,Hideki Uosaki,Emmanouil Tampakakis,Kira Patterson,Iris H Garcia-Pak

doi:10.3389/fcell.2022.850645

Abstract

Embryos devoid of autonomic innervation suffer sudden cardiac death. However, whether autonomic neurons have a role in heart development is poorly understood. To investigate if sympathetic neurons impact cardiomyocyte maturation, we co-cultured phenotypically immature cardiomyocytes derived from human induced pluripotent stem cells with mouse sympathetic ganglion neurons. We found that 1) multiple cardiac structure and ion channel genes related to cardiomyocyte maturation were up-regulated when co-cultured with sympathetic neurons; 2) sarcomere organization and connexin-43 gap junctions increased; 3) calcium imaging showed greater transient amplitudes. However, sarcomere spacing, relaxation time, and level of sarcoplasmic reticulum calcium did not show matured phenotypes. We further found that addition of endothelial and epicardial support cells did not enhance maturation to a greater extent beyond sympathetic neurons, while administration of isoproterenol alone was insufficient to induce changes in gene expression. These results demonstrate that sympathetic neurons have a significant and complex role in regulating cardiomyocyte development.

Highlights

The heart is densely innervated by sympathetic nerves, which extend from cell bodies housed in the sympathetic ganglia derived from trunk neural crest cells (Kimura et al, 2012)
We found that co-culture with sympathetic neurons (SNs) up-regulated multiple genes for CM maturation, ranging from myofiber components to ion channels and calcium machinery
To understand how SNs support developing CMs, we established a long-term culture system (30 days) of GFP-expressing hiPSCderived CMs (Lin et al, 2017) alone and with freshly isolated SNs dissected from E13.5 mouse embryos (Figure 1A)

Summary

Introduction

The heart is densely innervated by sympathetic nerves, which extend from cell bodies housed in the sympathetic ganglia derived from trunk neural crest cells (Kimura et al, 2012). In pediatric patients, sympathetic defects have been linked to sudden infant death syndrome, certain congenital heart defects, cardiac arrhythmic death, and hypertension (Kahn et al, 2003; Crump et al, 2011; Ohuchi et al, 2011). These innervation disorders may have foundations in the interactions between cardiomyocytes (CMs) and sympathetic neurons (SNs) during development. Understanding the function of the cardiac sympathetic nervous system in Sympathetic Regulation of Cardiomyocyte Maturation the embryo and its role in heart development would give valuable insight into fetal, pediatric, and adult heart disease

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Results

Conclusion