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Collection: Distribution of ICN Neurons in Male and Female 3D Reconstructed Rat Hearts

Study purpose: The purpose of this study was to establish a comprehensive 3D atlas of the rat intrinsic cardiac nervous system (ICN) at single-cell resolution to characterize the distribution patterns of intrinsic cardiac neurons between individuals and across sexes. To this end, we used a combination of high-throughput sectioning and imaging with a Knife Edge Scanning Microscope (KESM) to assemble image volumes and the Tissue Mapper software to map the location of ICN neurons and to annotate key cardiac anatomy for 3D heart reconstructions. Data collection: Hearts were fixed and then whole-mount diffusion stained with Cresyl Echt Violet to visualize ICN neurons. The hearts were then sectioned and imaged using a Knife Edge Scanning Microscope (KESM) to produce image stacks. Individual hearts were cut in one of three different orientations. Single neurons were mapped, and cardiac anatomy was segmented on select sections of the image stacks with the Tissue Mapper software. The spatial distribution of individually marked neurons and annotated heart anatomy were then visualized using a 3D reconstructed model of the heart. Primary conclusion: The distribution of neurons in male rat hearts showed similar patterns with three large clusters, despite the difference in sectioning orientations. The average number of mapped neurons was 2563 neurons in males and 1350 neurons in females. While the female hearts had fewer neurons, the distribution of neurons followed a similar pattern to the males along the interatrial sulcus. The reduced number may be due in part to experimental artifacts, but offers a preliminary insight of the ICN of males and females.
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