3D single cell scale anatomical map of sex-dependent variability of the rat intrinsic cardiac nervous system

Abstract

We developed and analyzed a single cell scale anatomical map of the rat intrinsic cardiac nervous system (ICNS) across 4 male and 3 female hearts. We find the ICNS has a reliable structural organizational plan across individuals that may provide the foundation for further analyses of the ICNS in cardiac function and disease. The distribution of the ICNS was evaluated by 3D visualization and using data-driven clustering. The pattern, distribution and clustering of ICNS neurons across all male and female rat hearts is highly conserved, demonstrating a coherent organizational plan in which distinct clusters of neurons are consistently localized. Female hearts had fewer neurons, lower packing density and slightly reduced distribution, but with the same localization. We registered the anatomical data from each heart to a geometric scaffold, normalizing their 3D coordinates for the standardization of common anatomical planes and providing the path where multiple experimental results and data types can be integrated and compared.

The SPARC DRC: Building a Resource for the Autonomic Nervous System Community

“…With the scaffold in the fitted geometric configuration for a specimen, all other data such as digitized neurons and pathways in that configuration can be in material coordinates, or data can be used to fit continuous fields. Once this data registration is complete, data from multiple specimens can be compared in any common coordinate system over the scaffold, for example the idealized reference coordinates for the scaffold (Leung et al, 2020 ), or the average coordinates across multiple specimens (Osanlouy et al, 2020 ).…”

“…Cardiac regions and single ICN were segmented and annotated using the MBF Tissue Mapper software 25 . These data were used to generate and customize individual heart scaffolds to enable the registration of ICNs (Leung et al, 2020 ). Since ICNs are mainly distributed around the basal part of the heart and pulmonary veins, a “generic” scaffold was algorithmically generated from a set of anatomical and mathematical parameters for the atrial topology using the Scaffold-Maker software.…”

The SPARC DRC: Building a resource for the autonomic nervous system community

“…With the scaffold in the fitted geometric configuration for a specimen, all other data such as digitized neurons and pathways in that configuration can be in material coordinates, or data can be used to fit continuous fields. Once this data registration is complete, data from multiple specimens can be compared in any common coordinate system over the scaffold, for example the idealized reference coordinates for the scaffold (Leung et al, 2020), or the average coordinates across multiple specimens (Osanlouy et al, 2020).…”

“…Cardiac regions and single ICN were segmented and annotated using the MBF Tissue Mapper software 29 . These data were used to generate and customize individual heart scaffolds to enable the registration of ICNs (Leung et al, 2020). Since ICNs are mainly distributed around the basal part of the heart and pulmonary veins, a ‘generic’ scaffold was algorithmically generated from a set of anatomical and mathematical parameters for the atrial topology using the Scaffold-Maker software.…”

“…Here the heart is visualized from a superior angle to appreciate how the neuron locations in the original data are projected onto the generic scaffold. Figure adapted from (Leung et al, 2020).…”