Fixed samples of human atria and ventricles were sectioned and immunostained for neural markers using the ABC method. Specific targets were tyrosine hydroxylase, vesicular ACh transporter, and PGP9.5. Regional nerve densities were quantified using ImageJ.
Study Purpose: There are major gaps in our knowledge regarding innervation of the human heart by parasympathetic and sympathetic branches of the autonomic nervous system. Our goals for this study were to 1) generate a comprehensive and quantitative assessment of the innervation of atrial and ventricular regions of normal human heart and 2) determine if regional innervation is altered in patients with non-ischemic cardiomyopathy.
Data Collection: Digital images of stained sections were collected at 20X magnification, and nerve density in each field was quantified as percent area using ImageJ.
Primary Conclusion: Nerves were present and prominent throughout the human heart, with highest densities occurring in atrial regions, particularly the sinoatrial node. Cholinergic nerves were more abundant in atrial compared to ventricular regions. Sympathetic innervation density was more consistent throughout the heart, and noradrenergic nerves were 5-10 times more abundant than cholinergic nerves in most ventricular regions. This contrasts with the atria, where cholinergic nerves were more abundant than noradrenergic nerves at some locations. Within ventricular regions, the gap between cholinergic and noradrenergic nerve densities was less in right ventricular regions including the outflow tract. PGP5.5+ nerves where usually more abundant than TH+ and VAChT+ nerves. PGP9.5 labeling, sometimes intense, was also observed in some myocytes. Innervation was generally preserved in diseased hearts, but noradrenergic innervation appeared to be expanded in the ventricle on one heart, suggesting the possibility for compensatory remodeling.
Experimental Design: Perfusion or immersion fixed samples of human hearts were received, and frozen sections from each sample were cut and mounted on slides using a protocol that resulted in several representative sets of sections for each region. One set of sections for each region was immunostained for a specific neural marker using the brightfield ABC method of immunohistochemistry with ImpactVIP as the chromogen. Cholinergic and noradrenergic nerves were labeled using antibodies to the vesicular acetylcholine transporter (VAChT) and tyrosine hydroxylase (TH), respectively. Total innervation was evaluated using an antibody to protein gene product 9.5 (PGP9.5). Digital images of stained sections were collected at 20X magnification, and nerve density in each field was quantified as percent area using ImageJ. Extensive sampling was performed in two normal hearts and two diseased hearts (non-ischemic cardiomyopathy). This data is presented as tabulations of mean nerve density per sample. Specific right atrial regions, including the sinoatrial node, were evaluated in more hearts, and this data is presented as bar graphs with statistical comparison of VAChT and TH nerve densities.
Completeness: This dataset is complete.
Subjects & Samples: Heart samples from male (n=1) and female (n=3) human donors were used in this study.
Primary vs derivative data: Primary data folder contains digital images of the stained sections in the human heart. The images are organized in folders by the subject ID and sample name (region of the heart), respectively. These pictures are JPGs. Full scans (10x) of several tissue slides are also included. Nerve density information derived from these photos is located in the "derivative" folder. There is one Microsoft Excel spreadsheet for each donor and a sheet for each heart region within its corresponding donor's spreadsheet. Images in derivative folders are primary images that were converted with 20:1 compression to JPEG2000 (.jpx) by MBF Bioscience for web streaming and visualization on the SPARC Data Portal. Those images were also converted with lossless compression to OME-TIFF (.tif) by MBF Bioscience.
Important Notes: Information about the donors, tissue preparation, and tissue dissection are located in the "docs" folder, separated by donor and region. Pictures of the gross anatomy are uploaded as PDFs and include the relevant donor and preparation information. The nature of the preparation and dissection of the tissue from subjects Exp001 and Exp002 did not allow many gross anatomy photos.
0 - 0 of 0 files