Novel Computational Method for Measuring Cardiomyocyte Dimensions: Applied to Hypertrophic Calmodulin 1 Knock Out Mouse Model
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Date
2023-05
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The Ohio State University
Abstract
Cardiac hypertrophy is a remodeling response of the heart characterized by increased cardiomyocyte size and ventricular wall thickening. Calcium regulates cardiomyocyte functions and is bound by calmodulin which relays calcium signaling. Altered calcium signaling is part of the pathophysiology of cardiac hypertrophy, so we hypothesize that alterations in calmodulin activity may contribute to different cell morphologies in cardiac hypertrophy. To test this hypothesis, cardiomyocyte dimensions will be obtained; however, current techniques require manual measurements of individual cells which is time-consuming, creates subjectivity, and has limited throughput. A novel computational method was developed in MATLAB that automatically measures cardiomyocytes from images with pre-defined cell selection criteria. The code was developed using confocal microscopy images of adult mouse cardiomyocytes cultured in control and 20 μM phenylephrine conditions, measured cell length, width, and area, and was validated against manual measurements from ImageJ. The program segmented the images while reducing noise and excluded, measured, and globally compared cells from multiple images. There was no statistically significant difference in cardiomyocyte length and area between the code and ImageJ for either condition while there was for cell width. Hypertrophy was induced in WT and calmodulin 1 knock-out mice via transverse aortic constriction surgery and images were blindly run through the program. There was no statistically significant difference in length and area measurements between the groups so knocking out calmodulin 1 may not have an effect on cell morphology in cardiac hypertrophy. The novel computational program provides multiple advantages including automation, reproducibility, robust analysis of large data sets, and it has the potential to facilitate large-scale studies of hypertrophy. Overall, this study will allow the impacts of calmodulin in cardiac hypertrophy to be better understood.
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Keywords
Cardiac Hypertrophy, Cardiomyocytes, Measurements, Computational Method