A Matched-filter Based Algorithm for Subcellular Classification of T-system in Cardiac Tissues

Abstract

In mammalian ventricular cardiomyocytes, invaginations of the surface membrane form the transverse tubular system (T-system) which consists of transverse tubules (TTs) that align with sarcomeres and Z-lines as well as longitudinal tubules (LTs) that are present between Z-lines in some species. In many cardiac disease etiologies the T-system is perturbed, which is believed to promote spatially heterogeneous, dyssynchronous Ca2+ release and inefficient contraction. In general, T-system characterization approaches have been directed primarily at isolated cells and do not detect subcellular T-system heterogeneity. Here we present MatchedMyo, a matched-filter based algorithm for subcellular T-system characterization in isolated cardiomyocytes and millimeter-scale myocardial sections. The algorithm utilizes "filters" representative of TTs, LTs, and T-system absence. Application of the algorithm to cardiomyocytes isolated from rat disease models of myocardial infarction (MI), dilated cardiomyopathy induced via aortic banding (AB), and sham surgery confirmed and quantified heterogeneous T-system structure and remodeling. Cardiomyocytes from post-MI hearts exhibited increasing T-system disarray as proximity to the infarct increased. We found significant (p<0.05, Welchs t-test) increases in LT density within cardiomyocytes proximal to the infarct (12{+/-}3%, data reported as mean {+/-} SD, n=3) vs. sham (4{+/-}2%, n=5), but not distal to the infarct (7{+/-}1%, n=3). The algorithm also detected decreases in TTs within 5{degrees} of the myocyte minor axis for isolated AB (36{+/-}9%, n=3) and MI cardiomyocytes located intermediate (37{+/-}4%, n=3) and proximal (34{+/-}4%, n=3) to the infarct vs. sham (57{+/-}12%, n=5). Application of bootstrapping to rabbit MI tissue revealed distal sections comprised 18.9{+/-}1.0% TTs while proximal sections comprised 10.1{+/-}0.8% TTs (p<0.05), a 46.6% decrease. The matched filter approach therefore provides a robust and scalable technique for T-system characterization from isolated cells through millimeter-scale myocardial sections.