Abstract
Analysis of calcium sparks in cardiomyocytes can provide valuable information about functional changes of calcium handling in health and disease. As a part of the calcium sparks analysis, sparks detection and characterization is necessary. Here, we describe a new open-source platform for automatic calcium sparks detection from line scan confocal images. The developed software is tailored for detecting only calcium sparks, allowing us to design a graphical user interface specifically for this task. The software enables detecting sparks automatically as well as adding, removing, or adjusting regions of interest marking each spark. The results of the analysis are stored in an SQL database, allowing simple integration with statistical tools. We have analyzed the performance of the algorithm using a large set of synthetic images with varying spark sizes and noise levels and also compared the analysis results with results obtained by software established in the field. The use of our software is illustrated by an analysis of the effect of isoprenaline (ISO) on spark frequency, amplitude, and spatial and temporal characteristics. For that, cardiomyocytes from C57BL/6 mice were used. We demonstrated an increase in spark frequency, tendency of having larger spark amplitudes, sparks with a longer duration, and occurrence of multiple sparks from the same site in the presence of ISO. We also show that the duration and the width of sparks with the same amplitude were similar in the absence and presence of ISO. The software was released as an open source repository and is available for free use and collaborative development.
Highlights
1901-Pos Non-ryanodine receptors (RyRs) Calcium Leak of the Sarcoplasmic Reticulum is Governed by TRPC1 in Cardiomyocytes Azmi A
Recording vacuoles enlarged by vacuolin-1 or PIKfyve inhibitors, YM201636 and APY-0201, from HEK293 cells expressing human Two Pore Channel 2 (TPC2), we investigated the interdependence between PI(3,5)P2 and nicotinic acid adenine dinucleotide phosphate (NAADP) on TPC2 activation
We further suggest that endogenous PI(3,5)P2 and NAADP contribute to the development of TPC2 currents in wholeendolysosomal recordings, which should be taken into account in data interpretation
Summary
1901-Pos Non-RyR Calcium Leak of the Sarcoplasmic Reticulum is Governed by TRPC1 in Cardiomyocytes Azmi A. Principal Component Analysis (PCA) indicated that the increase in [Ca2þ]cyto and frequency of local Ca2þevents are strongly associated with the HH condition while the altered responses to stimulation are not. Two Pore Channel 2 (TPC2) has been implicated in releasing Ca2þ from lysosomes in response to nicotinic acid adenine dinucleotide phosphate (NAADP). Recording vacuoles enlarged by vacuolin-1 or PIKfyve inhibitors, YM201636 and APY-0201, from HEK293 cells expressing human TPC2, we investigated the interdependence between PI(3,5)P2 and NAADP on TPC2 activation.
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