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Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp

Sarcomeres are the basic contractile units within cardiac myocytes, and the collective shortening of sarcomeres aligned along myofibrils generates the force driving the heartbeat. The alignment of the individual sarcomeres is important for proper force generation, and misaligned sarcomeres are assoc...

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Autores principales: Neininger-Castro, Abigail C., Hayes, James B., Sanchez, Zachary C., Taneja, Nilay, Fenix, Aidan M., Moparthi, Satish, Vassilopoulos, Stéphane, Burnette, Dylan T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882215/
https://www.ncbi.nlm.nih.gov/pubmed/36711995
http://dx.doi.org/10.1101/2023.01.11.523681
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author Neininger-Castro, Abigail C.
Hayes, James B.
Sanchez, Zachary C.
Taneja, Nilay
Fenix, Aidan M.
Moparthi, Satish
Vassilopoulos, Stéphane
Burnette, Dylan T.
author_facet Neininger-Castro, Abigail C.
Hayes, James B.
Sanchez, Zachary C.
Taneja, Nilay
Fenix, Aidan M.
Moparthi, Satish
Vassilopoulos, Stéphane
Burnette, Dylan T.
author_sort Neininger-Castro, Abigail C.
collection PubMed
description Sarcomeres are the basic contractile units within cardiac myocytes, and the collective shortening of sarcomeres aligned along myofibrils generates the force driving the heartbeat. The alignment of the individual sarcomeres is important for proper force generation, and misaligned sarcomeres are associated with diseases including cardiomyopathies and COVID-19. The actin bundling protein, α-actinin-2, localizes to the “Z-Bodies” of sarcomere precursors and the “Z-Lines” of sarcomeres, and has been used previously to assess sarcomere assembly and maintenance. Previous measurements of α-actinin-2 organization have been largely accomplished manually, which is time-consuming and has hampered research progress. Here, we introduce sarcApp, an image analysis tool that quantifies several components of the cardiac sarcomere and their alignment in muscle cells and tissue. We first developed sarcApp to utilize deep learning-based segmentation and real space quantification to measure α-actinin-2 structures and determine the organization of both precursors and sarcomeres/myofibrils. We then expanded sarcApp to analyze “M-Lines” using the localization of myomesin and a protein that connects the Z-Lines to the M-Line (titin). sarcApp produces 33 distinct measurements per cell and 24 per myofibril that allow for precise quantification of changes in sarcomeres, myofibrils, and their precursors. We validated this system with perturbations to sarcomere assembly. We found perturbations that affected Z-Lines and M-Lines differently, suggesting that they may be regulated independently during sarcomere assembly.
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spelling pubmed-98822152023-01-28 Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp Neininger-Castro, Abigail C. Hayes, James B. Sanchez, Zachary C. Taneja, Nilay Fenix, Aidan M. Moparthi, Satish Vassilopoulos, Stéphane Burnette, Dylan T. bioRxiv Article Sarcomeres are the basic contractile units within cardiac myocytes, and the collective shortening of sarcomeres aligned along myofibrils generates the force driving the heartbeat. The alignment of the individual sarcomeres is important for proper force generation, and misaligned sarcomeres are associated with diseases including cardiomyopathies and COVID-19. The actin bundling protein, α-actinin-2, localizes to the “Z-Bodies” of sarcomere precursors and the “Z-Lines” of sarcomeres, and has been used previously to assess sarcomere assembly and maintenance. Previous measurements of α-actinin-2 organization have been largely accomplished manually, which is time-consuming and has hampered research progress. Here, we introduce sarcApp, an image analysis tool that quantifies several components of the cardiac sarcomere and their alignment in muscle cells and tissue. We first developed sarcApp to utilize deep learning-based segmentation and real space quantification to measure α-actinin-2 structures and determine the organization of both precursors and sarcomeres/myofibrils. We then expanded sarcApp to analyze “M-Lines” using the localization of myomesin and a protein that connects the Z-Lines to the M-Line (titin). sarcApp produces 33 distinct measurements per cell and 24 per myofibril that allow for precise quantification of changes in sarcomeres, myofibrils, and their precursors. We validated this system with perturbations to sarcomere assembly. We found perturbations that affected Z-Lines and M-Lines differently, suggesting that they may be regulated independently during sarcomere assembly. Cold Spring Harbor Laboratory 2023-10-06 /pmc/articles/PMC9882215/ /pubmed/36711995 http://dx.doi.org/10.1101/2023.01.11.523681 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Neininger-Castro, Abigail C.
Hayes, James B.
Sanchez, Zachary C.
Taneja, Nilay
Fenix, Aidan M.
Moparthi, Satish
Vassilopoulos, Stéphane
Burnette, Dylan T.
Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title_full Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title_fullStr Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title_full_unstemmed Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title_short Independent regulation of Z-lines and M-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcApp
title_sort independent regulation of z-lines and m-lines during sarcomere assembly in cardiac myocytes revealed by the automatic image analysis software sarcapp
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882215/
https://www.ncbi.nlm.nih.gov/pubmed/36711995
http://dx.doi.org/10.1101/2023.01.11.523681
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