Cardiomyocyte Proliferation Upon Heart Injury in Zebrafish

How do you visualize proliferating cardiomyocytes after heart injury and analyze cell responses during the recovery process? - MicaCam Episode 04 - Video On Demand

Zebrafish heart showing the ventricle with an injury in the lower area Zebrafish_heart_showing_ventricle_with_injury_teaser.jpg

This edition of MicaCam focuses on the study of the zebrafish (Danio rerio) whose heart cells, unlike the heart cells of other mammals, can fully regenerate after injury. Our special guest Laura Peces-Barba Castaño from the Max Planck Institute for Heart and Lung Research will analyse different parameters of this regeneration, such as cell proliferation. She will use live cell imaging data to look at differentially labelled cardiomyocytes and proliferating cells, following their response during recovery.

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Key Learnings

  • How to overcome the challenges of staining and imaging injured cardiac tissue
  • How to visualize and differentiate injured and proliferating cardiomyocytes
  • How to optimize the imaging and reconstruction of organ explants

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Speakers

Laura Peces-Barba Castaño    

PhD student - Prof. Dr. Stainier Department, Max Planck Institute for Heart and Lung Research

Laura holds a degree in Biomedical Sciences at the University of Aberdeen (Scotland). During this time, she also did an Industrial Placement at the Francis Crick Institute working with malaria parasites. Currently, Laura is undertaking a PhD in heart regeneration in the Stainier Department (Max Planck Institute), where she aims to discover the mechanisms leading to heart regeneration in the zebrafish. 

Dr. Lynne Turnbull, Senior Application Manager – Leica Microsystems

Lynne is a Senior Application Manager at Leica Microsystems. She received her PhD in Sydney Australia in cardiac biophysics and undertook postdoctoral training in San Francisco and Melbourne. Upon moving to the University of Technology Sydney, Lynne established and managed the Microbial Imaging Facility. Lynne joined Leica Microsystems in 2021 as a Senior Application Manager and is based at the EMBL IC in Heidelberg. 

The Experiment

MicaCam Episode 04 - original broadcast date: 25th May 2022

In this episode we talked with our guest Laura Peces-Barba Castaño about how hearts regenerate after injury in the model organism zebrafish. Mammalian hearts have a very limited capacity to regenerate after injury, leading to the presence of scar and fibrotic tissue. However, zebrafish hearts can regenerate after severe injury without scar formation, making the zebrafish a great experimental tool to explore cardiac injury.

Laura showed us the experimental procedure to injure the tip of a zebrafish heart and how she processed the hearts for imaging. We learned to identify the border zone between the injured and non-injured tissue and how this zone is the area of most interest for studying heart recovery. She used specific staining of cardiomyocytes, proliferating cells, and all cells to create differential staining patterns within the border zone. The differential staining patterns allowed the quantification of injured and proliferating cardiomyocytes. Our guest also described some of the challenges of processing, staining, and imaging the cardiac tissue sections.    

The ability of Mica’s sample finder to create a quick overview of the whole slide that contains 10 heart sections was a favorite feature for Laura, as it allowed easy navigation and exploration of the sections before deciding for which sections images were to be acquired. Tiled images of a single z-slice of each section could be captured rapidly in widefield and quickly examined. Then we decided about which sections are suitable for further observation with confocal imaging with or without z-stacks. This increased the efficiency of the workflow and time spent imaging. Laura was also very happy that we could do all this in a brightly lit room, instead of needing to be in a dark cave while doing microscopy!

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