Extended culture and imaging of normal and regenerating adult zebrafish hearts in a fluidic device

Zebrafish are excellent at regenerating their heart by reinitiating proliferation in pre-existing cardiomyocytes. Studying how zebrafish achieve this holds great potential in developing new strategies to boost mammalian heart regeneration. Nevertheless, the lack of appropriate live imaging tools for the adult zebrafish heart has limited detailed studies into the dynamics underlying cardiomyocyte proliferation. Here, we address this by developing a system in which cardiac slices of the injured zebrafish heart are cultured ex vivo for several days while retaining key regenerative characteristics including cardiomyocyte proliferation. In addition, we show that the cardiac slice culture system is compatible with live timelapse imaging and allows manipulation of regenerating cardiomyocytes with drugs that normally would have toxic effects that prevent its use. Finally, we use the cardiac slices to demonstrate that adult cardiomyocytes with fully assembled sarcomeres can partially disassemble their sarcomeres in a calpain and proteasome dependent manner to progress through nuclear division and cytokinesis. In conclusion, we have developed a cardiac slice culture system, which allows imaging of native cardiomyocyte dynamics in real time to discover cellular mechanisms during heart regeneration.

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Inhibition of let-7c Regulates Cardiac Regeneration after Cryoinjury in Adult Zebrafish

Journal of Cardiovascular Development and Disease ◽

10.3390/jcdd6020016 ◽

2019 ◽

Vol 6 (2) ◽

pp. 16 ◽

Cited By ~ 1

Author(s):

Suneeta Narumanchi ◽

Karri Kalervo ◽

Sanni Perttunen ◽

Hong Wang ◽

Katariina Immonen ◽

...

Keyword(s):

Cardiac Function ◽

Cardiac Regeneration ◽

Nuclear Antigen ◽

Heart Regeneration ◽

Adult Zebrafish ◽

Tissue Samples ◽

Micro Rnas ◽

Proliferating Cell ◽

Zebrafish Heart

The let-7c family of micro-RNAs (miRNAs) is expressed during embryonic development and plays an important role in cell differentiation. We have investigated the role of let-7c in heart regeneration after injury in adult zebrafish. let-7c antagomir or scramble injections were given at one day after cryoinjury (1 dpi). Tissue samples were collected at 7 dpi, 14 dpi and 28 dpi and cardiac function was assessed before cryoinjury, 1 dpi, 7 dpi, 14 dpi and 28 dpi. Inhibition of let-7c increased the rate of fibrinolysis, increased the number of proliferating cell nuclear antigen (PCNA) positive cardiomyocytes at 7 dpi and increased the expression of the epicardial marker raldh2 at 7 dpi. Additionally, cardiac function measured with echocardiography recovered slightly more rapidly after inhibition of let-7c. These results reveal a beneficial role of let-7c inhibition in adult zebrafish heart regeneration.

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High-frequency ultrasound deformation imaging for adult zebrafish during heart regeneration

Quantitative Imaging in Medicine and Surgery ◽

10.21037/qims.2019.09.20 ◽

2020 ◽

Vol 10 (1) ◽

pp. 66-75

Author(s):

Chen Ho-Chiang ◽

Hsin Huang ◽

Chih-Chung Huang

Keyword(s):

High Frequency ◽

Heart Regeneration ◽

Adult Zebrafish ◽

High Frequency Ultrasound ◽

Deformation Imaging ◽

Frequency Ultrasound

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High-frequency dual mode pulsed wave Doppler imaging for monitoring the functional regeneration of adult zebrafish hearts

Journal of The Royal Society Interface ◽

10.1098/rsif.2014.1154 ◽

2015 ◽

Vol 12 (103) ◽

pp. 20141154 ◽

Cited By ~ 8

Author(s):

Bong Jin Kang ◽

Jinhyoung Park ◽

Jieun Kim ◽

Hyung Ham Kim ◽

Changyang Lee ◽

...

Keyword(s):

Diastolic Dysfunction ◽

High Frequency ◽

Doppler Imaging ◽

Flow Mode ◽

Heart Regeneration ◽

Adult Zebrafish ◽

Dual Mode ◽

Doppler Flow ◽

Pulsed Wave Doppler ◽

Zebrafish Heart

Adult zebrafish is a well-known small animal model for studying heart regeneration. Although the regeneration of scars made by resecting the ventricular apex has been visualized with histological methods, there is no adequate imaging tool for tracking the functional recovery of the damaged heart. For this reason, high-frequency Doppler echocardiography using dual mode pulsed wave Doppler, which provides both tissue Doppler (TD) and Doppler flow in a same cardiac cycle, is developed with a 30 MHz high-frequency array ultrasound imaging system. Phantom studies show that the Doppler flow mode of the dual mode is capable of measuring the flow velocity from 0.1 to 15 cm s −1 with high accuracy ( p -value = 0.974 > 0.05). In the in vivo study of zebrafish, both TD and Doppler flow signals were simultaneously obtained from the zebrafish heart for the first time, and the synchronized valve motions with the blood flow signals were identified. In the longitudinal study on the zebrafish heart regeneration, the parameters for diagnosing the diastolic dysfunction, for example, E / E m < 10, E / A < 0.14 for wild-type zebrafish, were measured, and the type of diastolic dysfunction caused by the amputation was found to be similar to the restrictive filling. The diastolic function was fully recovered within four weeks post-amputation.

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Nrg1 is an injury-induced cardiomyocyte mitogen for the endogenous heart regeneration program in zebrafish

eLife ◽

10.7554/elife.05871 ◽

2015 ◽

Vol 4 ◽

Cited By ~ 111

Author(s):

Matthew Gemberling ◽

Ravi Karra ◽

Amy L Dickson ◽

Kenneth D Poss

Keyword(s):

Cardiac Injury ◽

Heart Regeneration ◽

Cardiomyocyte Proliferation ◽

Adult Zebrafish ◽

Perivascular Cells ◽

Adult Heart ◽

Muscle Hyperplasia ◽

Zebrafish Heart

Heart regeneration is limited in adult mammals but occurs naturally in adult zebrafish through the activation of cardiomyocyte division. Several components of the cardiac injury microenvironment have been identified, yet no factor on its own is known to stimulate overt myocardial hyperplasia in a mature, uninjured animal. In this study, we find evidence that Neuregulin1 (Nrg1), previously shown to have mitogenic effects on mammalian cardiomyocytes, is sharply induced in perivascular cells after injury to the adult zebrafish heart. Inhibition of Erbb2, an Nrg1 co-receptor, disrupts cardiomyocyte proliferation in response to injury, whereas myocardial Nrg1 overexpression enhances this proliferation. In uninjured zebrafish, the reactivation of Nrg1 expression induces cardiomyocyte dedifferentiation, overt muscle hyperplasia, epicardial activation, increased vascularization, and causes cardiomegaly through persistent addition of wall myocardium. Our findings identify Nrg1 as a potent, induced mitogen for the endogenous adult heart regeneration program.

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TCDD Inhibits Heart Regeneration in Adult Zebrafish

Toxicological Sciences ◽

10.1093/toxsci/kfs329 ◽

2012 ◽

Vol 132 (1) ◽

pp. 211-221 ◽

Cited By ~ 15

Author(s):

Peter Hofsteen ◽

Vatsal Mehta ◽

Min-Sik Kim ◽

Richard E. Peterson ◽

Warren Heideman

Keyword(s):

Heart Regeneration ◽

Adult Zebrafish

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Live imaging of angiogenesis during cutaneous wound healing in adult zebrafish

Angiogenesis ◽

10.1007/s10456-018-09660-y ◽

2019 ◽

Vol 22 (2) ◽

pp. 341-354 ◽

Cited By ~ 6

Author(s):

Chikage Noishiki ◽

Shinya Yuge ◽

Koji Ando ◽

Yuki Wakayama ◽

Naoki Mochizuki ◽

...

Keyword(s):

Wound Healing ◽

Live Imaging ◽

Cutaneous Wound Healing ◽

Adult Zebrafish ◽

Cutaneous Wound

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Developmental and Regenerative Biology of Cardiomyocytes

The International Journal of Developmental Biology ◽

10.1387/ijdb.210159kh ◽

2021 ◽

Author(s):

Angeliki Daiou ◽

Katerina Petalidou ◽

Georgios Siokatas ◽

Eleftherios I. Papadopoulos ◽

Konstantinos E. Hatzistergos

Keyword(s):

Developmental Biology ◽

Current Knowledge ◽

Heart Diseases ◽

Clinical Stage ◽

Critical Role ◽

Mouse Heart ◽

Heart Regeneration ◽

Adult Zebrafish ◽

Mammalian Embryo ◽

Metabolic Remodeling

The current progress and challenges in understanding the molecular and cellular mechanisms of cardiomyocyte embryonic development and regeneration are reviewed in our present work. Three major topics are critically discussed: how do cardiomyocytes form in the embryo? What is the adult origin of the cells that regenerate cardiomyocytes in animal models with adult heart regeneration capabilities? Can the promise of therapeutic cardiomyocyte regeneration be realized in humans? In the first topic, we highlight current advancements in understanding the developmental biology of cardiomyocytes, with emphasis on the regulative capabilities of the early embryo during specification and allocation of the cardiomyoblasts that produce the primordial heart. We further emphasize on trabecular cardiomyocyte development from late cardiomyoblasts, neural crest cells and primordial cardiomyocytes, and their critical role on the clonal growth of the compact/septal and cortical cardiomyocyte layers in the mammalian embryo and adult zebrafish, respectively. In the second topic, we focus on the reactivation of the cortical or trabecular compaction programs as hallmarks of cardiomyocyte regenerative cells during adult zebrafish and neonatal mouse heart regeneration, respectively, and underscore the metabolic remodeling that commonly drives cardiomyocyte regeneration in these organisms. Finally, we discuss the status of preclinical and clinical-stage therapeutics for cardiomyocyte regeneration, with particular emphasis on gene therapy, as well as adult and pluripotent stem cell-based cellular cardiomyoplasty approaches. In summary, our article provides a bird’s-eye view on the current knowledge and potential pitfalls in the field of developmental biology-guided regenerative medicine strategies for the treatment of heart diseases.

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