Intrathoracic Injection for the Study of Adult Zebrafish Heart

Bise, Thomas; Jaźwińska, Anna

doi:10.3791/59724

Cited by 10 publications

(12 citation statements)

References 36 publications

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“…Clodronate liposomes (5 μl at a concentration of 10 mg/ml, Cat# F70101C‐A, FormuMax Scientific, Inc.), Dexamethasone (Bollaerts et al , 2019) (3 μl at a concentration of 1.65 mg/ml in sterile 0.68% saline; Cat# D1756, Sigma‐Aldrich), or PBS was injected intrathoracically 1 day before apex resection into individual anaesthetized zebrafish using a Picospritzer microinjector (Parker) as described previously(Bevan et al , 2019; Bise & Jazwinska, 2019). The injection was repeated one more time at 3 dpi.…”

Section: Methodsmentioning

confidence: 99%

Functional coordination of non‐myocytes plays a key role in adult zebrafish heart regeneration

Ma¹,

Liu²,

Yang

et al. 2021

EMBO Reports

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Cardiac regeneration occurs primarily through proliferation of existing cardiomyocytes, but also involves complex interactions between distinct cardiac cell types including non-cardiomyocytes (non-CMs). However, the subpopulations, distinguishing molecular features, cellular functions, and intercellular interactions of non-CMs in heart regeneration remain largely unexplored. Using the LIGER algorithm, we assemble an atlas of cell states from 61,977 individual non-CM scRNA-seq profiles isolated at multiple time points during regeneration. This analysis reveals extensive non-CM cell diversity, including multiple macrophage (MC), fibroblast (FB), and endothelial cell (EC) subpopulations with unique spatiotemporal distributions, and suggests an important role for MC in inducing the activated FB and EC subpopulations. Indeed, pharmacological perturbation of MC function compromises the induction of the unique FB and EC subpopulations. Furthermore, we developed computational algorithm Topologizer to map the topological relationships and dynamic transitions between functional states. We uncover dynamic transitions between MC functional states and identify factors involved in mRNA processing and transcriptional regulation associated with the transition. Together, our single-cell transcriptomic analysis of non-CMs during cardiac regeneration provides a blueprint for interrogating the molecular and cellular basis of this process.

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Section: Methodsmentioning

confidence: 99%

Functional coordination of non‐myocytes plays a key role in adult zebrafish heart regeneration

Ma¹,

Liu²,

Yang

et al. 2021

EMBO Reports

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“…Fish were closely monitored for the first 2 hours after surgery and twice daily thereafter until ready for organ collection. Pharmacological inhibition of oxytocin signaling was achieved by injecting 2-3 μL of water or 1 μM L-368,899 (a non-peptide OXTR antagonist) directly into the intrathoracic cavity, as described previously (Bise & Jaźwińska, 2019). Initial injections were administered 24 hours after cryoinjury with repeat injections every other day throughout the experiment.…”

Section: Methodsmentioning

confidence: 99%

Oxytocin promotes epicardial cell activation and heart regeneration after cardiac injury

Wasserman

Lewis-Israeli

Huang

et al. 2021

Preprint

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Cardiovascular disease (CVD) is one of the leading causes of mortality worldwide, and frequently leads to massive heart injury and the loss of billions of cardiac muscle cells and associated vasculature. Critical work in the last two decades demonstrated that these lost cells can be partially regenerated by the epicardium, the outermost mesothelial layer of the heart, in a process that highly recapitulates its role in heart development. Upon cardiac injury, mature epicardial cells activate and undergo an epithelial-mesenchymal transition (EMT) to form epicardial-derived progenitor cells (EpiPCs), multipotent progenitors that can differentiate into several important cardiac lineages, including cardiomyocytes and vascular cells. In mammals, this process alone is insufficient for significant regeneration, but it may be possible to prime it by administering specific reprogramming factors, leading to enhanced EpiPC function. Here, we show that oxytocin (OXT), a hypothalamic neuroendocrine peptide, induces epicardial cell proliferation, EMT, and migration in a mature-like model of human induced pluripotent stem cell (hiPSC)-derived epicardial cells. In addition, we demonstrate that OXT is released from the brain into the bloodstream after cardiac cryoinjury in zebrafish, eliciting significant epicardial activation and promoting heart regeneration. Oxytocin signaling is also critical for proper epicardium and myocardium development in zebrafish embryos. The above processes are significantly impaired when OXT signaling is inhibited chemically and genetically through RNA interference. Mechanistically, RNA sequencing analyses suggest that the transforming growth factor beta (TGF-β) pathway is the primary mediator of OXT-induced epicardial activation. Our research reveals for the first time a primarily brain-controlled mechanism that induces cellular reprogramming and regeneration of the injured heart, a finding that could yield significant translational advances for the treatment of CVD.

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“…To collect the heart for fixation, fish were euthanized in 0.6 mM tricaine solution. An incision was made above the heart through the branchial cartilage and the heart was pulled from the body cavity as shown in the video protocol 54 .…”

Section: Methodsmentioning

confidence: 99%

Multiple cryoinjuries modulate the efficiency of zebrafish heart regeneration

Bise

Sallin

Pfefferli

et al. 2020

Sci Rep

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Zebrafish can regenerate their damaged hearts throughout their lifespan. It is, however, unknown, whether regeneration remains effective when challenged with successive cycles of cardiac damage in the same animals. Here, we assessed ventricular restoration after two, three and six cryoinjuries interspaced by recovery periods. Using transgenic cell-lineage tracing analysis, we demonstrated that the second cryoinjury damages the regenerated area from the preceding injury, validating the experimental approach. We identified that after multiple cryoinjuries, all hearts regrow a thickened myocardium, similarly to hearts after one cryoinjury. However, the efficiency of scar resorption decreased with the number of repeated cryoinjuries. After six cryoinjuries, all examined hearts failed to completely resolve the fibrotic tissue, demonstrating reduced myocardial restoration. This phenotype was associated with enhanced recruitment of neutrophils and decreased cardiomyocyte proliferation and dedifferentiation at the early regenerative phase. Furthermore, we found that each repeated cryoinjury increased the accumulation of collagen at the injury site. Our analysis demonstrates that the cardiac regenerative program can be successfully activated many times, despite a persisting scar in the wounded area. This finding provides a new perspective for regenerative therapies, aiming in stimulation of organ regeneration in the presence of fibrotic tissue in mammalian models and humans.

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Intrathoracic Injection for the Study of Adult Zebrafish Heart

Cited by 10 publications

References 36 publications

Functional coordination of non‐myocytes plays a key role in adult zebrafish heart regeneration

Functional coordination of non‐myocytes plays a key role in adult zebrafish heart regeneration

Oxytocin promotes epicardial cell activation and heart regeneration after cardiac injury

Multiple cryoinjuries modulate the efficiency of zebrafish heart regeneration

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