Faculty Opinions recommendation of Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo.

2021 ◽  
Author(s):  
Kazu Kikuchi
Keyword(s):  
Ex Vivo ◽  
Live Imaging ◽  
Cardiomyocyte Proliferation ◽  
Adult Zebrafish

scholarly journals Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

Development ◽  
2021 ◽  
Author(s):  
Hessel Honkoop ◽  
Phong D. Nguyen ◽  
Veronique E.M. van der Velden ◽  
Katharina F. Sonnen ◽  
Jeroen Bakkers
Keyword(s):  
Culture System ◽  
Ex Vivo ◽  
Live Imaging ◽  
Slice Culture ◽  
Heart Regeneration ◽  
Dependent Manner ◽  
Cardiomyocyte Proliferation ◽  
Adult Zebrafish ◽  
Cellular Mechanisms ◽  
Zebrafish Heart

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.

scholarly journals Cell shape anisotropy and motility constrain self-organised feather pattern fidelity in birds

2021 ◽  
Author(s):  
Camille Curantz ◽  
Richard Bailleul ◽  
Magdalena Hidalgo ◽  
Melina Durande ◽  
François Graner ◽  
...  
Keyword(s):  
Cell Motility ◽  
Cell Shape ◽  
Species Differences ◽  
Ex Vivo ◽  
Live Imaging ◽  
Developmental Pattern ◽  
Stochastic Fluctuations ◽  
Dermal Cell ◽  
Self Organisation ◽  
Cell Anisotropy

SummaryCellular self-organisation can emerge from stochastic fluctuations in properties of a developing tissue1–3. This mechanism explains the production of various motifs seen in nature4–7. However, events channelling its outcomes such that patterns are produced with reproducible precision key to fitness remain unexplored. Here, we compared the dynamic emergence of feather primordia arrays in poultry, finch, emu, ostrich and penguin embryos and correlated inter-species differences in pattern fidelity to the amplitude of dermal cell anisotropy in the un-patterned tissue. Using live imaging and ex vivo perturbations in these species, we showed that cell anisotropy optimises cell motility for sharp and precisely located primordia formation, and thus, proper pattern geometry. These results evidence a mechanism through which collective cellular properties of a developmental pattern system ensure stability in its self-organisation and contribute to its evolution.

Abstract 403: Leveraging the Zebrafish to Overcome Barriers in Heart Transplantation

2020 ◽  
Vol 127 (Suppl_1) ◽  
Author(s):  
Shannon N Tessier ◽  
Luciana Da Silveira Cavalcante ◽  
Casie A Pendexter ◽  
Stephanie E Cronin ◽  
Reinier J de Vries ◽  
...  
Keyword(s):  
Heart Transplantation ◽  
Ischemia Reperfusion Injury ◽  
Ex Vivo ◽  
Ischemia Reperfusion ◽  
Complex Structure ◽  
Scale Up ◽  
Adult Zebrafish ◽  
Zebrafish Larvae ◽  
Preservation Method ◽  
Heart Preservation

Cardiac transplantation is the only curative therapy for patients with end-stage heart disease; however, there is a severe shortage of viable donor organs. Heart transplantation faces many interwoven challenges, including both biological factors and research limitations. For example, ischemia-reperfusion injury plays a role in early graft dysfunction and is associated with rejection episodes in heart transplantation. Moreover, experimental transplantation relies heavily on animal studies that are laborious and expensive, prohibiting the discovery of novel, bold solutions. We propose that the zebrafish, Danio rerio , would be a valuable tool for the field since it’s amenable to high-throughput screens, captures the complex structure of organs, and offers a suite of tools to monitor the biology of cardiac injury. Here, we develop a new subzero heart preservation method by strategically leveraging animal models from zebrafish to mammalian hearts. Using zebrafish larvae, we screened for agents which preserve hearts at -10°C. As a result of these screens, we identified promising preservative cocktails which restored heartbeat in 82% of larvae immediately post-recovery. Next, we excised adult zebrafish hearts and developed methods to mimic the ex vivo handling practices of hearts destined for transplant using a heart-on-a-plate assay. Using this assay, we carried forward promising agents identified in our initial zebrafish larvae screen to isolated adult zebrafish hearts that were cooled to -10°C and held for up to 24 hours. After rewarming, heart rate was restored and metabolic rate of zebrafish hearts was like time-matched controls (0.213 ± 0.047 and 0.275 ± 0.060, respectively, p = 0.200). Finally, we report our preliminary scale-up efforts whereby rodent hearts are stored for up to 24 hours at -10°C and viability were assessed by the TUNEL assay. The data shows high viability of cardiomyocytes post-preservation, as compared to controls. In summary, we present data to illustrate our efforts in leveraging the zebrafish to aid new discoveries in subzero heart preservation. Similar efforts to model heart transplantation in zebrafish may provide a different vantage point and enable us to make advances faster.

scholarly journals Investigating the utility of adult zebrafish ex vivo whole hearts to pharmacologically screen hERG channel activator compounds

2019 ◽  
Vol 317 (6) ◽  
pp. R921-R931
Author(s):  
Christina M. Hull ◽  
Christine E. Genge ◽  
Yuki Hobbs ◽  
Kaveh Rayani ◽  
Eric Lin ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Expression System ◽  
Cost Effective ◽  
Channel Blockers ◽  
Loss Of Function ◽  
Adult Zebrafish ◽  
Pharmacological Characterization ◽  
Potential Gain ◽  
Xenopus Oocyte Expression ◽  
Whole Heart

There is significant interest in the potential utility of small-molecule activator compounds to mitigate cardiac arrhythmia caused by loss of function of hERG1a voltage-gated potassium channels. Zebrafish ( Danio rerio) have been proposed as a cost-effective, high-throughput drug-screening model to identify compounds that cause hERG1a dysfunction. However, there are no reports on the effects of hERG1a activator compounds in zebrafish and consequently on the utility of the model to screen for potential gain-of-function therapeutics. Here, we examined the effects of hERG1a blocker and types 1 and 2 activator compounds on isolated zkcnh6a (zERG3) channels in the Xenopus oocyte expression system as well as action potentials recorded from ex vivo adult zebrafish whole hearts using optical mapping. Our functional data from isolated zkcnh6a channels show that under the conditions tested, these channels are blocked by hERG1a channel blockers (dofetilide and terfenadine), and activated by type 1 (RPR260243) and type 2 (NS1643, PD-118057) hERG1a activators with higher affinity than hKCNH2a channels (except NS1643), with differences accounted for by different biophysical properties in the two channels. In ex vivo zebrafish whole hearts, two of the three hERG1a activators examined caused abbreviation of the action potential duration (APD), whereas hERG1a blockers caused APD prolongation. These data represent, to our knowledge, the first pharmacological characterization of isolated zkcnh6a channels and the first assessment of hERG enhancing therapeutics in zebrafish. Our findings lead us to suggest that the zebrafish ex vivo whole heart model serves as a valuable tool in the screening of hKCNH2a blocker and activator compounds.

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

Lab on a Chip ◽  
2020 ◽  
Vol 20 (2) ◽  
pp. 274-284
Author(s):  
Joycelyn K. Yip ◽  
Michael Harrison ◽  
Jessi Villafuerte ◽  
G. Esteban Fernandez ◽  
Andrew P. Petersen ◽  
...  
Keyword(s):  
Live Imaging ◽  
Heart Regeneration ◽  
Adult Zebrafish ◽  
Extended Culture ◽  
Fluidic Device

Culturing adult zebrafish hearts in a fluidic device reduces morphological and functional declines and enables live imaging of heart regeneration.

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

eLife ◽  
2015 ◽  
Vol 4 ◽  
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.

scholarly journals Progestin increases the expression of gonadotropins in pituitaries of male zebrafish

2016 ◽  
Vol 230 (1) ◽  
pp. 143-156 ◽  
Author(s):  
Cuili Wang ◽  
Dongteng Liu ◽  
Weiting Chen ◽  
Wei Ge ◽  
Wanshu Hong ◽  
...  
Keyword(s):  
Time Course ◽  
Ex Vivo ◽  
Mrna Levels ◽  
Adult Zebrafish ◽  
Male Adult ◽  
Positive Effects ◽  
Time Course Experiment ◽  
Nuclear Progesterone Receptor

Our previous study showed that the in vivo positive effects of 17α,20β-dihydroxy-4-pregnen-3-one (DHP), the major progestin in zebrafish, on early spermatogenesis was much stronger than the ex vivo ones, which may suggest an effect of DHP on the expression of gonadotropins. In our present study, we first observed that fshb and lhb mRNA levels in the pituitary of male adult zebrafish were greatly inhibited by 3 weeks exposure to 10nM estradiol (E2). However, an additional 24h 100nM DHP exposure not only reversed the E2-induced inhibition, but also significantly increased the expression of fshb and lhb mRNA. These stimulatory effects were also observed in male adult fish without E2 pretreatment, and a time course experiment showed that it took 24h for fshb and 12h for lhb to respond significantly. Because these stimulatory activities were partially antagonized by a nuclear progesterone receptor (Pgr) antagonist mifepristone, we generated a Pgr-knockout (pgr–/–) model using the TALEN technique. With and without DHP in vivo treatment, fshb and lhb mRNA levels of pgr–/– were significantly lower than those of pgr+/+. Furthermore, ex vivo treatment of pituitary fragments of pgr–/– with DHP stimulated lhb, but not fshb mRNA expression. Results from double-colored fluorescent in situ hybridization showed that pgr mRNA was expressed only in fshb-expressing cells. Taken together, our results indicated that DHP participated in the regulation of neuroendocrine control of reproduction in male zebrafish, and exerted a Pgr-mediated direct stimulatory effect on fshb mRNA at pituitary level.

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