The cellular and molecular mechanisms of tissue repair and regeneration as revealed by studies in <i>Xenopus</i>

Li, Jingjing; Zhang, Siwei; Amaya, Enrique

doi:10.1002/reg2.69

Cited by 32 publications

(25 citation statements)

References 92 publications

(147 reference statements)

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“…Of the three, mechanical wounding and observation (Steps 1-6) is the most accessible. When combined with gene knockdown/knockout or chemical treatment, it provides a rapid and robust assessment whether a certain gene, protein, or signaling pathway is involved in wound healing (Li et al , 2016Soto et al 2013). Nonetheless, it does require some practice to make the size and depth of wounds consistent, which can affect the speed and quality of healing.…”

Section: Discussionmentioning

confidence: 99%

Investigating the Cellular and Molecular Mechanisms of Wound Healing in Xenopus Oocytes and Embryos

Amaya

2018

Cold Spring Harb Protoc

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The African clawed frog Xenopus has remarkable capacities to heal wounds rapidly and to regenerate complex tissues. Because of its experimental tractability, studies using Xenopus oocytes, embryos, and larvae have contributed extensively to our understanding of the molecular and cellular mechanisms underpinning wound healing and tissue regeneration. In this protocol, we describe wound-healing assays following mechanical or laser injuries of oocytes and multicellular epithelia in Xenopus laevis embryos. We also explain how to perform assays aimed at investigating the cellular and molecular events during wound healing, including gene knockdown and overexpression experiments. In the latter assays, we explore the use of biochemical pull-down assays to investigate the activity of Rho GTPases, as well as the injection of mRNAs encoding fluorescent proteins or probes, followed by quantitative confocal image analyses to assay the dynamics of cytoskeletal components and their regulators. MATERIALSIt is essential that you consult the appropriate Material Safety Data Sheets and your institution's Environmental Health and Safety Office for proper handling of equipment and hazardous materials used in this protocol. RECIPES: Please see the end of this protocol for recipes indicated by . Additional recipes can be found online at http://cshprotocols.cshlp.org/site/recipes. ReagentsAgarose Antibodies, anti-Cdc42 (Cell Signaling Technology 2462) (for GST pull-down assays only) Antibodies, anti-Rac1/2/3 (Cell Signaling Technology 2465) (for GST pull-down assays only) Antibodies, anti-RhoA (Santa Cruz Biotechnology SC-179) (for GST pull-down assays only) Bovine serum albumin (BSA) Constructs (optional; see Steps 1, 7.ii, 7.iv, 11) Embryos can be injected with moesin-gfp to observe actin dynamics ), gfp-α-tubulin to observe microtubule dynamics (Woolner and Papalopulu 2012), GEM-GECO or C2-mrfp to observe calcium dynamics (Clark et al. 2009; Soto et al. 2013), or gfp/mcherry-caax to label the plasma membrane and thus outline the cell 3 Correspondence: enrique.amaya@manchester.ac.uk From the Xenopus collection, edited by Hazel L. Sive.

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Section: Discussionmentioning

confidence: 99%

Investigating the Cellular and Molecular Mechanisms of Wound Healing in Xenopus Oocytes and Embryos

Amaya

2018

Cold Spring Harb Protoc

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“…In summary, upon injury, TGF-b signaling mediated by smad2/3 is required for formation of the wound epidermis [9]. Extracellular matrix (ECM) remodeling, reactive oxygen species (ROS) signaling and inflammation are also among the first responses to tail injury [10]. Transcriptomic analysis of genes upregulated in the tail AEC identified pan-AEC factors that act both in regenerating limbs and tails, as well as two tail-specific AEC factors, Angptl2 and Egfl6 [11].…”

Section: Introductionmentioning

confidence: 99%

“…ROCs express genes from pathways known to be necessary for regeneration, such as the Wnt and Fgf pathways. As the regeneration bud forms and tail outgrowth occurs, additional signaling pathways are deployed, such as Bmp, Egf, Shh, Tgf-b and Notch pathways, among others [3][4]10,13]. RNA-seq analysis of isolated proliferating blastema cells revealed blastema-enriched genes such as Il11, Cse1l and L1td1-like [14].…”

Section: Introductionmentioning

confidence: 99%

Salamander-like tail regeneration in the West African lungfish

Verissimo

Perez

Dragalzew

et al. 2020

Preprint

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Salamanders, frog tadpoles, and lizards possess the remarkable ability to regenerate tails. The fossil record suggests that this capacity is an ancestral tetrapod trait, yet its evolutionary history remains unclear. Here we examine tail regeneration in a living representative of the sister group of tetrapods, the West African lungfish Protopterus annectens. We show that, as seen in salamanders, lungfish tail regeneration occurs via formation of a proliferative blastema and restores original structures including muscle, skeleton and spinal cord. Contrary to lizards and similar to salamanders, lungfish regenerate spinal cord neurons and reconstitute dorsoventral patterning of the tail.Similar to salamander and frog tadpoles, we show that Shh is required for lungfish tail regeneration. Through RNA-seq analysis of uninjured and regenerating tail blastema we show that lungfish deploy a genetic program comparable to that of tetrapods, showing upregulation of genes and signaling pathways previously implicated in amphibian and lizard tail regeneration. Furthermore, the tail blastema showed marked upregulation of genes encoding post-transcriptional RNA processing components and transposon-derived genes. Collectively, our study establishes the lungfish as a valuable research system for regenerative biology and provides insights into the evolution of cellular and molecular processes underlying vertebrate tail regeneration.

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“…What makes amphibian limb regeneration one of the most fascinating processes in nature is that the amphibian limb stump mobilizes cells at the amputation surface to form a regeneration blastema, from which a new limb is regrown . In recent decades, Xenopus has become a favorable regeneration model because of its unique regeneration behavior . Xenopus tadpoles can fully regenerate limbs during the period of limb outgrowth and differentiation before NF stage 54 10 (about 4 weeks old at 23 °C).…”

mentioning

confidence: 99%

“…[1][2][3][4] In recent decades, Xenopus has become a favorable regeneration model because of its unique regeneration behavior. [5][6][7][8][9] Xenopus tadpoles can fully regenerate limbs during the period of limb outgrowth and differentiation before NF stage 54 10 (about 4 weeks old at 23 8C). This regeneration ability declines gradually, in a proximal to distal wave concurring with the proximal-distal wave of differentiation of the limb, and is lost during metamorphosis.…”

mentioning

confidence: 99%

Dicer inactivation stimulates limb regeneration ability in Xenopus laevis

Zhang

Yang

Yuan

et al. 2018

Wound Repair Regeneration

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The ontogenetic decline of regeneration capacity in the anuran amphibian Xenopus makes it an excellent model for regeneration studies. However, the cause of the regeneration ability decline is not fully understood. MicroRNAs regulate animal development and have been indicated in various regeneration situations. However, little is known about the role of microRNAs during limb regeneration in Xenopus. This study investigates the effect of Dicer, an enzyme responsible for microRNA maturation, on limb development and regeneration in Xenopus. Dicer is expressed in the developing Xenopus limbs and is up-regulated after limb amputation during both regeneration-competent and regeneration-deficient stages of tadpole development. Inactivation of Dicer in early (NF stage 53) tadpole limb buds leads to shorter tibulare/fibulare formation but does not affect limb regeneration. However, in late-stage, regeneration-deficient tadpole limbs (NF stage 57), Dicer inactivation restores the regeneration blastema and stimulates limb regeneration. Thus, our results demonstrated that Xenopus limb regeneration can be stimulated by the inactivation of Dicer in nonregenerating tadpoles, indicating that microRNAs present in late-stage tadpole limbs may be involved in the ontogenetic decline of limb regeneration in Xenopus.

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The cellular and molecular mechanisms of tissue repair and regeneration as revealed by studies in Xenopus

Cited by 32 publications

References 92 publications

Investigating the Cellular and Molecular Mechanisms of Wound Healing in Xenopus Oocytes and Embryos

Investigating the Cellular and Molecular Mechanisms of Wound Healing in Xenopus Oocytes and Embryos

Salamander-like tail regeneration in the West African lungfish

Dicer inactivation stimulates limb regeneration ability in Xenopus laevis

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