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The essential role of connective-tissue cells during axolotl limb regeneration [Agilent]

ABSTRACT: This experiment was performed to obtain an expression profiling of connective-tissue-derived cells at different time points during blastema formation after limb amputation. RNA extraction was performed from FACS sorted connective tissue cells from converted Prrx1Cre-ERT;CaggsSTOP/Cherry axolotls. 3 Biological replicates were obtained for each time point analyzed.

ORGANISM(S): Ambystoma mexicanum

PROVIDER: GSE294346 | GEO | 2025/04/17

REPOSITORIES: GEO

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The essential role of connective-tissue cells during axolotl limb regeneration

Project description:Axolotls (Ambystoma mexicanum) are known for their remarkable limb-regeneration abilities, which involve the formation of the blastema, a specialized structure consisting of progenitor cells contributed by all major tissues of the limb. Lateral plate mesoderm (LPM)-derived connective tissue (CT) cells dedifferentiate and play a critical role in blastema formation and subsequent limb regeneration. However, the complexity of the blastema’s cellular composition and the extent of CT participation and necessity have not been rigorously explored. To address this gap, we conducted 10x Xenium spatial transcriptomics using a selected array of probes and finding the cellular identity and molecular architecture of blastema during the first stages of limb regeneration

2025-04-16 | GSE294212 | GEO

Comparison of Protein Expression During Early Phases of Limb Regeneration Between Regeneration-permissive and Regeneration-deficient Axolotl

Project description:In this project, we studied the proteomic profiles of the early stages of blastema formation of neotenic and metamorphic axolotls after limb amputation by means of LC-MS/MS technology. We quantified a total of 714 proteins having an adjusted p < 0.01 with FC greater or equal to 2 between two conditions. Principal component analysis revealed a conspicuous clustering between neotenic and metamorphic samples at 7 days post-amputation. Different set of proteins was identified as differentially expressed at all of the time points (1, 4, and 7 days post-amputations against day0) for neotenic and metamorphic stages. Although functional enrichment analyses underline the presence of common pathways between regenerative and non-regenerative stages, cell proliferation and its regulation associated pathways, immune system related pathways and muscle tissue and ECM remodeling and degradation pathways were represented at different rate between both stages.

2019-11-25 | PXD014806 | Pride

Three Distinct Phases of Regeneration-Specific Gene Expression in the Axolotl Blastema

Project description:The salamander has the remarkable ability to regenerate its limb after amputation. Cells at the site of amputation form a blastema and then proliferate and differentiate to regrow the limb. To better understand this process, we have performed deep RNA sequencing of the blastema over a time course. We find genes expressed in three phases with a prominent burst in oncogene expression during the first day, blastemal/limb bud genes peaking at 7 to 14 days, and markers for terminal differentiation upregulated later. We compare these expression patterns to those in a mouse digit amputation model to identify genes specific to the regenerative response. We find that limb patterning genes, SALL genes, and genes involved in the proteasome, adult stem cell, embryonic stem cell, retinoid metabolism, and WNT and NOTCH signaling are regeneration-specific. We establish the “Axolomics” Database, which provides a tool for depositing, retrieving, and searching axolotl-related “omic” information. The experiment includes two time course data sets. One is from mouse digit amputation, another is from Axolotl digit amputation

2013-09-01 | E-GEOD-34394 | biostudies-arrayexpress

Single-cell analysis uncovers convergence of cell identities during axolotl limb regeneration

Project description:Amputation of the axolotl forelimb results in the formation of a blastema, a transient tissue where progenitor cells accumulate prior to limb regeneration. Connective tissue (CT) – skeleton, periskeleton, tendon, dermis, interstitial cells – contributes the vast majority of cells that populate the blastema, however it is unclear how individual CT cells may reprogram their fate in order to rebuild the tetrapod limb. Here we use a combination of Cre-loxP reporter lineage tracking and single-cell (sc) RNA-seq to molecularly track, for the first time, adult CT cell heterogeneity and its transition to a limb blastema state. We uncover a multi-phasic molecular program where CT cell types found in the uninjured adult limb revert to a relatively homogenous progenitor state that participates in inflammation and extracellular matrix disassembly prior to proliferation, establishment of positional information, and ultimately re-differentiation. While the early regeneration transcriptome states are unique to the blastema, the later stages recapitulate embryonic limb development. Notably, we do not find evidence of a pre-existing blastema-like precursor nor limb bud-like progenitors in the uninjured adult tissue. However, we find that distinct CT subpopulations in the adult limb differentially contribute to proximal and distal portions of the regenerated limb. Together, our data illuminates molecular and cellular reprogramming during complex organ regeneration in a vertebrate.

2018-09-27 | GSE106269 | GEO

Three Distinct Phases of Regeneration-Specific Gene Expression in the Axolotl Blastema

2013-09-01 | GSE34394 | GEO

The essential role of connective-tissue cells during axolotl limb regeneration [Agilent]

Project description:The essential role of connective-tissue cells during axolotl limb regeneration [Agilent]

| PRJNA1249071 | ENA

Transcriptomics of A. mexicanum Regeneration: A Comparative Study Reveals Key Proteins And Pathways for Limb Regeneration.

Project description:The axolotl is an animal with remarkable regenerative abilities, making it an ideal model for studying potential regenerative therapies in mammals, including humans. However, the molecular mechanisms involved in regeneration remain unclear. We conducted a transcriptomic analysis of juvenile axolotls' limbs and their blastema and compared the results with aged axolotls that failed to regenerate after amputation. We identified a set of genes involved in cell differentiation, transcriptional regulation, cartilage development, bone morphogenesis, and extracellular matrix remodeling. Four highly expressed genes (FSTL1, ADAMTS17, GPX7, and CTHRC1) were identified in regenerating tissue, but underexpressed in aged axolotls. Structural and homology analysis showed that these genes are conserved and have important roles in development, bone morphogenesis, and cartilage formation. Our findings propose a novel set of key axolotl genes involved in tissue regeneration that could be a starting point for further studies in other vertebrates.

2024-11-27 | GSE237864 | GEO

Evi5 is required for Xenopus limb and tail regeneration

Project description:Amphibians such as the salamanders and the African clawed frog Xenopus are great models for regeneration studies because they can fully regenerate their lost organs. While axolotl can regenerate damaged organs throughout its lifetime, Xenopus has a limited regeneration capacity after metamorphosis. The ecotropic viral integrative factor 5 (Evi5), a cell-cycle-regulated protein that prevents cells from entering mitosis prematurely, is of great interest for it is highly upregulated in the limb blastema of axolotls, but its expression level remains unchanged in the fibroblastema of postmetamorphic frogs. Yet, its role in regeneration competent context in Xenopus has not been fully analyzed. Here we show that Evi5 is also upregulated in Xenopus tadpoles after limb and tail amputation, as it is in axolotls. Down-regulation of Evi5 with morpholino antisense oligos (Mo) impairs wound healing and blastema formation in limbs and tails in both axolotls and Xenopus tadpoles, suggesting a conserved function for Evi5 in regeneration. Using skin punch as a healing model we show that Evi5 is also involved in cell migration during wound healing. RNA-sequencing analysis shows that in addition to reduced signaling of Lepr, Pdgfa, Gdf5, evi5 Mo also downregulate lysine demethylases kdm6b and kdm7a, which are also required for limb regeneration. Thus, our results demonstrate that Evi5 plays a critical role in the regeneration of multiple systems in amphibians.

2022-11-24 | GSE218034 | GEO

Adrenergic signaling stimulates body-wide stem cell activation for limb regeneration (2N RNA-Seq)

Project description:In axolotls, stem cells are systemically prompted to re-enter the cell cycle upon amputation. We sought to identify how the cells surrounding the systemically activated cells respond to an amputation.

2023-05-13 | GSE232082 | GEO

Adrenergic signaling stimulates body-wide stem cell activation for limb regeneration (4N RNA-Seq)

Project description:In axolotls, stem cells are systemically prompted to re-enter the cell cycle upon amputation. We sought to identify which cells drive systemic-activation response in the limbs distant to the amputation.

2023-05-13 | GSE232083 | GEO

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