Abstract

The Mexican axolotl salamander (Ambystoma mexicanum) is one member of a select group of vertebrate animals that have retained the amazing ability to regenerate multiple body parts. In addition to being an important model system for regeneration, the axolotl has also contributed extensively to studies of basic development. While many genes known to play key roles during development have now been implicated in various forms of regeneration, much of the regulatory apparatus controlling the underlying molecular circuitry remains unknown. In recent years, microRNAs have been identified as key regulators of gene expression during development, in many diseases and also, increasingly, in regeneration. Here, we have used deep sequencing combined with qRT-PCR to undertake a comprehensive identification of microRNAs involved in regulating regeneration in the axolotl. Specifically, among the microRNAs that we have found to be expressed in axolotl tissues, we have identified 4564 microRNA families known to be widely conserved among vertebrates, as well as 59,811 reads of putative novel microRNAs. These findings support the hypothesis that microRNAs play key roles in managing the precise spatial and temporal patterns of gene expression that ensures the correct regeneration of missing tissues.

Highlights

  • Regeneration is defined as the regrowth or restoration of lost tissue

  • 100 bps) in regenerating tail blastema versus control mature tail tissue. This approach allowed us to identify conserved microRNA families and to identify putative novel microRNAs that we found especially enriched in the blastema sample

  • We validated expression of these known microRNA families using quantitative RT-PCR and in addition examined the dynamics of these microRNAs through the course of regeneration

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Summary

Introduction

Regeneration is defined as the regrowth or restoration of lost tissue. Mammals, have very limited regenerative wound healing capacities, addressing only small portions of liver, peripheral nerve damage and muscle through abilities that decline markedly with age. Salamanders, stand as the champions in this field, able to regenerate with full structural accuracy and functionality a long list of complex body parts including, but not limited to, limbs, tail, spinal cord, and portions of the heart [1–22] These remarkable regenerative capacities have been studied for many years and more recent technological advances have begun to enable the identification of underlying molecular effectors and pathways. Transcriptional profiling has revealed that many genes involved in basic limb and spinal cord development are reused, if and when those structures are regenerated [23,24] These studies confirmed that many of these developmental genes used by salamanders during regeneration show clear evolutionary conservation with those active during mammalian development. The fact that some of the effector genes involved are conserved in mammals without conferring fully functional regenerative abilities indicates that important differences remain elsewhere, begging a closer examination of upstream regulatory molecules and mechanisms

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