Identification of cDNAs associated with late dedifferentiation in adult newt forelimb regeneration

Abstract

Epimorphic limb regeneration in the adult newt involves the dedifferentiation of differentiated cells to yield a pluripotent blastemal cell. These mesenchymal-like cells proliferate and subsequently respond to patterning and differentiation cues to form a new limb. Understanding the dedifferentiation process requires the selective identification of dedifferentiating cells within the heterogeneous population of cells in the regenerate. In this study, representational differences analysis was used to produce an enriched population of dedifferentiation-associated cDNA fragments. Fifty-nine unique cDNA fragments were identified, sequenced, and analyzed using bioinformatics tools and databases. Some of these clones demonstrate significant similarity to known genes in other species. Other clones can be linked by homology to pathways previously implicated in the dedifferentiation process. These data will form the basis for further analyses to elucidate the role of candidate genes in the dedifferentiation process during newt forelimb regeneration. Developmental Dynamics 233:347-355, 2005.

Mechanisms of urodele limb regeneration

“…Microarray and RNA‐Seq analysis of regenerating axolotl limbs has identified suites of genes encoding progenitor cell markers, stage‐specific genes, and genes regulated by neural signals (Knapp et al., 2013; Looso et al., 2013; Mercer et al., 2012; Monaghan et al., 2009, 2012; Stewart et al., 2013; Vascotto, Beug, Liversage, & Tsilfildis, 2005; Voss et al., 2015). Bryant, et al.…”

Gene Expression Profile in the Anterior Regeneration of the Earthworm Using Expressed Sequence Tags

“…6) Additionally, cell-cell communication and biosynthesis actively take place during the early stages of regeneration to induce de-differentiation, to regulate the proliferation of pluripotent cells, and to respecify the fates of such cells to reconstruct the missing organs. 7) Therefore, analysis of transcripts in the early stage of anterior regeneration of the earthworm might provide a unique and valuable data set for understanding the molecular mechanism of the loss and gaining of cell identity that occurs during anterior regeneration, and to identify the candidate molecules responsible for controlling the de-differentiation and redifferentiation of the pivotal system, which includes the central nervous system.…”

Newt Opportunities for Understanding the Dedifferentiation Process

“…Vascotto et al conducted representational difference analysis (RDA) in order to identify genes associated with the dedifferentiation process. This screen isolated cDNAs from several gene classes, including putative transcription factors, immune modulators, proteases, and genes involved in cytoskeletal remodeling [50]. The homeobox gene msx-1 has also been shown to have a key role in dedifferentiation [51,52] and will be discussed further below.…”