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Open Access Highly Accessed Research article

Proteomic analysis of fibroblastema formation in regenerating hind limbs of Xenopus laevis froglets and comparison to axolotl

Nandini Rao1, Fengyu Song2, Deepali Jhamb3, Mu Wang5, Derek J Milner9, Nathaniel M Price4, Teri L Belecky-Adams6, Mathew J Palakal8, Jo Ann Cameron9, Bingbing Li7, Xiaoping Chen4 and David L Stocum6*

Author Affiliations

1 Department of Biochemistry and Genetics, School of Medicine, American University of Antigua, Coolidge, Antigua, West Indies

2 Department of Oral Biology, School of Dentistry and Center for Developmental and Regenerative Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA

3 School of Informatics and Computing, Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA

4 Department of Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA

5 Department of Biochemistry and Molecular Biology, School of Medicine, and Center for Developmental and Regenerative Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA

6 Department of Biology, and Center for Developmental and Regenerative Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN, USA

7 Department of Chemistry, Central Michigan University, Mt. Pleasant, MI, USA

8 School of Informatics and Computing, and Center for Developmental and Regenerative Biology, Indiana University-Purdue University, Indianapolis, IN, USA

9 Department of Cell and Developmental Biology, and Regeneration Biology and Tissue Engineering Theme, Institute for Genomic Biology, University of Illinois-Urbana Champaign, Urbana, IL, USA

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BMC Developmental Biology 2014, 14:32  doi:10.1186/1471-213X-14-32

Published: 25 July 2014

Abstract

Background

To gain insight into what differences might restrict the capacity for limb regeneration in Xenopus froglets, we used High Performance Liquid Chromatography (HPLC)/double mass spectrometry to characterize protein expression during fibroblastema formation in the amputated froglet hindlimb, and compared the results to those obtained previously for blastema formation in the axolotl limb.

Results

Comparison of the Xenopus fibroblastema and axolotl blastema revealed several similarities and significant differences in proteomic profiles. The most significant similarity was the strong parallel down regulation of muscle proteins and enzymes involved in carbohydrate metabolism. Regenerating Xenopus limbs differed significantly from axolotl regenerating limbs in several ways: deficiency in the inositol phosphate/diacylglycerol signaling pathway, down regulation of Wnt signaling, up regulation of extracellular matrix (ECM) proteins and proteins involved in chondrocyte differentiation, lack of expression of a key cell cycle protein, ecotropic viral integration site 5 (EVI5), that blocks mitosis in the axolotl, and the expression of several patterning proteins not seen in the axolotl that may dorsalize the fibroblastema.

Conclusions

We have characterized global protein expression during fibroblastema formation after amputation of the Xenopus froglet hindlimb and identified several differences that lead to signaling deficiency, failure to retard mitosis, premature chondrocyte differentiation, and failure of dorsoventral axial asymmetry. These differences point to possible interventions to improve blastema formation and pattern formation in the froglet limb.

Keywords:
Regeneration; Xenopus hindlimb; Proteomic analysis; Fibroblastema formation; Comparison to axolotl