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

Nandini Rao, Fengyu Song, Deepali Jhamb, Mu Wang, Derek J. Milner, Nathaniel M. Price, Teri L. Belecky-Adams, Mathew J. Palakal, Jo Ann Cameron, Bingbing Li, Xiaoping Chen, David L. Stocum

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish
Article number32
JournalBMC Developmental Biology
Volume14
Issue number1
DOIs
StatePublished - Jul 25 2014

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Ambystoma mexicanum
Xenopus laevis
Proteomics
Xenopus
Extremities
Hindlimb
Chondrocytes
Mitosis
Proteins
Down-Regulation
Virus Integration
Cell Cycle Proteins
Inositol Phosphates
Muscle Proteins
Extracellular Matrix Proteins
Diglycerides
Carbohydrate Metabolism
Amputation
Regeneration
Mass Spectrometry

Keywords

  • Comparison to axolotl
  • Fibroblastema formation
  • Proteomic analysis
  • Regeneration
  • Xenopus hindlimb

ASJC Scopus subject areas

  • Developmental Biology

Cite this

Proteomic analysis of fibroblastema formation in regenerating hind limbs of Xenopus laevis froglets and comparison to axolotl. / Rao, Nandini; Song, Fengyu; Jhamb, Deepali; Wang, Mu; Milner, Derek J.; Price, Nathaniel M.; Belecky-Adams, Teri L.; Palakal, Mathew J.; Cameron, Jo Ann; Li, Bingbing; Chen, Xiaoping; Stocum, David L.

In: BMC Developmental Biology, Vol. 14, No. 1, 32, 25.07.2014.

Research output: Contribution to journalArticle

Rao, N, Song, F, Jhamb, D, Wang, M, Milner, DJ, Price, NM, Belecky-Adams, TL, Palakal, MJ, Cameron, JA, Li, B, Chen, X & Stocum, DL 2014, 'Proteomic analysis of fibroblastema formation in regenerating hind limbs of Xenopus laevis froglets and comparison to axolotl', BMC Developmental Biology, vol. 14, no. 1, 32. https://doi.org/10.1186/1471-213X-14-32
Rao, Nandini ; Song, Fengyu ; Jhamb, Deepali ; Wang, Mu ; Milner, Derek J. ; Price, Nathaniel M. ; Belecky-Adams, Teri L. ; Palakal, Mathew J. ; Cameron, Jo Ann ; Li, Bingbing ; Chen, Xiaoping ; Stocum, David L. / Proteomic analysis of fibroblastema formation in regenerating hind limbs of Xenopus laevis froglets and comparison to axolotl. In: BMC Developmental Biology. 2014 ; Vol. 14, No. 1.
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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.",
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AU - Wang, Mu

AU - Milner, Derek J.

AU - Price, Nathaniel M.

AU - Belecky-Adams, Teri L.

AU - Palakal, Mathew J.

AU - Cameron, Jo Ann

AU - Li, Bingbing

AU - Chen, Xiaoping

AU - Stocum, David L.

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