Abstract
BackgroundSalamanders regenerate their limbs after amputation. However, the molecular mechanism of this unique regeneration remains unclear. In this study, isobaric tags for relative and absolute quantification (iTRAQ) coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed to quantitatively identify differentially expressed proteins in regenerating limbs 3, 7, 14, 30 and 42 days post amputation (dpa).ResultsOf 2636 proteins detected in total, 253 proteins were differentially expressed during different regeneration stages. Among these proteins, Asporin, Cadherin-13, Keratin, Collagen alpha-1(XI) and Titin were down-regulated. CAPG, Coronin-1A, AnnexinA1, Cathepsin B were up-regulated compared with the control. The identified proteins were further analyzed to obtain information about their expression patterns and functions in limb regeneration. Functional analysis indicated that the differentially expressed proteins were associated with wound healing, immune response, cellular process, metabolism and binding.ConclusionsThis work indicated that significant proteome alternations occurred during salamander limb regeneration. The results may provide fundamental knowledge to understand the mechanism of limb regeneration.
Highlights
Histological observation of regenerating limbs in the longitudinal section showed that the wound stump was covered by epidermis and gland cells
The results indicated that significant proteomic alternations occurred during the process
The differentially expressed proteins (DEPs) detected may provide fundamental basis to understand the mechanism of limb regeneration
Summary
Isobaric tags for relative and absolute quantification (iTRAQ) coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed to quantitatively identify differentially expressed proteins in regenerating limbs 3, 7, 14, 30 and 42 days post amputation (dpa). The missing portion of a salamander limb could be perfectly regenerated after amputation between the shoulder and hand. Salamander limb regeneration could be divided into three major stages. Cells surrounding the injury site begin to dedifferentiate, proliferate and Regeneration of limbs is a complex process involving the activation of a number of biological processes, signaling pathways and large-scale tissue remodeling [12,13,14,15,16,17].
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