Abstract
As the only known organ that can completely regenerate in mammals, deer antler is of real significance in the field of regenerative medicine. Recent studies have shown that the regenerative capacity of the antlers comes from the pedicle periosteum and the cells resident in the periosteum possess the attributes of stem cells. Currently, the molecular mechanism of antler regeneration remains unclear. In the present study, we compared the potentiated and dormant antler stem cells using isobaric tags for the relative and absolute quantification (iTRAQ) labeling of the peptides, coupled with two-dimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS) to compare the proteome profiles. Proteins were identified by searching against the NCBI nr database and our own Cervine transcriptome database, and bioinformatics analysis was conducted to identify the differentially expressed proteins. Based on this searching strategy, we identified 169 differentially expressed proteins in total, consisting of 70 up- and 99 down-regulated in the potentiated vs. dormant antler stem cells. Reliability of the iTRAQ was confirmed via quantitative real-time polymerase chain reaction (qRT-PCR) to measure the expression of selected genes. We identified transduction pathways through the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, such as HIF-1 and PI3K-AKT signaling pathways that play important roles in regulating the regeneration of antlers. In summary, the initiation stage of antler regeneration, a process from dormant to potentiated states in antler stem cells, is regulated by multiple proteins and complicated signal networks.
Highlights
Regenerative medicine is a field that aims to grow back damaged/lost tissues and organs via stimulation of the body’s own reparative capability [1]
Our results provided the first evidence, at the molecular level, that the potentiated antler stem cells express more proteins consistent with antler regeneration than the dormant antler stem cells, and lay the foundation for the eventual identification of the molecules that are involved in the initiation of antler regeneration
We obtained 169 regulated proteins in total from both the potentiated PP” (PPP) cells (PPPCs) and the dormant PP” (DPP) cells (DPPCs) based on the databases of both NCBI nr (Table S1) and our own (Table S2). Of these 169 proteins, 43 (19 + 24) proteins were unique to the NCBI nr database; 52 (26 + 26) proteins were unique to our own database; 74 (25 + 49) proteins were found in both databases (Figure 1)
Summary
Regenerative medicine is a field that aims to grow back damaged/lost tissues and organs via stimulation of the body’s own reparative capability [1]. Deer antlers are the only mammalian appendages that can achieve complete epimorphic regeneration, and this is initiated annually from the distal ends of the permanent bony protuberances on their head know as pedicles [3]. Evidence from both histology [4] and tissue deletion experiments [5] have convincingly demonstrated that it is the distal part of pedicle periosteum (PP) that gives rise to a regenerating antler. Further functional analysis [8] found that the distal third of the PP regenerated an antler; whereas, the proximal two thirds PP failed to do so after separating from the enveloping skin by insertion of an impermeable membrane All these results indicate that the mesenchymal/epithelial interactions between the PP and the enveloping skin are important for the initiation of antler regeneration. Identification of the differentially expressed proteins and the activated signaling pathways in the potentiated over the dormant antler stem cells would help to unravel the underlying molecular mechanism of antler regeneration
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