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Antler Proteome Atlas for endochondral ossification

ABSTRACT: Antler primary growth center is located in the distal tip of growing antler with five consecutive tissue layers under the velvet skin. Because of blastema progenitor cells of outmost reserve mesenchymal layer with multipotent differentiation capacity, antler regeneration recapitulates embryonic skeletal development through endochondral os-sification process. To further decipher this modified endochondral ossification, we profile deer antler proteome atlas with 8000 proteins, covering the whole five layers that comprehensively represent the mesenchymal condensation, rapid mesenchymal proliferation, chondrocyte hypertrophy, cartilage mineralization. By integrating three clustering methods including WGCNA, Fuzzy C-means clustering, and Monotonic feature selector, we elucidate proteome dynamics within the growth center, identifying key factors and elucidating biological processes through GO enrichment analysis. Through comparative analysis of RNA-seq and Proteomic studies, our results reveal that mRNA exhibits greater fluctuations during the endochondral ossification process. This research offers a valuable model for investigating hub genes and their biological function involved in organ regeneration.

ORGANISM(S): Cervus Nippon Mantchuricus

SUBMITTER: Xiangyan Li

PROVIDER: PXD066352 | iProX | Mon Jul 21 00:00:00 BST 2025

REPOSITORIES: iProX

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Project description:As the only known mammalian organ that can fully and annually regenerate, deer antler has significant advantages over lower-order animal models when investigating the control of stem cell-based organ regeneration. Antler regeneration is known to be initiated and maintained by neural crest-derived stem cells in different states of activation. Antler stem cells can therefore be used as a model system to study the proteins and pathways involved in the maintenance of a stem cell niche and their activation and differentiation during organ formation. In the current study, the MSC markers CD73, CD90 and CD105 were examined within the antler tip. Label-free quantification was performed to investigate the protein profiles of antler stem cells under different stages of activation and included: dormant pedicle periosteum (DPP), antler growth center (GC), post-active stem cells from mid-beam antler periosteum (MAP), and deer facial periosteum (FP) as a control (n = 3 per group). PEAKS and IPA software were used to analyze the proteomic data. Our research confirmed the central role of stem cell activation in the development of this mammalian organ by localizing the MSC markers within the antler growth center. Label-free quantification revealed that the greatest number of unique proteins (eighty-seven) was found in the growth center tissue. There were only 12 proteins found with expression levels that significantly differed between DPP and FP. Protein profiles of these two groups indicated that antler stem cells may use similar mechanisms to maintain dormancy within a stem cell niche. The number of significantly regulated proteins between DPP, MAP and GC was 153. Among them, the majority were upregulated in the growth center. Activation of antler stem cells was associated with a number of biological processes and signaling pathways such as Hippo and canonical Wnt signaling. This work identifies the key canonical pathways, molecular/cellular functions and upstream regulators involved in mammal organ regeneration.

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Antler Proteome Atlas for endochondral ossification

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