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Ba H, Wang X, Wang D, Ren J, Wang Z, Sun HX, Hu P, Zhang G, Wang S, Ma C, Wang Y, Wang E, Chen L, Liu T, Gu Y, Li C. Single-cell transcriptome reveals core cell populations and androgen-RXFP2 axis involved in deer antler full regeneration. CELL REGENERATION (LONDON, ENGLAND) 2022; 11:43. [PMID: 36542206 PMCID: PMC9772379 DOI: 10.1186/s13619-022-00153-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 05/29/2022] [Accepted: 11/11/2022] [Indexed: 12/24/2022]
Abstract
Deer antlers constitute a unique mammalian model for the study of both organ formation in postnatal life and annual full regeneration. Previous studies revealed that these events are achieved through the proliferation and differentiation of antlerogenic periosteum (AP) cells and pedicle periosteum (PP) cells, respectively. As the cells resident in the AP and the PP possess stem cell attributes, both antler generation and regeneration are stem cell-based processes. However, the cell composition of each tissue type and molecular events underlying antler development remain poorly characterized. Here, we took the approach of single-cell RNA sequencing (scRNA-Seq) and identified eight cell types (mainly THY1+ cells, progenitor cells, and osteochondroblasts) and three core subclusters of the THY1+ cells (SC2, SC3, and SC4). Endothelial and mural cells each are heterogeneous at transcriptional level. It was the proliferation of progenitor, mural, and endothelial cells in the activated antler-lineage-specific tissues that drove the rapid formation of the antler. We detected the differences in the initial differentiation process between antler generation and regeneration using pseudotime trajectory analysis. These may be due to the difference in the degree of stemness of the AP-THY1+ and PP-THY1+ cells. We further found that androgen-RXFP2 axis may be involved in triggering initial antler full regeneration. Fully deciphering the cell composition for these antler tissue types will open up new avenues for elucidating the mechanism underlying antler full renewal in specific and regenerative medicine in general.
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Affiliation(s)
- Hengxing Ba
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Xin Wang
- BGI-Shenzhen, Shenzhen, 518083 Guangdong China
- Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, RNA Institute, Wuhan University, Wuhan, China
| | - Datao Wang
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 130112, Changchun, China
| | - Jing Ren
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Zhen Wang
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Hai-Xi Sun
- BGI-Shenzhen, Shenzhen, 518083 Guangdong China
| | - Pengfei Hu
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Guokun Zhang
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Shengnan Wang
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Chao Ma
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Yusu Wang
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
| | - Enpeng Wang
- Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, 130117 China
| | - Liang Chen
- Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, RNA Institute, Wuhan University, Wuhan, China
| | - Tianbin Liu
- BGI-Shenzhen, Shenzhen, 518083 Guangdong China
- College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049 China
| | - Ying Gu
- BGI-Shenzhen, Shenzhen, 518083 Guangdong China
- College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049 China
- Guangdong Provincial Key Laboratory of Genome Read and Write, BGI-Shenzhen, Shenzhen, 518120 Guangdong China
| | - Chunyi Li
- Institute of Antler Science and Product Technology, Changchun Sci-Tech University, Changchun, 130600 China
- Jilin Provincial Key Laboratory of Deer Antler Biology, Changchun, 130600 China
- College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun, 130118 China
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Zhang W, Ke CH, Guo HH, Xiao L. Antler stem cells and their potential in wound healing and bone regeneration. World J Stem Cells 2021; 13:1049-1057. [PMID: 34567424 PMCID: PMC8422928 DOI: 10.4252/wjsc.v13.i8.1049] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/27/2021] [Revised: 05/10/2021] [Accepted: 07/27/2021] [Indexed: 02/06/2023] Open
Abstract
Compared to other vertebrates, the regenerative capacity of appendages in mammals is very limited. Deer antlers are an exception and can fully regenerate annually in postnatal mammals. This process is initiated by the antler stem cells (AnSCs). AnSCs can be divided into three types: (1) Antlerogenic periosteum cells (for initial pedicle and first antler formation); (2) Pedicle periosteum cells (for annual antler regeneration); and (3) Reserve mesenchyme cells (RMCs) (for rapid antler growth). Previous studies have demonstrated that AnSCs express both classic mesenchymal stem cells (MSCs) and embryonic stem cells (ESCs), and are able to differentiate into multiple cell types in vitro. Thus, AnSCs were defined as MSCs, but with partial ESC attributes. Near-perfect generative wound healing can naturally occur in deer, and wound healing can be achieved by the direct injection of AnSCs or topical application of conditioned medium of AnSCs in rats. In addition, in rabbits, the use of both implants with AnSCs and cell-free preparations derived from AnSCs can stimulate osteogenesis and repair defects of bone. A more comprehensive understanding of AnSCs will lay the foundation for developing an effective clinical therapy for wound healing and bone repair.
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Affiliation(s)
- Wei Zhang
- Medical Imaging Center, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou 510080, Guangdong Province, China
- Research & Development Center, YZ Health-tech Inc., Zhuhai 519000, Guangdong Province, China
| | - Chang-Hong Ke
- Research & Development Center, YZ Health-tech Inc., Zhuhai 519000, Guangdong Province, China
- School of Pharmacy, Jinan University, Guangzhou 510080, Guangdong Province, China
| | - Hai-Hua Guo
- Institute of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, Guangdong Province, China
| | - Li Xiao
- Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou 510080, Guangdong Province, China
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Feleke M, Bennett S, Chen J, Hu X, Williams D, Xu J. New physiological insights into the phenomena of deer antler: A unique model for skeletal tissue regeneration. J Orthop Translat 2020; 27:57-66. [PMID: 33437638 PMCID: PMC7773678 DOI: 10.1016/j.jot.2020.10.012] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/07/2020] [Revised: 10/23/2020] [Accepted: 10/29/2020] [Indexed: 12/14/2022] Open
Abstract
Generally, mammals are unable to regenerate complex tissues and organs however the deer antler provides a rare anomaly to this rule. This osseous cranial appendage which is located on the frontal bone of male deer is capable of stem cell-based organogenesis, annual casting, and cyclic de novo regeneration. A series of recent studies have classified this form of regeneration as epimorphic stem cell based. Antler renewal is initiated by the activation of neural crest derived pedicle periosteal cells (PPCs) found residing within the pedicle periosteum (PP), these PPCs have the potential to differentiate into multiple lineages. Other antler stem cells (ASCs) are the reserve mesenchymal cells (RMCs) located in the antlers tip, which develop into cartilage tissue. Antlerogenic periosteal cells (APCs) found within the antlerogenic periosteum (AP) form the tissues of both the pedicle and first set of antlers. Antler stem cells (ASCs) further appear to progress through various stages of activation, this coordinated transition is considered imperative for stem cell-based mammalian regeneration. The latest developments have shown that the rapid elongation of the main beam and antler branches are a controlled form of tumour growth, regulated by the tumour suppressing genes TP73 and ADAMTS18. Both osteoclastogenesis, as well as osteogenic and chondrogenic differentiation are also involved. While there remains much to uncover this review both summarises and comprehensively evaluates our existing knowledge of tissue regeneration in the deer antler. This will assist in achieving the goal of in vitro organ regeneration in humans by furthering the field of modern regenerative medicine. The Translational potential of this article As a unique stem cell-based organ regeneration process in mammals, the deer antler represents a prime model system for investigating mechanisms of regeneration in mammalian tissues. Novel ASCs could provide cell-based therapies for regenerative medicine and bone remodelling for clinical application. A greater understanding of this process and a more in-depth defining of ASCs will potentiate improved clinical outcomes.
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Affiliation(s)
- Mesalie Feleke
- Division of Regenerative Biology, School of Biomedical Sciences, University of Western Australia, Perth, 6009, Australia
| | - Samuel Bennett
- Division of Regenerative Biology, School of Biomedical Sciences, University of Western Australia, Perth, 6009, Australia
| | - Jiazhi Chen
- Guangdong Provincial Key Laboratory of Industrial Surfactant, Guangdong Research Institute of Petrochemical and Fine Chemical Engineering, Guangdong Academy of Sciences, Guangzhou, 510665, China.,Division of Regenerative Biology, School of Biomedical Sciences, University of Western Australia, Perth, 6009, Australia
| | - Xiaoyong Hu
- Guangdong Provincial Key Laboratory of Industrial Surfactant, Guangdong Research Institute of Petrochemical and Fine Chemical Engineering, Guangdong Academy of Sciences, Guangzhou, 510665, China
| | - Desmond Williams
- Division of Regenerative Biology, School of Biomedical Sciences, University of Western Australia, Perth, 6009, Australia
| | - Jiake Xu
- Division of Regenerative Biology, School of Biomedical Sciences, University of Western Australia, Perth, 6009, Australia
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Wang D, Berg D, Ba H, Sun H, Wang Z, Li C. Deer antler stem cells are a novel type of cells that sustain full regeneration of a mammalian organ-deer antler. Cell Death Dis 2019; 10:443. [PMID: 31165741 PMCID: PMC6549167 DOI: 10.1038/s41419-019-1686-y] [Citation(s) in RCA: 30] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2019] [Revised: 04/19/2019] [Accepted: 05/19/2019] [Indexed: 12/14/2022]
Abstract
Deer antlers are extraordinary mammalian organs that can fully regenerate annually. Antler renewal is a stem cell-based epimorphic process and antler stem (AS) cells can initiate de novo generation of antlers in postnatal mammals. However, although being called stem cells, the AS cells have not been characterized at molecular level based on the stem cell criteria. Comprehensive characterization of the AS cells would undoubtedly help to decipher the mechanism underlying the full regeneration of deer antlers, the only case of stem cell-based epimorphic regeneration in mammals. In the present study, three types of AS cells (antlerogenic periosteal cells APCs, for initial pedicle and first antler formation; pedicle periosteal cells PPC, for annual antler regeneration; and reserve mesenchyme cells RMCs, for rapid antler growth), were isolated for comprehensive molecular characterization. A horn-growth-related gene, RXFP2, was found to be expressed only in AS cells lineages but not in the facial periosteal cells (FPCs, locates geographically in the vicinity of the APCs or PPCs), suggesting the RXFP2 might be a specific marker for the AS cell lineage in deer. Our results demonstrated that AS cells expressed classic MSC markers including surface markers CD73, CD90, CD105 and Stro-1. They also expressed some of the markers including Tert, Nestin, S100A4, nucleostemin and C-Myc, suggesting that they have some attributes of the ESCs. Microinjection of male APC into deer blastocysts resulted in one female foetus (110 days gestation) recovered with obvious pedicle primordia with both male and female genotype detected in the ovary. In conclusion, the AS cells should be defined as MSCs but with partial attributes of ESCs.
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Affiliation(s)
- Datao Wang
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 4899 Juye Street, Changchun, 130112, China
| | - Debbie Berg
- AgResearch Ltd, Ruakura Agricultural Centre, 10 Bisley Road, Hamilton, 3214, New Zealand
| | - Hengxing Ba
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 4899 Juye Street, Changchun, 130112, China
| | - Hongmei Sun
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 4899 Juye Street, Changchun, 130112, China
| | - Zhen Wang
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 4899 Juye Street, Changchun, 130112, China
| | - Chunyi Li
- Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, 4899 Juye Street, Changchun, 130112, China.
- Changchun Sci-Tech University, Changchun, 130600, China.
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Yang F, Wang W, Li J, Haines S, Asher G, Li C. Antler development was inhibited or stimulated by cryosurgery to periosteum or skin in a central antlerogenic region respectively. JOURNAL OF EXPERIMENTAL ZOOLOGY PART B-MOLECULAR AND DEVELOPMENTAL EVOLUTION 2011; 316:359-70. [DOI: 10.1002/jez.b.21409] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/03/2010] [Revised: 01/10/2011] [Accepted: 02/28/2011] [Indexed: 11/08/2022]
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Li C, Suttie JM. Electron microscopic studies of antlerogenic cells from five developmental stages during pedicle and early antler formation in red deer (Cervus elaphus). Anat Rec (Hoboken) 1998; 252:587-99. [PMID: 9845209 DOI: 10.1002/(sici)1097-0185(199812)252:4<587::aid-ar9>3.0.co;2-i] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Previous studies using light microscopy have revealed that histogenesis of deer pedicle and antler has four ossification stages. The first of these stages is the development of the permanent pedicle. Initial development of the pedicle is from the cellular layer cells of the antlerogenic periosteum and these cells have been termed initial antlerogenic cells (IACs). Apart from the IACs, it has also been shown that the cellular layer cells of the apical periosteum/perichondrium, the peripheral periosteum of pedicles or antlers, and the marginal periosteum surrounding the pedicles are also capable of either partially or fully generating a pedicle or an antler. Therefore, these cells can all be considered antlerogenic cells and called apical antlerogenic cells (AACs), peripheral antlerogenic cells (PACs), and marginal antlerogenic cells (MACs), respectively. The aim of this study was to examine the ultrastructure of these antlerogenic cells, and to determine whether there were ultrastructural correlates with the changes of these antlerogenic cells and ossification stages. The ultrastructure of each type of antlerogenic cells was systematically examined using transmission electron microscopy, at each stage of pedicle and first antler growth. At the first ossification stage, the IACs were spindle-shaped and inactive. The most obvious feature was the presence of abundant intracellular glycogen. The MACs were similar to the IACs. During the early second stage, most of the AACs changed in appearance from preosteoblasts to prechondroblasts. Much less heterochromatin was found in the AACs than in the IACs. The most striking attribute of the AACs was the existence of intracellular collagen fibers. The MACs showed abnormal dilation of the rough endoplasmic reticulum (RER). During the late second stage, the majority of the AACs were prechondroblasts. AAC nucleoli were clearly discernible and the cisternae of the RER were arranged in parallel. The MACs contained a greater proportion of abnormally-dilated RER. During the third stage, the AACs were all prechondroblasts. The Golgi apparatus in these cells was well developed. Many free ribosomes in rosettes were scattered in the cytoplasm. Most cytoplasm of the majority of the MACs was occupied by abnormally-dilated RER (the lumen of the RER was extremely dilated and appeared electron-lucent). During the fourth stage, the AACs were similar to their counterparts from the third stage, but the boundaries of some AACs were ill-defined. Some MACs were found to be undergoing apoptosis. The PACs were becoming less and less active from distal to proximal along the shaft of the antler. It is a novel finding that antlerogenic cells change in appearance and subcellular content from preosteoblasts to prechondroblasts prior to the transition from intramembranous to endochondral ossification during pedicle formation. Therefore, the differentiation process from antlerogenic cells to chondroblasts is a matter of maturation from prechondroblasts to chondroblasts. The fact that the antlerogenic cells are rich in glycogen makes them more like embryonic cells. The local membrane deficiency of some AACs at the fourth stage and the presence of mature collagen fibrils within the AACs may reflect the unusually high demand for collagen fibrils during the period of rapid antler growth.
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Affiliation(s)
- C Li
- AgResearch, Invermay Agricultural Centre, Mosgiel, New Zealand
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Li C, Waldrup KA, Corson ID, Littlejohn RP, Suttie JM. Histogenesis of antlerogenic tissues cultivated in diffusion chambers in vivo in red deer (Cervus elaphus). THE JOURNAL OF EXPERIMENTAL ZOOLOGY 1995; 272:345-55. [PMID: 7673872 DOI: 10.1002/jez.1402720504] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
In a previous study we showed that formation of deer pedicle and first antler proceeded through four ossification pattern change stages: intramembranous, transition, pedicle endochondral, and antler endochondral. In the present study antlerogenic tissues (antlerogenic periosteum, apical periosteum/perichondrium, and apical perichondrial of pedicle and antler) taken from four developmental stages were cultivated in diffusion chambers in vivo as autografts for 42-68 days. The results showed that all the cultivated tissues without exception formed trabecular bone de novo, irrespective of whether they were forming osseous, osseocartilaginous, or cartilaginous tissue at the time of initial implant surgery; in two cases in the apical perichondria from antler group, avascularized cartilage also formed. Therefore, the antlerogenic cells, like the progenitor cells of somatic secondary type cartilage, have a tendency to differentiate into osteoblasts and then form trabecular bone. Consequently, the differentiation pathway whereby antlerogenic cells change from forming osteoblasts to forming chondroblasts during pedicle formation is caused by extrinsic factors. Both oxygen tension and mechanical pressure are postulated to be the factors that cause this alteration of the differentiation pathway.
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Affiliation(s)
- C Li
- AgResearch, Invermay Agricultural Centre, Mosgiel, New Zealand
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Abstract
Through a series of interrogatories, unsolved problems of antler evolution, anatomy, development, physiology, and pathology are probed, with commentaries, on the following prospects for future research: 1. How could these improbable appendages have evolved mechanisms to commit suicide, jettison the corpse, and regenerate new ones every year? 2. By what developmental processes are antlers able to prescribe their own morphogenesis with mirror image accuracy year after year and in some cases produce deliberate asymmetries? 3. What causes the scalp to transform into velvet skin as a deer's first antlers develop? 4. Why do healing pedicle stumps give rise to antler buds instead of scar tissue? 5. How is the unprecedented rate of antler elongation related to the diameter and length of the structure to be grown? 6. How come wound healing by pedicle skin is held in abeyance for several months until new growth resumes? 7. How is it that tropical deer regenerate antlers at any time of year, while in temperate zones deer do so in seasonal unison? 8. How do deer find enough calcium to make such massive antlers in only a few months? 9. What is the nature of the bizarre tumors that some antlers grow following castration?
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Affiliation(s)
- R J Goss
- Division of Biology and Medicine, Brown University, Providence, Rhode Island 02912, USA
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