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Global analysis of tissue-differential gene expression patterns and functional regulation of rapid antler growth

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Abstract

Deer antlers are unique mammalian appendages that grow faster than any other known organ. During the rapid growth stage at 60 days after casting the previous antler, the maximal antler growth rate is up to or even more than 2 cm/day. Antler growth is driven by the growth center located in the antler tip. The growth center consists of several tissue layers from distal to proximal, including the skin, mesenchyme, precartilage, and cartilage. To analyze the gene expression patterns of the antler growth center in a tissue-differential manner and explore the molecular mechanism responsible for rapid antler growth, we used an RNA-Seq method to analyze gene expression patterns in different tissues of the Sika deer antler growth center during the rapid growth stage. We demonstrated considerable diversity in the expression levels of functional genes among different tissues within the antler growth center. These tissue-differentially expressed genes included transcription factors, growth factors, and extracellular matrix proteins. We identified a series of genes that contribute to chondrogenesis from mesenchymal cell condensation to chondrocyte differentiation. The genes identified in the antler growth center at the rapid growth stage provide valuable insight into the physiological mechanisms underlying rapid antler growth.

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References

  • Akiyama H, Chaboissier MC, Martin JF, Schedl A, de Crombrugghe B (2002) The transcription factor Sox9 has essential roles in successive steps of the chondrocyte differentiation pathway and is required for expression of Sox5 and Sox6. Genes Dev 16:2813–2828

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arnott JA, Lambi AG, Mundy C, Hendesi H, Pixley RA, Owen TA, Safadi FF, Popoff SN (2011) The role of connective tissue growth factor (CTGF/CCN2) in skeletogenesis. Crit Rev Eukaryot Gene Expr 21:43–69

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bhattaram P, Penzo-Méndez A, Kato K, Bandyopadhyay K, Gadi A, Taketo MM, Lefebvre V (2014) SOXC proteins amplify canonical WNT signaling to secure nonchondrocytic fates in skeletogenesis. J Cell Biol 207:657–671

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bouderlique T, Henault E, Lebouvier A, Frescaline G, Bierling P, Rouard H, Courty J, Albanese P, Chevallier N (2014) Pleiotrophin commits human bone marrow mesenchymal stromal cells towards hypertrophy during chondrogenesis. PLoS One 9:e88287

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen H, Ghori-Javed FY, Rashid H, Adhami MD, Serra R, Gutierrez SE, Javed A (2014) Runx2 regulates endochondral ossification through control of chondrocyte proliferation and differentiation. J Bone Miner Res 29:2653–2665

    Article  CAS  PubMed  Google Scholar 

  • Chijimatsu R, Kunugiza Y, Taniyama Y, Nakamura N, Tomita T, Yoshikawa H (2015) Expression and pathological effects of periostin in human osteoarthritis cartilage. BMC Musculoskelet Disord 16:215

    Article  PubMed  PubMed Central  Google Scholar 

  • Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676

    Article  CAS  PubMed  Google Scholar 

  • Cserjesi P, Brown D, Ligon KL, Lyons GE, Copeland NG, Gilbert DJ (1995) Scleraxis: a basic helix-loop-helix protein that prefigures skeletal formation during mouse embryogenesis. Development 121pp:1099–1110

    Google Scholar 

  • D’Angelo M, Yan Z, Nooreyazdan M, Pacifici M, Sarment DS, Billings PC, Leboy PS (2000) MMP-13 is induced during chondrocyte hypertrophy. J Cell Biochem 77:678–693

    Article  PubMed  Google Scholar 

  • Darby IA, Bisucci T, Raghoenath S, Olsson J, Muscat GE, Koopman P (2001) Sox18 is transiently expressed during angiogenesis in granulation tissue of skin wounds with an identical expression pattern to Flk-1 mRNA. Lab Investig 81:937–943

    Article  CAS  PubMed  Google Scholar 

  • Eckert RL, Adhikary G, Young CA, Jans R, Crish JF, Xu W, Rorke EA (2013) AP1 transcription factors in epidermal differentiation and skin cancer. J Skin Cancer 2013:537028

    Article  PubMed  PubMed Central  Google Scholar 

  • Erdag G, Medalie DA, Rakhorst H, Krueger GG, Morgan JR (2004) FGF-7 expression enhances the performance of bioengineered skin. Mol Ther 10:76–85

    Article  CAS  PubMed  Google Scholar 

  • Gallitzendoerfer R, Abouzied MM, Hartmann D, Dobrowolski R, Gieselmann V, Franken S (2008) Hepatoma-derived growth factor (HDGF) is dispensable for normal mouse development. Dev Dyn 237:1875–1885

    Article  CAS  PubMed  Google Scholar 

  • Green JD, Tollemar V, Dougherty M, Yan Z, Yin L, Ye J, Collier Z, Mohammed MK, Haydon RC, Luu HH, Kang R, Lee MJ, Ho SH, He TC, Shi LL, Athiviraham A (2015) Multifaceted signaling regulators of chondrogenesis: implications in cartilage regeneration and tissue engineering. Genes Dis 2:307–327

    Article  PubMed  PubMed Central  Google Scholar 

  • Gu L, Mo E, Zhu X, Jia X, Fang Z, Sun B, Sung C (2008) Analysis of gene expression in four parts of the red-deer antler using DNA chip microarray technology. Anim Biol 58:67–90

    Article  Google Scholar 

  • Guha S, Goyal SP, Kashyap VK (2007) Molecular phylogeny of musk deer: a genomic view with mitochondrial 16S rRNA and cytochrome b gene. Mol Phylogenet Evol 42:585–597

    Article  CAS  PubMed  Google Scholar 

  • Gyurján I Jr, Molnár A, Borsy A, Stéger V, Hackler L Jr, Zomborszky Z, Papp P, Duda E, Deák F, Lakatos P, Puskás LG, Orosz L (2007) Gene expression dynamics in deer antler: mesenchymal differentiation toward chondrogenesis. Mol Gen Genomics 277:221–235

    Article  CAS  Google Scholar 

  • Hassanin A, Douzery EJ (2005) Molecular and morphological phylogenies of ruminantia and the alternative position of the moschidae. Syst Biol 52:206–228

    Article  Google Scholar 

  • Hellman M, Arumäe U, Yu LY, Lindholm P, Peränen J, Saarma M, Permi P (2011) Mesencephalic astrocyte-derived neurotrophic factor (MANF) has a unique mechanism to rescue apoptotic neurons. J Biol Chem 286:2675–2680

    Article  CAS  PubMed  Google Scholar 

  • Hrdlickova R, Toloue M, Tian B (2017) RNA-Seq methods for transcriptome analysis. Wiley Interdiscip Rev RNA 8:1364

    Article  CAS  Google Scholar 

  • Ivkovic S, Yoon BS, Popoff SN, Safadi FF, Libuda DE, Stephenson RC, Daluiski A, Lyons KM (2003) Connective tissue growth factor coordinates chondrogenesis and angiogenesis during skeletal development. Development 130:2779–2791

    Article  CAS  PubMed  Google Scholar 

  • Kaback LA, do Soung Y, Naik A, Smith N, Schwarz EM, O'Keefe RJ, Drissi H (2008) Osterix/Sp7 regulates mesenchymal stem cell mediated endochondral ossification. J Cell Physiol 214:173–182

    Article  CAS  PubMed  Google Scholar 

  • Kafienah W, Cheung FL, Sims T, Martin I, Miot S, Von Ruhland C, Roughley PJ, Hollander AP (2008) Lumican inhibits collagen deposition in tissue engineered cartilage. Matrix Biol 27:526–534

    Article  CAS  PubMed  Google Scholar 

  • Kameda T, Watanabe H, Iba H (1997) C-Jun and JunD suppress maturation of chondrocytes. Cell Growth Differ 8:495–503

    CAS  PubMed  Google Scholar 

  • Kato K, Bhattaram P, Penzo-Méndez A, Gadi A, Lefebvre V (2015) SOXC transcription factors induce cartilage growth plate formation in mouse embryos by promoting noncanonical WNT signaling. J Bone Miner Res 30:1560–1571

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi T, Chung UI, Schipani E, Starbuck M, Karsenty G, Katagiri T, Goad DL, Lanske B, Kronenberg HM (2002) PTHrP and Indian hedgehog control differentiation of growth plate chondrocytes at multiple steps. Development 129:2977–2986

    CAS  PubMed  Google Scholar 

  • Li C, Clark DE, Lord EA, Stanton JA, Suttie JM (2002) Sampling technique to discriminate the different tissue layers of growing antler tips for gene discovery. Anat Rec 268:125–130

    Article  CAS  PubMed  Google Scholar 

  • Lin SY, Yang J, Everett AD, Clevenger CV, Koneru M, Mishra PJ, Kamen B, Banerjee D, Glod J (2008) The isolation of novel mesenchymal stromal cell chemotactic factors from the conditioned medium of tumor cells. Exp Cell Res 314:3107–3117

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu M, Yao B, Zhang H, Guo H, Hu D, Wang Q, Zhao Y (2014) Identification of novel reference genes using sika deer antler transcriptome expression data and their validation for quantitative gene expression analysis. Genes & Genomics 36:573–582

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C (T)) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Marie PJ, Haÿ E, Modrowski D, Revollo L, Mbalaviele G, Civitelli R (2014) Cadherin-mediated cell-cell adhesion and signaling in the skeleton. Calcif Tissue Int 94:46–54

    Article  CAS  PubMed  Google Scholar 

  • Masago Y, Hosoya A, Kawasaki K, Kawano S, Nasu A, Toguchida J, Fujita K, Nakamura H, Kondoh G, Nagata K (2012) The molecular chaperone Hsp47 is essential for cartilage and endochondral bone formation. J Cell Sci 125:1118–1128

    Article  CAS  PubMed  Google Scholar 

  • Michalak M, Groenendyk J, Szabo E, Gold LI, Opas M (2009) Calreticulin, a multi-process calcium-buffering chaperone of the endoplasmic reticulum. Biochem J 417:651–666

    Article  CAS  PubMed  Google Scholar 

  • Miljkovic ND, Cooper GM, Marra KG (2008) Chondrogenesis, bone morphogenetic protein-4 and mesenchymal stem cells. Osteoarthr Cartil 16:112111–112130

    Article  Google Scholar 

  • Molnár A, Gyurján I, Korpos E, Borsy A, Stéger V, Buzás Z, Kiss I, Zomborszky Z, Papp P, Deák F, Orosz L (2007) Identification of differentially expressed genes in the develo** antler of red deer Cervus elaphus. Mol Gen Genomics 277:237–248

    Article  CAS  Google Scholar 

  • Morello R, Bertin TK, Schlaubitz S, Shaw CA, Kakuru S, Munivez E, Hermanns P, Chen Y, Zabel B, Lee B (2008) Brachy-syndactyly caused by loss of Sfrp2 function. J Cell Physiol 217:127–137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Murphy LI, Fischer D, Chiquet-Ehrismann R, Mackie EJ (2000) Tenascin-C induced stimulation of chondrogenesis is dependent on the presence of the C-terminal fibrinogen-like globular domain. FEBS Lett 480:189–192

    Article  CAS  PubMed  Google Scholar 

  • Nakamura Y, Yamamoto K, He X, Otsuki B, Kim Y, Murao H, Soeda T, Tsumaki N, Deng JM, Zhang Z, Behringer RR, Bd C, Postlethwait JH, Warman ML, Nakamura T, Akiyama H (2011) Wwp2 is essential for palatogenesis mediated by the interaction between Sox9 and mediator subunit 25. Nat Commun 2:251

    Article  CAS  PubMed  Google Scholar 

  • Ogata A, Shimizu T, Abe R, Shimizu H, Sakai M (2004) Expression of c-maf and mafB genes in the skin during rat embryonic development. Acta Histochem 106:65–67

    Article  CAS  PubMed  Google Scholar 

  • Park HJ, Lee DH, Park SG, Lee SC, Cho S, Kim HK, Kim JJ, Bae H, Park BC (2004) Proteome analysis of red deer antlers. Proteomics 4:3642–3653

    Article  CAS  PubMed  Google Scholar 

  • Patel HB, Kornerup KN, Sampaio AL, D'Acquisto F, Seed MP, Girol AP, Gray M, Pitzalis C, Oliani SM, Perretti M (2012) The impact of endogenous annexin A1 on glucocorticoid control of inflammatory arthritis. Ann Rheum Dis 71:1872–1880

    Article  CAS  PubMed  Google Scholar 

  • Pita-Thomas W, Fernández-Martos C, Yunta M, Maza RM, Navarro-Ruiz R, Lopez-Rodríguez MJ (2010) Gene expression of axon growth promoting factors in the deer antler. PLoS One 5:e15706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Price J, Allen S (2004) Exploring the mechanisms regulating regeneration of deer antlers. Philos Trans R Soc Lond Ser B Biol Sci 359:809–822

    Article  CAS  Google Scholar 

  • Reppe S, Rian E, Jemtland R, Olstad OK, Gautvik VT, Gautvik KM (2000) Sox-4 messenger RNA is expressed in the embryonic growth plate and regulated via the parathyroid hormone/parathyroid hormone-related protein receptor in osteoblast-like cells. J Bone Miner Res 15:2402–2412

    Article  CAS  PubMed  Google Scholar 

  • Ricard-Blum S (2011) The collagen family. Cold Spring Harb Perspect Biol 3:a004978

    Article  PubMed  PubMed Central  Google Scholar 

  • Rollman O, Jensen UB, Ostman A, Bolund L, Gústafsdóttir SM, Jensen TG (2003) Platelet derived growth factor (PDGF) responsive epidermis formed from human keratinocytes transduced with the PDGF beta receptor gene. J Invest Dermatol 120:742–749

    Article  CAS  PubMed  Google Scholar 

  • Rotllant J, Liu D, Yan YL, Postlethwait JH, Westerfield M, Du SJ (2008) Sparc (Osteonectin) functions in morphogenesis of the pharyngeal skeleton and inner ear. Matrix Biol 27:561–572

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schmid R, Meyer K, Spang R, Schittek B, Bosserhoff AK (2013) YBX1 is a modulator of MIA/CD-RAP-dependent chondrogenesis. PLoS One 8:e82166

    Article  PubMed  PubMed Central  Google Scholar 

  • Suemoto H, Muragaki Y, Nishioka K, Sato M, Ooshima A, Itoh S, Hatamura I, Ozaki M, Braun A, Gustafsson E, Fässler R (2007) Trps1 regulates proliferation and apoptosis of chondrocytes through Stat3 signaling. Dev Biol 312:572–581

    Article  CAS  PubMed  Google Scholar 

  • Tay LX, Lim CK, Mansor A, Kamarul T (2014) Differential protein expression between chondrogenic differentiated MSCs, undifferentiated MSCs and adult chondrocytes derived from Oryctolagus cuniculus in vitro. Int J Med Sci 11:24–33

    Article  CAS  PubMed  Google Scholar 

  • Tekari A, Luginbuehl R, Hofstetter W, Egli RJ (2015) Transforming growth factor beta signaling is essential for the autonomous formation of cartilage-like tissue by expanded chondrocytes. PLoS One 10:e0120857

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Toulza E, Mattiuzzo NR, Galliano MF, Jonca N, Dossat C, Jacob D, de Daruvar A, Wincker P, Serre G, Guerrin M (2007) Large-scale identification of human genes implicated in epidermal barrier function. Genome Biol 8:R107

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L (2010) Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28:511–515

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Truzzi F, Marconi A, Pincelli C (2011) Neurotrophins in healthy and diseased skin. Dermatoendocrinol 3:32–36

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Twomey JD, Thakore P, Hartman DA, Myers EG, Hsieh AH (2014) Roles of type VI collagen and decorin in human mesenchymal stem cell biophysics during chondrogenic differentiation. Eur Cell Mater 27:237–250

    Article  CAS  PubMed  Google Scholar 

  • Wang W, Lian N, Li L, Moss HE, Wang W, Perrien DS, Elefteriou F, Yang X (2009) Atf4 regulates chondrocyte proliferation and differentiation during endochondral ossification by activating Ihh transcription. Development 136:4143–4153

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wuelling M, Kaiser FJ, Buelens LA, Braunholz D, Shivdasani RA, Dep** R, Vortkamp A (2009) Trps1, a regulator of chondrocyte proliferation and differentiation, interacts with the activator form of Gli3. Dev Biol 328:40–53

    Article  CAS  PubMed  Google Scholar 

  • **e C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li CY, Wei L (2011) KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res 39:W316–W322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yao B, Zhang M, Liu M, Wang Q, Liu M, Zhao Y (2018) Sox9 functions as a master regulator of antler growth by controlling multiple cell lineages. DNA Cell Biol 37:15–22

    Article  CAS  PubMed  Google Scholar 

  • Yao B, Zhao Y, Wang Q, Zhang M, Liu M, Liu H, Li J (2012a) De novo characterization of the antler tip of Chinese Sika deer transcriptome and analysis of gene expression related to rapid growth. Mol Cell Biochem 364:93–100

    Article  CAS  PubMed  Google Scholar 

  • Yao B, Zhao Y, Zhang H, Zhang M, Liu M, Liu H, Li J (2012b) Sequencing and de novo analysis of the Chinese Sika deer antler-tip transcriptome during the ossification stage using Illumina RNA-Seq technology. Biotechnol Lett 34:813–822

    Article  CAS  PubMed  Google Scholar 

  • Youssef A, Aboalola D, Han VK (2017) The roles of insulin-like growth factors in mesenchymal stem cell niche. Stem Cells Int 2017:9453108

    PubMed  PubMed Central  Google Scholar 

  • Zeisberg M, Neilson EG (2009) Biomarkers for epithelial-mesenchymal transitions. J Clin Invest 119:1429–1437

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang JY, Green CL, Tao S, Khavari PA (2004) NF-kappaB RelA opposes epidermal proliferation driven by TNFR1 and JNK. Genes Dev 18:17–22

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This work was financially supported through grants from the Science and Technology Development Project of Jilin Province (Grant number 20170520044JH), the Science and Technology Project of Jilin Provincial Education Department (Grant number JJKH20170721KJ), the TCM Clinical Research Center for Bone diseases of Jilin Province (Grant number 20180623048TC), and the National Natural Science Foundation of China (Grant number 81702136).

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Authors and Affiliations

  1. Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, 130117, China

    Bao** Yao, Daqing Zhao & Yu Zhao

  2. Innovation Practice Center, Changchun University of Chinese Medicine, Changchun, 130117, China

    Mei Zhang

  3. The Affiliated Hospital of Changchun University of Chinese Medicine, Changchun, 130117, China

    Hongwei Gao, Bocheng Lu & **angyang Leng

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  1. Bao** Yao
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  2. Mei Zhang
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  5. **angyang Leng
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  6. Daqing Zhao
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  7. Yu Zhao
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Correspondence to Daqing Zhao or Yu Zhao.

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Communicated by: Shuiqiao Yuan

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Fig. S1

Histological structure of antler tips (Li et al. 2002). a Antler ready to be harvested, and line indicating amount of tip removed. b Tip after being cut sagittally. c Layers as identified by the distinct morphological markers and marked by the stitches. d BrdU incorporation in the dermis (D), outer reserve mesenchyme (ORM), and inner reserve mesenchyme (IRM). e Histological section of the antler tip with holes from the stitches evident (arrows). D, dermis; RM, reservemesenchyme; PC, precartilage; TZ, transition zone; C, cartilage. (PNG 1259 kb)

High Resolution Image (TIF 1643 kb)

Fig. S2

Histogram of GO classification of assembled sequences. The results were grouped into three main categories: biological process, cellular component, and molecular function. The ordinate is the name of the GO term, while the abscissa is the number of genes belonging to this GO term. (PNG 996 kb)

High-resolution image (TIF 1141 kb)

Fig. S3

Histogram of KEGG pathway annotation of assembled sequences. The results were grouped into six main categories: cellular processes, environmental information processing, genetic information processing, metabolism, and organismal systems. The ordinate is the name of the pathway, while the abscissa is the number of genes belonging to this pathway. (PNG 2775 kb)

High-resolution image (TIF 5036 kb)

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Yao, B., Zhang, M., Gao, H. et al. Global analysis of tissue-differential gene expression patterns and functional regulation of rapid antler growth. Mamm Res 64, 235–248 (2019). https://doi.org/10.1007/s13364-018-0394-9

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