FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / BMC Res Notes / Decoding the transcriptomic expression and genomic methylati...
BMC research notes2023; 16(1); 267; doi: 10.1186/s13104-023-06562-1

Decoding the transcriptomic expression and genomic methylation patterns in the tendon proper and its peritenon region in the aging horse.

Abstract: Equine tendinopathies are challenging because of the poor healing capacity of tendons commonly resulting in high re-injury rates. Within the tendon, different regions - tendon proper (TP) and peritenon (PERI) - contribute to the tendon matrix in differing capacities during injury and aging. Aged tendons have decreased repair potential; the underlying transcriptional and epigenetic changes that occur in the TP and PERI regions are not well understood. The objective of this study was to assess TP and PERI regional differences in adolescent, midlife, and geriatric horses using RNA sequencing and DNA methylation techniques. Results: Differences existed between TP and PERI regions of equine superficial digital flexor tendons by age as evidenced by RNASeq and DNA methylation. Cluster analysis indicated that regional distinctions existed regardless of age. Genes such as DCN, COMP, FN1, and LOX maintained elevated TP expression while genes such as GSN and AHNAK were abundant in PERI. Increased gene activity was present in adolescent and geriatric populations but decreased during midlife. Regional differences in DNA methylation were also noted. Notably, when evaluating all ages of TP against PERI, five genes (HAND2, CHD9, RASL11B, ADGRD1, and COL14A1) had regions of differential methylation as well as differential gene expression.
© 2023. BioMed Central Ltd., part of Springer Nature.
Get Full Text
Publication Date: 2023-10-11 PubMed ID: 37821884PubMed Central: PMC10566085DOI: 10.1186/s13104-023-06562-1Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research is focused on understanding the changes in transcriptomic expression and genomic methylation in different parts of the tendon in horses as they age. This is achieved through the comparison of these patterns in the two regions of tendons known as the tendon proper and peritenon in horses during different stages of life – adolescence, middle age, and old age.

Study Overview and Objective

  • The primary goal of this research was to investigate the differences in the regions of the tendon proper (TP) and peritenon (PERI) in horses during different life stages. This was achieved by utilizing RNA sequencing and DNA methylation techniques.
  • The researchers wanted to understand the varying contributions of these two regions to the tendon matrix during aging and injury. The differences in transcriptional and epigenetic changes in these areas are not well understood, especially when considering the decreased repair potential of aged tendons.

Results of the Research

  • The research results revealed significant differences in the TP and PERI regions of equine superficial digital flexor tendons based on age. This was determined using the RNASeq results and DNA methylation patterns.
  • The cluster analysis also revealed that there are regional differences that exist beyond the normal impacts of age.
  • For instance, some genes like DCN, COMP, FN1, and LOX were found to have elevated expression in TP while genes like GSN and AHNAK were more abundant in PERI.

Implications of the Findings

  • The results showed that there are regional variations in gene activity in adolescent and geriatric populations, with a dip in activity during midlife.
  • The DNA methylation patterns also showed regional differences.
  • Significantly, the study discovered that there are five genes (HAND2, CHD9, RASL11B, ADGRD1, and COL14A1) which show differences in both methylation and gene expression when comparing all ages of TP against PERI. This discovery could have implications for our understanding of tendon degradation and problems in aging horses.”

Conclusion

  • Overall, this research provides new insights into the genomic and epigenetic changes in TP and PERI regions of equine tendons across different age groups, and opens up avenues for further investigation into the impact of these changes on the healing capacity and injury rates of aged horses’ tendons.

Cite This Article

APA
Pechanec MY, Mienaltowski MJ. (2023). Decoding the transcriptomic expression and genomic methylation patterns in the tendon proper and its peritenon region in the aging horse. BMC Res Notes, 16(1), 267. https://doi.org/10.1186/s13104-023-06562-1

Publication

BMC research notes
ISSN: 1756-0500
NlmUniqueID: 101462768
Country: England
Language: English
Volume: 16
Issue: 1
Pages: 267
PII: 267

Researcher Affiliations

Pechanec, Monica Y
  • Department of Animal Science, University of California Davis, 2251 Meyer Hall, One Shields Ave, Davis, CA, 95616, USA.
Mienaltowski, Michael J
  • Department of Animal Science, University of California Davis, 2251 Meyer Hall, One Shields Ave, Davis, CA, 95616, USA. mjmienaltowski@ucdavis.edu.

MeSH Terms

  • Horses / genetics
  • Animals
  • Transcriptome
  • Methylation
  • Tendons / metabolism
  • Aging / genetics
  • Genomics
  • Horse Diseases

Grant Funding

  • D17EQ-818 / Morris Animal Foundation, United States

Conflict of Interest Statement

The authors declare no competing interests.

References

This article includes 58 references
  1. Tipton TE, Ray CS, Hand DR. Superficial digital flexor tendonitis in cutting horses: 19 cases (2007–2011). J Am Vet Med A 2013;243:1162–5.
    doi: 10.2460/javma.243.8.1162pubmed: 24094264google scholar: lookup
  2. Thorpe CT, Glegg PD, Birch HL. A review of tendon injury: why is the equine superficial digital flexor tendon most at risk?. Equine Vet J 2010;42(2):174–80.
    doi: 10.2746/042516409X480395pubmed: 20156256google scholar: lookup
  3. Dyson SJ. Medical management of superficial digital flexor tendonitis: a comparative study in 219 horses (1992–2000). Equine Vet J 2004;36:415–9.
    doi: 10.2746/0425164044868422pubmed: 15253082google scholar: lookup
  4. Thorpe CT, Stark RJF, Goodship AE, Birch HL. Mechanical properties of the equine superficial digital flexor tendon relate to specific collagen cross-link levels. Equine Vet J Suppl 2010;38:538–43.
    doi: 10.1111/j.2042-3306.2010.00175.xpubmed: 21059057google scholar: lookup
  5. Murray RC, Dyson SJ, Tranquille C, Adams V. Association of type of sport and performance level with anatomical site of orthopaedic injury diagnosis. Equine Vet J Suppl 2006; (36):411–6.
    pubmed: 17402457
  6. Sugiyama Y, Naito K, Goto K, Kojima Y, Furuhata A, Igarashi M, Nagaoka I, Kaneko K. Effect of aging on the tendon structure and tendon–associated gene expression in mouse foot flexor tendon. Biomedical Rep 2019;10:238–44.
    pmc: PMC6439433pubmed: 30972219
  7. Snedeker JG, Foolen J. Tendon injury and repair – a perspective on the basic mechanisms of tendon Disease and future clinical therapy. Acta Biomat 2017;63:18–36.
    doi: 10.1016/j.actbio.2017.08.032pubmed: 28867648google scholar: lookup
  8. Avery NC, Bailey AJ. Enzymatic and non-enzymatic cross-linking mechanisms in relation to turnover of collagen: relevance to aging and exercise. Scan J Med Sci Sports 2005;15(4):231–40.
    doi: 10.1111/j.1600-0838.2005.00464.xpubmed: 15998340google scholar: lookup
  9. Ackerman JE, Bah I, Jonason JH, Buckley MR, Loiselle AE. Aging does not alter tendon mechanical properties during homeostasis, but does impair flexor tendon healing. J Orthop Res 2017;35(12):2716–24.
    doi: 10.1002/jor.23580pmc: PMC5645212pubmed: 28419543google scholar: lookup
  10. Dunkman AA, Buckley MR, Mienaltowski MJ, Adams SM, Thomas SJ, Satchell L, Kumar A, Pathmanathan L, Beason DP, Iozzo RV, Birk DE, Soslowsky LJ. Decorin expression is important for age-related changes in tendon structure and mechanical properties. Matrix Biol 2013;32:3–13.
    doi: 10.1016/j.matbio.2012.11.005pmc: PMC3615887pubmed: 23178232google scholar: lookup
  11. Andarawis-Puri N, Flatow EL, Soslowsky LJ. Tendon basic science: development, repair, regeneration, and healing. J Orthop Res 2015;33:780–4.
    doi: 10.1002/jor.22869pmc: PMC4427041pubmed: 25764524google scholar: lookup
  12. Mienaltowski MJ, Dunkman AA, Buckley MR, Beason DP, Adams SM, Birk DE, Soslowsky LJ. Injury response of geriatric mouse patellar tendons. J Orthop Res 2016;34:1256–63.
    doi: 10.1002/jor.23144pmc: PMC4919222pubmed: 26704368google scholar: lookup
  13. Wu YT, Wu PT, Jou IM. Peritendinous elastase treatment induces tendon degeneration in rats: a potential model of tendinopathy in vivo. J Orthop Res 2016;34:471–7.
    doi: 10.1002/jor.23030pubmed: 26291184google scholar: lookup
  14. Kostrominova TY, Brooks SV. Age-related changes in structure and extracellular matrix protein expression levels in rat tendons. Age (Dordr) 2013;35:2203–14.
    doi: 10.1007/s11357-013-9514-2pmc: PMC3824999pubmed: 23354684google scholar: lookup
  15. Thankam FG, Boosani CS, Dilisio MF, Agrawal DK. Epigenetic mechanisms and implications in tendon inflammation (review). Int J Mol Med 2019;43(1):3–14.
    pmc: PMC6257858pubmed: 30387824
  16. Riasat K, Bardell D, Goljanek-Whysall K, Clegg PD, Peffers MJ. Epigenetic mechanisms in Tendon Ageing. Br Med Bull 2020; 1–18.
    pmc: PMC7585832pubmed: 32827252
  17. Mienaltowski MJ, Adams SM, Birk DE. Regional differences in stem cell/progenitor cell populations from the mouse Achilles tendon. Tissue Eng Part A 2013;19:199–210.
    doi: 10.1089/ten.tea.2012.0182pmc: PMC3530943pubmed: 22871316google scholar: lookup
  18. Mienaltowski MJ, Adams SM, Birk DE. Tendon proper- and peritenon- derived progenitor cells have unique tenogenic properties. Stem Cell Res Ther 2014;5(4):86.
    doi: 10.1186/scrt475pmc: PMC4230637pubmed: 25005797google scholar: lookup
  19. Dyment NA, Liu CF, Kazemi N, Aschbacher-Smith LE, Kenter K, Breidenbach AP, Shearn JT, Wylie C, Rowe DW, Butler DL. The paratenon contributes to scleraxis-expressing cells during patellar tendon healing. PLoS ONE 2013;8(3):e59944.
    doi: 10.1371/journal.pone.0059944pmc: PMC3608582pubmed: 23555841google scholar: lookup
  20. Kaji DA, Howell KL, Balic Z, Hubmacher D, Huang A. Tgfb signaling is required for tenocyte recruitment and functional neonatal tendon regeneration. eLife 2020;9:e51779.
    doi: 10.7554/eLife.51779pmc: PMC7324157pubmed: 32501213google scholar: lookup
  21. Doherty R, Couldrey C. Exploring genome wide bisulfite sequencing for DNA methylation analysis in livestock: a technical assessment. Front Genet 2014;5:126.
    doi: 10.3389/fgene.2014.00126pmc: PMC4026711pubmed: 24860595google scholar: lookup
  22. Hubert M, Rousseeuw PJ, Vanden Branden K. ROBPCA: a new approach to robust principal components analysis. Technometrics 2005;47:64–79.
    doi: 10.1198/004017004000000563google scholar: lookup
  23. Todorov V, Filzmoser P. An object oriented Framework for Robust Multivariate Analysis. J Stat Soft 2009;32(3):1–47.
    doi: 10.18637/jss.v032.i03google scholar: lookup
  24. Jelinsky SA, Rodeo SA, Li J, Gulotta LV, Archambault JM, SeeHerman HJ. Regulation of gene expression in human tendinopathy. BMC Musculoskelet Disord 2011;12:86.
    doi: 10.1186/1471-2474-12-86pmc: PMC3095578pubmed: 21539748google scholar: lookup
  25. Feldt J, Schicht M, Garreis F, Welss J, Schneider UW, Paulsen F. Structure, regulation and related Diseases of the actin-binding protein gelsolin. Expert Rev Mol Med 2019;20:e7.
    doi: 10.1017/erm.2018.7pubmed: 30698126google scholar: lookup
  26. Cadby JA, Buehler E, Godbout C, van Weeren PR, Snedeker JG. Different between the cell populations from the Peritenon and Tendon Core with Regard to their potential implication in Tendon Repair. PLoS ONE 2014;9(3):e92474.
    doi: 10.1371/journal.pone.0092474pmc: PMC3961373pubmed: 24651449google scholar: lookup
  27. Howell K, Chien C, Bell R, Laudier D, Trufa SF, Keene DR, Andarawis-Puri N, Huang A. Novel model of Tendon Regeneration reveals distinct cell mechanisms underlying regenerative and fibrotic Tendon Healing. Sci Rep 2017;7:45238.
    doi: 10.1038/srep45238pmc: PMC5362908pubmed: 28332620google scholar: lookup
  28. Xu W, Sun Y, Zhang J, Xu K, Pan L, He L, Song Y, Njunge L, Xu Z, Chiang MYM, Sung KLP, Chuong CM, Yang Li. Perivascular-derived stem cells with neural crest characteristics are involved in tendon repair. Stem Cells Dev 2015;24(7):857–68.
    doi: 10.1089/scd.2014.0036pmc: PMC4367512pubmed: 25381682google scholar: lookup
  29. Dunkman AA, Buckley MR, Mienaltowski MJ, Adams SM, Thomas SJ, Satchell L, Kumar A, Pathmanathan L, Beason DP, Iozzo RV, Birk DE, Soslowsky LJ. The tendon injury response is influenced by decorin and biglycan. Ann Biomed Eng 2014;42(3):619–30.
    doi: 10.1007/s10439-013-0915-2pmc: PMC3943488pubmed: 24072490google scholar: lookup
  30. Dourte LM, Pathmanathan L, Jawad AF, Iozzo RV, Mienaltowski MJ, Birk DE, Soslowsky LJ. Influence of decorin on the mechanical, compositional, and structural properties of the mouse patellar tendon. J Biomech Eng 2012;134(3):031005.
    doi: 10.1115/1.4006200pmc: PMC3705828pubmed: 22482685google scholar: lookup
  31. Nguyen PK, Jana A, Huang C, Grafton A, Holt I, Giacomelli M, Kuo CK. Tendon mechanical properties are enhanced via recombinant lysyl oxidase treatment. Front Bioeng Biotechnol 2022;10:945639.
    doi: 10.3389/fbioe.2022.945639pmc: PMC9389157pubmed: 35992359google scholar: lookup
  32. Eisner LE, Rosario R, Andarawis-Puri N, Arruda EM. The role of the Non-collagenous Extracellular Matrix in Tendon and Ligament Mechanical Behavior: a review. J Biomech Eng 2022;144(5):050801.
    doi: 10.1115/1.4053086pmc: PMC8719050pubmed: 34802057google scholar: lookup
  33. Steffen D, Mienaltowski MJ, Baar K. Scleraxis and collagen I expression increase following pilot isometric loading experiments in a rodent model of patellar tendinopathy. Matrix Biol 2022;109:34–48.
    doi: 10.1016/j.matbio.2022.03.006pubmed: 35358711google scholar: lookup
  34. Cai L, Xiong X, Kong X, Xie J. The role of the Lysyl oxidases in tissue repair and remodeling: a concise review. Tissue Eng Regen Med 2017;14(1):15–30.
    doi: 10.1007/s13770-016-0007-0pmc: PMC6171574pubmed: 30603458google scholar: lookup
  35. Garcia-Guerra, Vila-Bedmar, Carrasco-Ranso M, Cruces-Sande M, Martin M, Ruiz-Gomez A, Ruiz-Gomez M, Lorenzo M, Fernandez-Veledo S, Mayor F Jr, Murga C, Nieto-Vazquez I. Skeletal muscle myogenesis is regulated by G-coupled receptor kinase 2. J Mol Cell Biol 2014;6(4):299–311.
    doi: 10.1093/jmcb/mju025pubmed: 24927997google scholar: lookup
  36. Liu Z, Hussien AA, Wang Y, Heckmann T, Gonzalez R, Karner CM, Snedeker JG, Gray RS. An adhesion G protein-coupled receptor is required in cartilaginous and dense connective tissues to maintain spine alignment. eLife 2021;10:e67781.
    doi: 10.7554/eLife.67781pmc: PMC8328515pubmed: 34318745google scholar: lookup
  37. Newton AH, Pask AJ. CHD9 upregulates RUNX2 and has a potential role in skeletal evolution. BMC Mol Cell Biol 2020;21(1):27.
    doi: 10.1186/s12860-020-00270-5pmc: PMC7161146pubmed: 32295522google scholar: lookup
  38. Ansorge HL, Meng X, Zhang G, Veit G, Sun M, Klement JF, Beason DP, Soslowsky LJ, Koch M, Birk DE. Type XIV Collagen regulates fibrillogenesis: premature collagen Fibril Growth and tissue dysfunction in null mice. J Biol Chem 2009;284(13):8427–38.
    doi: 10.1074/jbc.M805582200pmc: PMC2659201pubmed: 19136672google scholar: lookup
  39. Ishibashi K, Ikegami K, Shimbo T, Sasaki E, Kitayama T, Nakamura Y, Tsushima T, Ishibashi Y, Tamai K. Single-cell transcriptome analysis reveals cellular heterogeneity in mouse intra- and extra articular ligaments. Commun Biol 2022;12(5):1233.
    doi: 10.1038/s42003-022-04196-wpmc: PMC9653455pubmed: 36371589google scholar: lookup
  40. Srivastava D, Thomas T, Lin Q, Kirby ML, Brown D, Olson EN. Regulation of cardiac mesodermal and neural crest development by the bHLH transcription factor, dHAND. Nat Genet 1997;16(2):154–60.
    doi: 10.1038/ng0697-154pubmed: 9171826google scholar: lookup
  41. Charité J, McFadden DG, Olson EN. The bHLH transcription factor dHAND controls sonic hedgehog expression and establishment of the zone of polarizing activity during limb development. Development 2000;127(11):2461–70.
    doi: 10.1242/dev.127.11.2461pubmed: 10804186google scholar: lookup
  42. Thomas T, Kurihara H, Yamagishi H, Kurihara Y, Yazaki Y, Olson EN, Srivastava D. A signaling cascade involving endothelin-1, dHAND and msx1 regulates development of neural-crest-derived branchial arch mesenchyme. Development 1998;125(16):3005–14.
    doi: 10.1242/dev.125.16.3005pubmed: 9671575google scholar: lookup
  43. Fernandez-Teran M, Piedra ME, Kathiriya IS, Srivastava D, Rodriguez-Rey JC, Ros MA. Role of dHAND in the anterior-posterior polarization of the limb bud: implications for the sonic hedgehog pathway. Development 2000;127(10):2133–42.
    doi: 10.1242/dev.127.10.2133pubmed: 10769237google scholar: lookup
  44. McFadden DG, McAnally J, Richardson JA, Charité J, Olson EN. Misexpression of dHAND induces ectopic digits in the developing limb bud in the absence of direct DNA binding. Development 2002;129(13):3077–88.
    doi: 10.1242/dev.129.13.3077pubmed: 12070084google scholar: lookup
  45. DeLaurier A, Schweitzer R, Logan M. Pitx1 determines the morphology of muscle, tendon, and bones of the hindlimb. Dev Biol 2006;299(1):22–34.
    doi: 10.1016/j.ydbio.2006.06.055pubmed: 16989801google scholar: lookup
  46. Stolle K, Schnoor, Fuellen G, Spitzer M, Cullen P, Lorkowski S. Cloning, genomic organization, and tissue-specific expression of the RASL11B gene. Biochim Biophys Acta 2007;1769(7–8):514–24.
    doi: 10.1016/j.bbaexp.2007.05.005pubmed: 17628721google scholar: lookup
  47. Luo Y, Wang A-T, Zhang Q-F, Liu R-M, Xiao J-H. RASL11B gene enhances hyaluronic acid-mediated chondrogenic differentiation in human amniotic mesenchymal stem cells via the activation of Sox9/ERK/smad signals. Exp Biol Med 2020;245(18):1708–21.
    doi: 10.1177/1535370220944375pmc: PMC7802383pubmed: 32878463google scholar: lookup
  48. Berthet E, Chen C, Butcher K, Schneider RA, Alliston T, Amirtharajah M. Smad3 binds Scleraxis and Mohawk and regulated tendon development. J Orthop Res 2013;31(9):1475–83.
    doi: 10.1002/jor.22382pmc: PMC3960924pubmed: 23653374google scholar: lookup
  49. Arnesen SM, Lawson MA. Age-related changes in focal adhesions lead to altered cell behavior in tendon fibroblasts. Mech Ageing Dev 2006;127:726–32.
    doi: 10.1016/j.mad.2006.05.003pubmed: 16780927google scholar: lookup
  50. Bailey AJ, Paul RG, Knott L. Mechanisms of maturation and ageing of collagen. Mech Ageing Dev 1998;106:1–56.
    doi: 10.1016/S0047-6374(98)00119-5pubmed: 9883973google scholar: lookup
  51. Tsai WC, Chang HN, Yu TY, Chien CH, Fu LF, Liang FC, Pang JH. Decreased proliferation of aging tenocytes is associated with down-regulation of cellular senescence-inhibited gene and up-regulation of p27. J Orthop Res 2011;29:1598–603.
    doi: 10.1002/jor.21418pubmed: 21452304google scholar: lookup
  52. Connizzo BK, Sarver JJ, Birk DE, Soslowsky LJ, Iozzo RV. Effect of age and proteoglycan deficiency on collagen fiber re-alignment and mechanical properties in mouse supraspinatus tendon. J Biomech Eng 2013;135:021019.
    doi: 10.1115/1.4023234pmc: PMC5413158pubmed: 23445064google scholar: lookup
  53. Wu PT, Su WR, Li CL, Hsieh JL, Ma CH, Wu CL, Kuo LC, Jou IM, Chen SY. Inhibition of CD44 induces apoptosis, inflammation, and matrix metalloproteinase expression in tendinopathy. J Biol Chem 2019;294(52):20177–84.
    doi: 10.1074/jbc.RA119.009675pmc: PMC6937571pubmed: 31732563google scholar: lookup
  54. Smith RK, Gerard M, Dowling B, Dart AJ, Birch HL, Goodship AE. Correlation of cartilage oligomeric matrix protein (COMP) levels in equine tendon with mechanical properties: a proposed role for COMP in determining function-specific mechanical characteristics of locomotor tendons. Equine Vet J Suppl 2002; (34):241–4.
    pubmed: 12405694
  55. Dai G, Li Y, Liu J, Zhang C, Chen M, Lu P, Rui Y. Higher BMP expression in Tendon Stem/Progenitor cells contributes to the increased heterotopic ossification in Achilles Tendon with Aging. Front Cell Dev Biol 2020;8:570605.
    doi: 10.3389/fcell.2020.570605pmc: PMC7546413pubmed: 33102476google scholar: lookup
  56. Unnikrishnan A, Hadad N, Masser DR, Jackson J, Freeman WM, Richardson A. Revisiting the genomic hypomethylation hypothesis of aging. Ann N Y Acad Sci 2018;1418(1):69–79.
    doi: 10.1111/nyas.13533pmc: PMC5934293pubmed: 29363785google scholar: lookup
  57. Besselink N, Keijer J, Vermeulen C, Boymans S, de Ridder J, van Hoeck A, Cuppen E, Kuijk E. The genome-wide mutational consequences of DNA hypomethylation. Sci Rep 2023;13(1):6874.
    doi: 10.1038/s41598-023-33932-3pmc: PMC10140063pubmed: 37106015google scholar: lookup
  58. Riasat K, Bardell D, Goljanek-Whysall K, Clegg PD, Peffers MJ. Epigenetic mechanisms in Tendon Ageing. Br Med Bull 2020;135(1):90–107.
    doi: 10.1093/bmb/ldaa023pmc: PMC7585832pubmed: 32827252google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,103 horse owners like you who receive our email updates on horse health and nutrition.