Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
Kinetics of Gene Expression Changes in Equine Fetal Interzone and Anlagen Cells Over 14 Days of Induced Chondrogenesis.
Abstract: Within developing synovial joints, interzone and anlagen cells progress through divergent chondrogenic pathways to generate stable articular cartilage and transient hypertrophic anlagen cartilage, respectively. Understanding the comparative cell biology between interzone and anlagen cells may provide novel insights into emergent cell-based therapies to support articular cartilage regeneration. The aim of this study was to assess the kinetics of gene expression profiles in these skeletal cell lines after inducing chondrogenesis in culture. Interzone and anlagen cells from seven equine fetuses were isolated and grown in a TGF-β1 chondrogenic inductive medium. Total RNA was isolated at ten time points (0, 1.5, 3, 6, 12, 24, 48, 96, 168, and 336 h), and gene expression for 93 targeted gene loci was measured in a microfluidic RT-qPCR system. Differential transcriptional responses were observed as early as 1.5 h after the initiation of chondrogenesis. Genes with functional annotations that include transcription regulation responded to the chondrogenic stimulation earlier (1.5-96 h) than genes involved in signal transduction (1.5-336 h) and the extracellular matrix biology (3-336 h). Between interzone and anlagen cell cultures, expression levels of 73 out of the 93 targeted genes were not initially different at 0 h, but 47 out of the 73 genes became differentially expressed under the chondrogenic stimulation. While interzone and anlagen cells are both chondrogenic, they display clear differences in response to the same TGF-β1 chondrogenic stimulation. This study provides new molecular insight into a timed sequence of the divergent developmental fates of interzone and anlagen cells in culture over 14 days.
Copyright © 2021 Mok and MacLeod.
Publication Date: 2021-08-09 PubMed ID: 34434986PubMed Central: PMC8380811DOI: 10.3389/fvets.2021.722324Google 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.
- 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.
This research aimed to understand the difference in gene expression between equine fetal interzone and anlagen cells when undergoing transition into cartilage. Studying such changes might shed light on novel cell-based therapies for cartilage regeneration.
Research Design
- The goal of this study was to analyze the changes or kinetics in gene expression profiles when interzone and anlagen cells undergo chondrogenesis.
- Cells from these two sources were collected from seven equine fetuses and cultured in a TGF-β1 conductive medium designed to stimulate chondrogenesis.
- Researchers harvested total RNA at ten different time intervals following the initiation of chondrogenesis to analyze gene expression.
Findings
- Shifts in gene expression began to appear as soon as 1.5 hours after the start of chondrogenesis.
- Genes associated with transcription regulation responded to the chondrogenic stimulation earlier, followed by genes involved in signal transduction and then those related to the extracellular matrix biology.
- Although interzone and anlagen cells both have the potential to become cartilage, their responses to the same TGF-β1 chondrogenic trigger were distinctly different.
- Out of the 93 targeted genes, initial expression levels at the start (0 hour) were identical for 73 genes in both cell types. But under chondrogenic stimulation, expression levels for 47 of these genes changed differently.
Conclusion
- The study outlined a timed sequence of the developmental differences between interzone and anlagen cells when grown in culture over a 14-day period.
- Understanding the unique gene expressions and responses of these two cell types to chondrogenic stimulation provides a starting point for future research into cell-based treatments for cartilage regeneration.
Cite This Article
APA
Mok CH, MacLeod JN.
(2021).
Kinetics of Gene Expression Changes in Equine Fetal Interzone and Anlagen Cells Over 14 Days of Induced Chondrogenesis.
Front Vet Sci, 8, 722324.
https://doi.org/10.3389/fvets.2021.722324 Publication
Researcher Affiliations
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY, United States.
- Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY, United States.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
This article includes 38 references
- Pitsillides AA, Ashhurst DE. A critical evaluation of specific aspects of joint development.. Dev Dyn 2008 Sep;237(9):2284-94.
- Decker RS, Koyama E, Pacifici M. Genesis and morphogenesis of limb synovial joints and articular cartilage.. Matrix Biol 2014 Oct;39:5-10.
- Alford JW, Cole BJ. Cartilage restoration, part 1: basic science, historical perspective, patient evaluation, and treatment options.. Am J Sports Med 2005 Feb;33(2):295-306.
- Adam EN, Janes J, Lowney R, Lambert J, Thampi P, Stromberg A, MacLeod JN. Chondrogenic differentiation potential of adult and fetal equine cell types.. Vet Surg 2019 Apr;48(3):375-387.
- McCarthy HE, Bara JJ, Brakspear K, Singhrao SK, Archer CW. The comparison of equine articular cartilage progenitor cells and bone marrow-derived stromal cells as potential cell sources for cartilage repair in the horse.. Vet J 2012 Jun;192(3):345-51.
- Rakic R, Bourdon B, Demoor M, Maddens S, Saulnier N, Galéra P. Differences in the intrinsic chondrogenic potential of equine umbilical cord matrix and cord blood mesenchymal stromal/stem cells for cartilage regeneration.. Sci Rep 2018 Sep 14;8(1):13799.
- Franco DL, Mainez J, Vega S, Sancho P, Murillo MM, de Frutos CA, Del Castillo G, López-Blau C, Fabregat I, Nieto MA. Snail1 suppresses TGF-beta-induced apoptosis and is sufficient to trigger EMT in hepatocytes.. J Cell Sci 2010 Oct 15;123(Pt 20):3467-77.
- Aomatsu K, Arao T, Sugioka K, Matsumoto K, Tamura D, Kudo K, Kaneda H, Tanaka K, Fujita Y, Shimomura Y, Nishio K. TGF-β induces sustained upregulation of SNAI1 and SNAI2 through Smad and non-Smad pathways in a human corneal epithelial cell line.. Invest Ophthalmol Vis Sci 2011 Apr 14;52(5):2437-43.
- Ranganathan P, Agrawal A, Bhushan R, Chavalmane AK, Kalathur RK, Takahashi T, Kondaiah P. Expression profiling of genes regulated by TGF-beta: differential regulation in normal and tumour cells.. BMC Genomics 2007 Apr 11;8:98.
- Rudini N, Felici A, Giampietro C, Lampugnani M, Corada M, Swirsding K, Garrè M, Liebner S, Letarte M, ten Dijke P, Dejana E. VE-cadherin is a critical endothelial regulator of TGF-beta signalling.. EMBO J 2008 Apr 9;27(7):993-1004.
- Koyama N, Miura M, Nakao K, Kondo E, Fujii T, Taura D, Kanamoto N, Sone M, Yasoda A, Arai H, Bessho K, Nakao K. Human induced pluripotent stem cells differentiated into chondrogenic lineage via generation of mesenchymal progenitor cells.. Stem Cells Dev 2013 Jan 1;22(1):102-13.
- Yokota J, Chosa N, Sawada S, Okubo N, Takahashi N, Hasegawa T, Kondo H, Ishisaki A. PDGF-induced PI3K-mediated signaling enhances the TGF-β-induced osteogenic differentiation of human mesenchymal stem cells in a TGF-β-activated MEK-dependent manner.. Int J Mol Med 2014 Mar;33(3):534-42.
- Nejadnik H, Diecke S, Lenkov OD, Chapelin F, Donig J, Tong X, Derugin N, Chan RC, Gaur A, Yang F, Wu JC, Daldrup-Link HE. Improved approach for chondrogenic differentiation of human induced pluripotent stem cells.. Stem Cell Rev Rep 2015 Apr;11(2):242-53.
- Yamazaki S, Nakano N, Honjo A, Hara M, Maeda K, Nishiyama C, Kitaura J, Ohtsuka Y, Okumura K, Ogawa H, Shimizu T. The Transcription Factor Ehf Is Involved in TGF-β-Induced Suppression of FcεRI and c-Kit Expression and FcεRI-Mediated Activation in Mast Cells.. J Immunol 2015 Oct 1;195(7):3427-35.
- Adam EN, Zhang Y, Liu X, Coleman S, Flight R, Lambert J. Gene expression patterns that distinguish equine articular cartilage from other cartilaginous tissues.. Presentation Abstracts, 2017 ACVS Surgery Summit (2017) 46:E1–75.
- Mienaltowski MJ, Huang L, Stromberg AJ, MacLeod JN. Differential gene expression associated with postnatal equine articular cartilage maturation.. BMC Musculoskelet Disord 2008 Nov 5;9:149.
- Hestand MS, Kalbfleisch TS, Coleman SJ, Zeng Z, Liu J, Orlando L, MacLeod JN. Annotation of the Protein Coding Regions of the Equine Genome.. PLoS One 2015;10(6):e0124375.
- Fluidigm . Real-Time PCR Analysis PN, 68000088 N1 User Guide. South San Francisco CA: Fluidigm; (2018).
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001 Dec;25(4):402-8.
- Yamamoto KR, Alberts BM. Steroid receptors: elements for modulation of eukaryotic transcription.. Annu Rev Biochem 1976;45:721-46.
- Eser P, Demel C, Maier KC, Schwalb B, Pirkl N, Martin DE, Cramer P, Tresch A. Periodic mRNA synthesis and degradation co-operate during cell cycle gene expression.. Mol Syst Biol 2014;10(1):717.
- Bi W, Deng JM, Zhang Z, Behringer RR, de Crombrugghe B. Sox9 is required for cartilage formation.. Nat Genet 1999 May;22(1):85-9.
- Akiyama H, Chaboissier MC, Martin JF, Schedl A, de Crombrugghe B. 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 2002 Nov 1;16(21):2813-28.
- Ninomiya K, Miyamoto T, Imai J, Fujita N, Suzuki T, Iwasaki R, Yagi M, Watanabe S, Toyama Y, Suda T. Osteoclastic activity induces osteomodulin expression in osteoblasts.. Biochem Biophys Res Commun 2007 Oct 19;362(2):460-6.
- Mevel R, Draper JE, Lie-A-Ling M, Kouskoff V, Lacaud G. RUNX transcription factors: orchestrators of development.. Development 2019 Sep 5;146(17).
- Kwan AP, Cummings CE, Chapman JA, Grant ME. Macromolecular organization of chicken type X collagen in vitro.. J Cell Biol 1991 Aug;114(3):597-604.
- Schmid TM, Cole AA, Chen Q, Bonen DK, Luchene L, Linsenmayer TF. Assembly of Type X collagen by hypertrophic chondrocytes.. Extracellular Matrix Assembly and Structure San Diego: Academic Press; (1994). p. 171–206.
- Luo G, Ducy P, McKee MD, Pinero GJ, Loyer E, Behringer RR, Karsenty G. Spontaneous calcification of arteries and cartilage in mice lacking matrix GLA protein.. Nature 1997 Mar 6;386(6620):78-81.
- Settle SH Jr, Rountree RB, Sinha A, Thacker A, Higgins K, Kingsley DM. Multiple joint and skeletal patterning defects caused by single and double mutations in the mouse Gdf6 and Gdf5 genes.. Dev Biol 2003 Feb 1;254(1):116-30.
- Krejci P, Krakow D, Mekikian PB, Wilcox WR. Fibroblast growth factors 1, 2, 17, and 19 are the predominant FGF ligands expressed in human fetal growth plate cartilage.. Pediatr Res 2007 Mar;61(3):267-72.
- Hyde G, Dover S, Aszodi A, Wallis GA, Boot-Handford RP. Lineage tracing using matrilin-1 gene expression reveals that articular chondrocytes exist as the joint interzone forms.. Dev Biol 2007 Apr 15;304(2):825-33.
- Golub EE, Boesze-Battaglia K. The role of alkaline phosphatase in mineralization.. Curr Opin Orthop (2007) 18, 444–8.
- Hodgkinson CP, Naidoo V, Patti KG, Gomez JA, Schmeckpeper J, Zhang Z, Davis B, Pratt RE, Mirotsou M, Dzau VJ. Abi3bp is a multifunctional autocrine/paracrine factor that regulates mesenchymal stem cell biology.. Stem Cells 2013 Aug;31(8):1669-82.
- Hissnauer TN, Baranowsky A, Pestka JM, Streichert T, Wiegandt K, Goepfert C, Beil FT, Albers J, Schulze J, Ueblacker P, Petersen JP, Schinke T, Meenen NM, Pörtner R, Amling M. Identification of molecular markers for articular cartilage.. Osteoarthritis Cartilage 2010 Dec;18(12):1630-8.
- Pfeifer A, Aszódi A, Seidler U, Ruth P, Hofmann F, Fässler R. Intestinal secretory defects and dwarfism in mice lacking cGMP-dependent protein kinase II.. Science 1996 Dec 20;274(5295):2082-6.
- Kawasaki Y, Kugimiya F, Chikuda H, Kamekura S, Ikeda T, Kawamura N, Saito T, Shinoda Y, Higashikawa A, Yano F, Ogasawara T, Ogata N, Hoshi K, Hofmann F, Woodgett JR, Nakamura K, Chung UI, Kawaguchi H. Phosphorylation of GSK-3beta by cGMP-dependent protein kinase II promotes hypertrophic differentiation of murine chondrocytes.. J Clin Invest 2008 Jul;118(7):2506-15.
- Bond SR, Lau A, Penuela S, Sampaio AV, Underhill TM, Laird DW, Naus CC. Pannexin 3 is a novel target for Runx2, expressed by osteoblasts and mature growth plate chondrocytes.. J Bone Miner Res 2011 Dec;26(12):2911-22.
- Oh SK, Shin JO, Baek JI, Lee J, Bae JW, Ankamerddy H, Kim MJ, Huh TL, Ryoo ZY, Kim UK, Bok J, Lee KY. Pannexin 3 is required for normal progression of skeletal development in vertebrates.. FASEB J 2015 Nov;29(11):4473-84.
Citations
This article has been cited 1 times.- Menarim BC, Mok CH, Scoggin KE, Gornik A, Adam EN, Loux SC, MacLeod JN. Fetal Cartilage Progenitor Cells in the Repair of Osteochondral Defects. JB JS Open Access 2025 Jan-Mar;10(1).
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
