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Genes2016; 7(11); 97; doi: 10.3390/genes7110097

Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing.

Abstract: Although several tendon-selective genes exist, they are also expressed in other musculoskeletal tissues. As cell and tissue engineering is reliant on specific molecular markers to discriminate between cell types, tendon-specific genes need to be identified. In order to accomplish this, we have used RNA sequencing (RNA-seq) to compare gene expression between tendon, bone, cartilage and ligament from horses. We identified several tendon-selective gene markers, and established eyes absent homolog 2 () and a G-protein regulated inducer of neurite outgrowth 3 () as specific tendon markers using RT-qPCR. Equine tendon cells cultured as three-dimensional spheroids expressed significantly greater levels of than , and stained positively for EYA2 using immunohistochemistry. EYA2 was also found in fibroblast-like cells within the tendon tissue matrix and in cells localized to the vascular endothelium. In summary, we have identified and as specific molecular markers of equine tendon as compared to bone, cartilage and ligament, and provide evidence for the use of EYA2 as an additional marker for tendon cells in vitro.
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Publication Date: 2016-11-10 PubMed ID: 27834918PubMed Central: PMC5126783DOI: 10.3390/genes7110097Google Scholar: Lookup
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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 article focuses on identifying specific molecular markers for equine (horse) tendons using RNA sequencing, to better distinguish them from other musculoskeletal tissues. This is key to cell and tissue engineering that seek to understand specific cell types. The researchers found that the genes eyes absent homolog 2 (EYA2) and a G-protein regulated inducer of neurite outgrowth 3 (GPRIN3) can be used as specific tendon markers.

RNA Sequencing and the Search for Tendon Markers

  • The research began with the challenge that, although several genes are specifically expressed in tendons, these same genes also show expression in other musculoskeletal tissues. This overlap makes it difficult for researchers to accurately discriminate between cell types in cellular and tissue engineering studies.
  • To solve this issue, the researchers employed RNA sequencing (RNA-seq), a powerful tool for studying gene expression. They compared the gene expression profiles of tendon, bone, cartilage, and ligament tissues harvested from horses.

Identification of Tendon-Specific Markers

  • Through this process of gene comparison, researchers successfully identified several gene markers that are selectively expressed in tendons. In particular, two genes, EYA2 and GPRIN3, stood out as unique and specific tendon markers.
  • After their identification via RNA-seq, these markers were further confirmed using RT-qPCR, a technique used to measure the quantity of specific RNAs (in this case, EYA2 and GPRIN3).

Verification of Tendon Markers and Their Implications

  • The team also found that equine tendon cells, when cultured in a three-dimensional environment, expressed significantly more EYA2 than GPRIN3. This observation was corroborated by immunohistochemistry staining, which also confirmed the presence of EYA2 in the cultured cells.
  • EYA2 was also detected in fibroblast-like cells (a cell type commonly found in tendons) within the tendon tissue matrix, as well as in cells situated in the vascular endothelium. This indicates EYA2’s potential for being a useful marker to identify and study tendon cells both in vivo and in vitro.
  • In conclusion, the identification of EYA2 and GPRIN3 as specific molecular markers for equine tendons represents a significant advance in the field of tissue engineering and can facilitate further studies into understanding the biology of tendons.

Cite This Article

APA
Kuemmerle JM, Theiss F, Okoniewski MJ, Weber FA, Hemmi S, Mirsaidi A, Richards PJ, Cinelli P. (2016). Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing. Genes (Basel), 7(11), 97. https://doi.org/10.3390/genes7110097

Publication

Genes
ISSN: 2073-4425
NlmUniqueID: 101551097
Country: Switzerland
Language: English
Volume: 7
Issue: 11
PII: 97

Researcher Affiliations

Kuemmerle, Jan M
  • Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. jkuemmerle@vetclinics.uzh.ch.
  • Equine Hospital, Vetsuisse Faculty, University of Zurich, CH-8057 Zurich, Switzerland. jkuemmerle@vetclinics.uzh.ch.
Theiss, Felix
  • Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. ftheiss@vetclinics.uzh.ch.
  • Equine Hospital, Vetsuisse Faculty, University of Zurich, CH-8057 Zurich, Switzerland. ftheiss@vetclinics.uzh.ch.
Okoniewski, Michal J
  • Scientific IT Services, Swiss Federal Institute of Technology, CH 8092 Zurich, Switzerland. michalo@id.ethz.ch.
Weber, Fabienne A
  • Institute of Laboratory Animal Science, University of Zurich, CH-8057 Zurich, Switzerland. fabienne.weber@lasc.uzh.ch.
Hemmi, Sonja
  • Division of Trauma Surgery, Center for Clinical Research, University Hospital Zurich, University of Zurich, Sternwartstrasse 14, CH-8091 Zurich, Switzerland. Sonja.hemmi@usz.ch.
Mirsaidi, Ali
  • Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. ali.mirsaidi@cabmm.uzh.ch.
Richards, Peter J
  • Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. peter.richards@cabmm.uzh.ch.
Cinelli, Paolo
  • Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. paolo.cinelli@usz.ch.
  • Division of Trauma Surgery, Center for Clinical Research, University Hospital Zurich, University of Zurich, Sternwartstrasse 14, CH-8091 Zurich, Switzerland. paolo.cinelli@usz.ch.

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

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