Identification of Novel Equine (Equus caballus) Tendon Markers Using RNA Sequencing.
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 (EYA2) and a G-protein regulated inducer of neurite outgrowth 3 (GPRIN3) as specific tendon markers using RT-qPCR. Equine tendon cells cultured as three-dimensional spheroids expressed significantly greater levels ofEYA2thanGPRIN3, 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 identifiedEYA2andGPRIN3as 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.
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.
Jan M
JM
Kuemmerle
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.
Felix
F
Theiss
Scientific IT Services, Swiss Federal Institute of Technology, CH 8092 Zurich, Switzerland. michalo@id.ethz.ch.
Michal J
MJ
Okoniewski
Institute of Laboratory Animal Science, University of Zurich, CH-8057 Zurich, Switzerland. fabienne.weber@lasc.uzh.ch.
Fabienne A
FA
Weber
Division of Trauma Surgery, Center for Clinical Research, University Hospital Zurich, University of Zurich, Sternwartstrasse 14, CH-8091 Zurich, Switzerland. Sonja.hemmi@usz.ch.
Sonja
S
Hemmi
Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. ali.mirsaidi@cabmm.uzh.ch.
Ali
A
Mirsaidi
Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland. peter.richards@cabmm.uzh.ch.
Peter J
PJ
Richards
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.
Paolo
P
Cinelli
G-protein regulated inducer of neurite outgrowth 3; eyes absent homolog 2; horse; tendon; transcriptome