Abstract
Osteochondrosis (OC) is a developmental disease in horses with a significant impact on the horse’s welfare and performance. Previously, differentially expressed genes in leukocytes of OC-affected have been identified and were differentially expressed in horses of different ages when compared to their age-matched controls.As the time course of the development of OC lesions seems to be joint dependent,the aim of this study is to compare in young OCaffected horses (between 8 to 12 months), the different expression of selected genes depending the joints involved.The expression of OC-related genes were analysed by rt-PCR and subsequent statistical analysis (ΔΔCT) in the leukocytes of 30 Belgian Warmblood horses aged between 8 to 12 months divided in groups depending the affected joints (fetlock, hock and stifle).In the three groups, expression of ApoB-3G, MGAT4A, B4GALT6 and PRKCG genes were significantly higher in the OC-affected foals compared to the healthy foals. Based on the profiles of expression ofApoB-3G, Dsh1/Dvl1, Foxl1, Hp, ISG15, Mark2, PPR2A, RUSC2 and WASH1 genes,the localization of the disease can be determined: expression levels of ApoB3G, WASH1 and FOXl1 to identify fetlock, ApoB3G, PPR2A to identify OC-development in the hock and ApoB3G, Dsh1/Dvl1, WASH1, PPP2R1A and Mark2 geneto identify OC-development in the stifle. However at this moment, the rt-PCR analysis of the identified genes as biomarkers gives only diagnostic information. For the future, the profile of expression of these genes could give also some predictive information on the evolution of the disease such as remission or permanent OC-lesions.
Highlights
The developmental orthopaedic disease osteochondrosis (OC) affects growing horses and severely compromise their athletic careers [1,2,3,4]
We have identified 2,553 genes significantlyup or down regulated between the OC-group and the control group [17].For the present study, we chosen35genes following different criteria: a low p-value, fold induction superior to 5, quality of annotation, a mix of up- and down-regulated genes involved in known signalling pathways (Table 1).Four control genes: WARS (Tryptophanyl-tRNAsynthetase), RIGE (Retinoic acid-induced gene E protein), B2M(Beta-2-microglobulin) and TUBB2C (Tubulin, beta 2C), referred to as housekeeping genes, were used to normalise mRNA levels between different samples.Real-time PCR analysis was performed for each of the markers and housekeeping genes, for each measured marker or housekeeping gene, a cycle threshold value (Ctvalue) was obtained
We identified localization-dependent differentially expressed genes in leukocytes of OC-affected horses
Summary
The developmental orthopaedic disease osteochondrosis (OC) affects growing horses and severely compromise their athletic careers [1,2,3,4]. The disease appears to be multifactorial in origin, including skeletal growth rates, nutrition, endocrinologic factors, and exercise of the horse, biomechanics, and genetic effects [10]. High energy diet is known to induce OC lesions in foals [11,12]. It appears that mares fed with concentrates during gestation are more likely to produce foals that are subsequently affected by OC compared with other mares [13]
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