Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney

<div><p>The aim of this study was to determine the relative abundance and relationship of vitamin D responsive and calcium transporting transcripts (TRPV5, TRPV6, calD_9k, calD_28k, PMCA, NCX1, CYP27B1, CYP24A1, and VDR) in ovine, canine and, equine kidney using quantitative real-time PCR (RT-qPCR), and then perform a comparison between the three species. Renal tissue samples were harvested post-mortem from 10 horses, 10 sheep, and five dogs. Primers were designed for each gene. For each sample total RNA was extracted, cDNA synthesised, and RT-qPCR was performed. RT-qPCR data were normalised and statistical comparison was performed. Due to their consistent correlation with each other in each species, TRPV6, calD_9k/calD_28k, and PMCA appeared to be the main pathways involved in active transepithelial calcium transport in the kidney of sheep, dogs and horses. The results indicate that all of the studied genes were expressed in the renal tissue of studied species, although the expression levels and correlation of transcripts with each other were different from species to species. All vitamin D responsive and calcium transporting transcripts were highly correlated with VDR in equine kidney, but not in sheep and dogs. The CYP27B1 and CYP24A1 mRNAs showed a different renal expression pattern and correlation in horses compared with sheep and dogs. Given the high urinary calcium concentration and low serum 1,25(OH)₂D concentration in horses, it could be expected that CYP27B1 expression would be lower than CYP24A1 in the horse, and this did not appear to be the case. The findings suggest that despite low serum vitamin D concentrations, vitamin D still plays a significant role in calcium metabolism in horses, especially given the strong correlations between VDR and vitamin D responsive transcripts in these animals.</p></div

LMNSNPdata

This data achieve including all the genotypes for each SNP for each animal. The files were created from the original genotyping file using "awk" or "sort" commands

Correlation between the relative expression of Forkhead box transcription factor (FoxO1) and osteotesticular protein tyrosine phosphatase (Esp) mRNA.

<p>The expression of FoxO1 and Esp were significantly correlated (R=0.59, p=0.0006). rlog FoxO1 = residuals of log of FoxO1 relative expression. rlog Esp = residuals of log of Esp relative expression.</p

Antilog of relative expression of osteocalcin (OC), insulin receptor (InsR), Forkhead box transcription factor O1 (FoxO1), and osteotesticular protein tyrosine phosphatase (Esp) mRNA.

<p>There were no significant differences in the relative expression of these genes in the offspring of exercised and control dams. Males expressed approximately twice as much Esp mRNA as females (p=0.007). Error bars are antilog of mean ± SE on a log scale (as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082378#pone-0082378-t005" target="_blank">Table 5</a>).</p

Correlation between the relative expression of Forkhead box transcription factor (FoxO1) and osteocalcin (OC) mRNA.

<p>The expression of FoxO1 and OC were significantly correlated (R=0.79, p&lt;0.0001). rlog FoxO1 = residuals of log of FoxO1 relative expression. rlog OC = residuals of log of OC relative expression.</p