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Home / Equine Research Bank / PLoS One / Evaluation and Comparison of Vitamin D Responsive Gene Expre...
PloS one2016; 11(9); e0162598; doi: 10.1371/journal.pone.0162598

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

Abstract: The aim of this study was to determine the relative abundance and relationship of vitamin D responsive and calcium transporting transcripts (TRPV5, TRPV6, calD9k, calD28k, 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, calD9k/calD28k, 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.
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Publication Date: 2016-09-15 PubMed ID: 27632366PubMed Central: PMC5025205DOI: 10.1371/journal.pone.0162598Google 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 article presents research looking at how genes respond to Vitamin D in ovine (sheep), canine (dogs), and equine (horses) kidneys. It found differences among the species, but indicated that for all, Vitamin D plays a role in regulating calcium.

Research Background and Methodology

  • Researchers aimed to examine and compare gene expressions responsive to vitamin D in kidneys across three species: sheep, dogs, and horses.
  • Vitamin D is known for regulating calcium, which is crucial in bone growth and functioning of the nervous system. The genes of interest were those linked with vitamin D’s activity, and those related to calcium transporting.
  • 10 horses, 10 sheep, and five dogs were part of this study. The researchers collected kidney tissue samples from these animals post-mortem.
  • The researchers extracted total RNA from these samples, synthesized cDNA, and then conducted quantitative real-time PCR (RT-qPCR), which amplifies and simultaneously quantifies targeted DNA molecules.
  • RT-qPCR data were normalized (adjusted to even out any bias in the data) for statistical comparisons.

Main Findings

  • According to the results, all of the studied vitamin D responsive and calcium transporting genes were highly correlated with Vitamin D Receptor (VDR) in horse kidneys—although this correlation was not found in sheep and dogs.
  • TRPV6, calD9k/calD28k, and PMCA transcripts, which were shown to exhibit consistent correlation with each other in every evaluated species, were found to be the primary pathways involved in transepithelial calcium transport in the kidneys horses, sheep, and dogs.
  • There was variation in the expression levels of these genes between species, emphasizing the potential differences in vitamin D and calcium regulation as per species.
  • The researchers also noticed that even though horses generally have high urinary calcium concentration and low serum 1,25(OH)2D concentration, the expression of key gene, CYP27B1 was not less than the expression of CYP24A1 in horse kidneys, which was not as expected.

Conclusion and Implication of Findings

  • The study concludes by emphasizing the critical role of vitamin D in calcium metabolism in horses. This finding is significant considering horses’ typically low serum vitamin D concentrations but strong correlations between VDR and vitamin D responsive transcripts.
  • The inter-species variation in gene expression further provides important insight into how vitamin D may exert different influences on calcium metabolism in different species.

Cite This Article

APA
Azarpeykan S, Dittmer KE, Marshall JC, Perera KC, Gee EK, Acke E, Thompson KG. (2016). Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney. PLoS One, 11(9), e0162598. https://doi.org/10.1371/journal.pone.0162598

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 11
Issue: 9
Pages: e0162598

Researcher Affiliations

Azarpeykan, Sara
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.
Dittmer, Keren E
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.
Marshall, Jonathan C
  • Institute of Fundamental Sciences (IFS), Massey University, Palmerston North, New Zealand.
Perera, Kalyani C
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.
Gee, Erica K
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.
Acke, Els
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.
Thompson, Keith G
  • Institute of Veterinary, Animal and Biomedical Science (IVABS), Massey University, Palmerston North, New Zealand.

MeSH Terms

  • Animals
  • Dogs
  • Gene Expression Regulation / drug effects
  • Horses
  • Kidney / metabolism
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Sheep
  • TRPV Cation Channels / genetics
  • TRPV Cation Channels / metabolism
  • Vitamin D / pharmacology

Conflict of Interest Statement

The authors have declared that no competing interests exist.

References

This article includes 48 references
  1. Hoenderop JGJ, van der Kemp A, Hartog A, van de Graaf SFJ, van Os CH, Willems P. Molecular identification of the apical Ca2+ channel in 1,25-dihydroxyvitamin D3-responsive epithelia.. J Biol Chem 1999; 274(13):8375–8.
    pubmed: 10085067
  2. Alexander RT, Rievaj J, Dimke H. Paracellular calcium transport across renal and intestinal epithelia.. Biochemistry and Cell Biology-Biochimie Et Biologie Cellulaire 2014; 92(6):467–80.
    doi: 10.1139/bcb-2014-0061pubmed: 25386841google scholar: lookup
  3. Hoenderop JGJ, Nilius B, Bindels RJM. Calcium absorption across epithelia.. Physiol Rev 2005; 85(1):373–422.
    pubmed: 15618484
  4. Blaine J, Chonchol M, Levi M. Renal Control of Calcium, Phosphate, and Magnesium Homeostasis.. Clin J Am Soc Nephrol 2015; 10(7):1257–72.
    doi: 10.2215/CJN.09750913pmc: PMC4491294pubmed: 25287933google scholar: lookup
  5. Pak CYC, Kaplan R, Bone H, Townsend J, Waters O. A simple test for the diagnosis of absorptive, resorptive and renal hypercalciurias.. N Engl J Med 1975; 292(10):497–500.
    pubmed: 163960
  6. Dittmer KE, Thompson KG. Vitamin D Metabolism and Rickets in Domestic Animals: A Review.. Vet Pathol 2011; 48(2):389–407.
    doi: 10.1177/0300985810375240pubmed: 20634407google scholar: lookup
  7. Radlovic N, Mladenovic M, Simic D, Radlovic P. Vitamin D in the Light of Current Knowledge.. Srp Arh Celok Lek 2012; 140(1–2):110–4.
    pubmed: 22462359
  8. Dusso AS, Brown AJ, Slatopolsky E. Vitamin D.. American Journal of Physiology-Renal Physiology 2005; 289(1):8–28.
    pubmed: 15951480
  9. Kumar R, Tebben PJ, Thompson JR. Vitamin D and the kidney.. Arch Biochem Biophys 2012; 523(1):77–86.
    doi: 10.1016/j.abb.2012.03.003pmc: PMC3361542pubmed: 22426203google scholar: lookup
  10. Adams JS, Hewison M. Update in Vitamin D.. J Clin Endocrinol Metab 2010; 95(2):471–8.
    doi: 10.1210/jc.2009-1773pmc: PMC2840860pubmed: 20133466google scholar: lookup
  11. van de Graaf SFJ, Bindels RJM, Hoenderop JGJ. Physiology of epithelial Ca2+ and Mg2+ transport.. Reviews of Physiology Biochemistry and Pharmacology 2007. p. 77–160.
    pubmed: 17729442
  12. Kayis SA, Atli MO, Kurar E, Bozkaya F, Semacan A, Aslan S. Rating of putative housekeeping genes for quantitative gene expression analysis in cyclic and early pregnant equine endometrium.. Anim Reprod Sci 2011; 125(1–4):124–32.
    doi: 10.1016/j.anireprosci.2011.02.019pubmed: 21411251google scholar: lookup
  13. Peters IR, Peeters D, Helps CR, Day MJ. Development and application of multiple internal reference (housekeeper) gene assays for accurate normalisation of canine gene expression studies.. Vet Immunol Immunopathol 2007; 117(1–2):55–66.
    pubmed: 17346803
  14. Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J. qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data.. Genome Biol 2007; 8(2).
    pmc: PMC1852402pubmed: 17291332
  15. R Development CoreTeam. R: A language and environment for statistical computing. Vienna, Austria: R Foundation for Statistical Computing; 2014.
  16. Bates D, Mächler M, Bolker B, Walker S. Fitting linear mixed-effects models using lme4.. Journal of Statistical Software 2015; 67(1):1–48.
  17. Loffing J, Loffing-Cueni D, Valderrabano V, Klausli L, Hebert SC, Rossier BC. Distribution of transcellular calcium and sodium transport pathways along mouse distal nephron.. American Journal of Physiology-Renal Physiology 2001; 281(6):1021–7.
    pubmed: 11704552
  18. Boros S, Bindels RJM, Hoenderop JGJ. Active Ca2+ reabsorption in the connecting tubule.. Pflugers Archiv-European Journal of Physiology 2009; 458(1):99–109.
    doi: 10.1007/s00424-008-0602-6pubmed: 18989697google scholar: lookup
  19. Bonny O, Edwards A. Calcium reabsorption in the distal tubule: regulation by sodium, pH, and flow.. American Journal of Physiology-Renal Physiology 2013; 304(5):585–600.
    pubmed: 23152295
  20. Wilkens MR, Kunert-Keil C, Brinkmeier H, Schroder B. Expression of calcium channel TRPV6 in ovine epithelial tissue.. Vet J 2009; 182(2):294–300.
    doi: 10.1016/j.tvjl.2008.06.020pubmed: 18701326google scholar: lookup
  21. Wilkens MR, Richter J, Fraser DR, Liesegang A, Breves G, Schroeder B. In contrast to sheep, goats adapt to dietary calcium restriction by increasing intestinal absorption of calcium.. Comparative Biochemistry and Physiology a-Molecular & Integrative Physiology 2012; 163(3–4):396–406.
    pubmed: 22776717
  22. Wilkens MR, Breves G, Schroder B. A goat is not a sheep: physiological similarities and differences observed in two ruminant species facing a challenge of calcium homeostatic mechanisms.. Animal Production Science 2014; 54(9):1507–11.
  23. Herm G, Muscher-Banse AS, Breves G, Schroder B, Wilkens MR. Renal mechanisms of calcium homeostasis in sheep and goats.. J Anim Sci 2015; 93(4):1608–21.
    doi: 10.2527/jas.2014-8450pubmed: 26020183google scholar: lookup
  24. Hoenderop JGJ, Hartog A, Stuiver M, Doucet A, Willems P, Bindels RJM. Localization of the epithelial Ca2+ channel in rabbit kidney and intestine.. J Am Soc Nephrol 2000; 11(7):1171–8.
    pubmed: 10864572
  25. Rourke KM, Coe S, Kohn CW, Rosol TJ, Mendoza FJ, Toribio RE. Cloning, comparative sequence analysis and mRNA expression of calcium-transporting genes in horses.. Gen Comp Endocrinol 2010; 167(1):6–10.
    doi: 10.1016/j.ygcen.2010.02.022pubmed: 20226785google scholar: lookup
  26. Hwang I, Jung EM, Yang H, Choi KC, Jeung EB. Tissue-Specific Expression of the Calcium Transporter Genes TRPV5, TRPV6, NCX1, and PMCA1b in the Duodenum, Kidney and Heart of Equus caballus.. J Vet Med Sci 2011; 73(11):1437–44.
    pubmed: 21737966
  27. Sprekeler N, Mueller T, Kowalewski MP, Liesegang A, Boos A. Expression patterns of intestinal calcium transport factors and ex-vivo absorption of calcium in horses.. BMC Vet Res 2011; 7.
    pmc: PMC3221622pubmed: 22017756
  28. Palm C, Hartmann K, Weber K. Expression and Immunolocalization of Calcium Transport Proteins in the Canine Duodenum, Kidney, and Pancreas.. Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology 2010; 293(5):770–4.
    pubmed: 20186965
  29. Kim JA, Yang H, Hwang I, Jung EM, Choi KC, Jeung EB. Expression Patterns and Potential Action of the Calcium Transport Genes Trpv5, Trpv6, Ncx1 and Pmca1b in the Canine Duodenum, Kidney and Uterus.. In Vivo 2011; 25(5):773–80.
    pubmed: 21753133
  30. Magyar CE, White KE, Rojas R, Apodaca G, Friedman PA. Plasma membrane Ca2+-ATPase and NCX1 Na+/Ca2+ exchanger expression in distal convoluted tubule cells.. American Journal of Physiology-Renal Physiology 2002; 283(1):29–40.
    pubmed: 12060584
  31. Haussler MR, Whitfield GK, Kaneko I, Haussler CA, Hsieh D, Hsieh J-C. Molecular Mechanisms of Vitamin D Action.. Calcif Tissue Int 2013; 92(2):77–98.
    doi: 10.1007/s00223-012-9619-0pubmed: 22782502google scholar: lookup
  32. Jones G, Prosser DE, Kaufmann M. 25-Hydroxyvitamin D-24-hydroxylase (CYP24A1): Its important role in the degradation of vitamin D.. Arch Biochem Biophys 2012; 523(1):9–18.
    doi: 10.1016/j.abb.2011.11.003pubmed: 22100522google scholar: lookup
  33. Hoenderop JGJ, Bindels RJM. Calciotropic and magnesiotropic TRP channels.. Physiology 2008; 23(1):32–40.
    pubmed: 18268363
  34. Hoenderop JGJ, Voets T, Hoefs S, Weidema F, Prenen J, Nilius B. Homo- and heterotetrameric architecture of the epithelial Ca2+ channels TRPV5 and TRPV6.. EMBO J 2003; 22(4):776–85.
    pmc: PMC145440pubmed: 12574114
  35. Hoenderop JGJ, Vennekens R, Muller D, Prenen J, Droogmans G, Bindels RJM. Function and expression of the epithelial Ca2+ channel family: comparison of mammalian ECaC1 and 2.. Journal of Physiology-London 2001; 537(3):747–61.
    pmc: PMC2278984pubmed: 11744752
  36. Hoenderop JGJ, Nilius B, Bindels RJM. Calcium absorption across epithelia.. Physiol Rev 2005; 85(1):373–422.
    pubmed: 15618484
  37. Mensenkamp AR, Hoenderop JGJ, Bindels RJM. TRPV5, the gateway to Ca2+ homeostasis.. Handb Exp Pharmacol 2007; (179):207–20.
    pubmed: 17217059
  38. Schoeber JPH, Hoenderop JGJ, Bindels RJM. Concerted action of associated proteins in the regulation of TRPVS and TRPV6.. Biochem Soc Trans 2007; 35:115–9.
    pubmed: 17233615
  39. Palviainen M, Raekallio M, Rajamaki MM, Linden J, Vainio O. Kidney-derived proteins in urine as biomarkers of induced acute kidney injury in sheep.. Vet J 2012; 193(1):287–9.
    doi: 10.1016/j.tvjl.2011.10.004pubmed: 22088561google scholar: lookup
  40. Stockham SL, Scott MA. Fundamentals of veterinary clinical pathology.. 2nd ed. Stockham SL, Scott MA, editors. Ames, Iowa: Blackwell Pub., c2008, 2008; Pp. 415–494.
  41. Borke JL, Caride A, Verma AK, Penniston JT, Kumar R. Plasma membrane calcium pump and 28-kDa calcium binding protein in cells of rat kidney distal tubules.. Am J Physiol 1989; 257(5):842–9.
    pubmed: 2556040
  42. Costanzo LS, Windhager EE. Calcium and sodium transport by the distal convoluted tubule of the rat.. Am J Physiol 1978; 235(5):492–506.
    pubmed: 727266
  43. Strehler EE. Plasma membrane calcium ATPases: From generic Ca2+ sump pumps to versatile systems for fine-tuning cellular Ca2+.. Biochem Biophys Res Commun 2015; 460(1):26–33.
    doi: 10.1016/j.bbrc.2015.01.121pubmed: 25998731google scholar: lookup
  44. Lefebvre HP, Dossin O, Trumel C, Braun J-P. Fractional excretion tests: a critical review of methods and applications in domestic animals.. Vet Clin Pathol 2008; 37(1):4–20.
    doi: 10.1111/j.1939-165X.2008.00010.xpubmed: 18366540google scholar: lookup
  45. Maenpaa PH, Koskinen T, Koskinen E. Serum profiles of vitamin A, vitamin E and vitamin D in mares and foals during different season. J Anim Sci 1988; 66(6):1418–23.
    pubmed: 3397359
  46. Azarpeykan S, Dittmer KE, Gee EK, Marshall JC, Wallace J, Elder P. Influence of blanketing and season on vitamin D and parathyroid hormone, calcium, phosphorus, and magnesium concentrations in horses in New Zealand.. Domest Anim Endocrinol 2016; 56:75–84.
    doi: 10.1016/j.domaniend.2016.03.003pubmed: 27131337google scholar: lookup
  47. Breidenbach A, Schlumbohm C, Harmeyer J. Peculiarities of vitamin D and of the calcium and phosphate homeostatic system in horses.. Vet Res 1998; 29(2):173–86.
    pubmed: 9601149
  48. Azarpeykan S, Dittmer KE, Gee EK, Marshall JC, Elder P, Acke E. Circadian rhythm of calciotropic hormones, serum calcium, phosphorus and magnesium during the shortest and longest days of the year in horses in New Zealand.. J Anim Physiol Anim Nutr (Berl) .
    doi: 10.1111/jpn.12477pubmed: 26841283google scholar: lookup

Citations

This article has been cited 6 times.
  1. Alemi M, Ahmadi Sheikhsarmast S, Mohri M. Serum 25(OH) Vitamin D Concentrations in Horses: Effects of Age, Gender, Breed, Skin Colour and Season. Vet Med Sci 2025 Jan;11(1):e70092.
    doi: 10.1002/vms3.70092pubmed: 39778002google scholar: lookup
  2. Dosi MCM, Riggs CM, May J, Lee A, Cillan-Garcia E, Pagan J, McGorum BC. Thoroughbred Racehorses in Hong Kong Require Vitamin D Supplementation to Mitigate the Risk of Low Vitamin D Status. Animals (Basel) 2023 Jun 29;13(13).
    doi: 10.3390/ani13132145pubmed: 37443942google scholar: lookup
  3. Khattar V, Wang L, Peng JB. Calcium selective channel TRPV6: Structure, function, and implications in health and disease. Gene 2022 Apr 5;817:146192.
    doi: 10.1016/j.gene.2022.146192pubmed: 35031425google scholar: lookup
  4. Dittmer KE, Chernyavtseva A, Marshall JC, Cabrera D, Wolber FM, Kruger M. Expression of Renal Vitamin D and Phosphatonin-Related Genes in a Sheep Model of Osteoporosis. Animals (Basel) 2021 Dec 29;12(1).
    doi: 10.3390/ani12010067pubmed: 35011173google scholar: lookup
  5. Dittmer KE, Heathcott RW, Marshall JC, Azarpeykan S. Expression of Phosphatonin-Related Genes in Sheep, Dog and Horse Kidneys Using Quantitative Reverse Transcriptase PCR. Animals (Basel) 2020 Oct 5;10(10).
    doi: 10.3390/ani10101806pubmed: 33027890google scholar: lookup
  6. Hurst EA, Homer NZ, Mellanby RJ. Vitamin D Metabolism and Profiling in Veterinary Species. Metabolites 2020 Sep 15;10(9).
    doi: 10.3390/metabo10090371pubmed: 32942601google scholar: lookup
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