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Home / Equine Research Bank / J Vet Med Sci / Sensitive radioimmunoassay of total thyroxine (T4) in horses...
The Journal of veterinary medical science2017; 79(7); 1294-1300; doi: 10.1292/jvms.17-0133

Sensitive radioimmunoassay of total thyroxine (T4) in horses using a simple extraction method.

Abstract: Most thyroid hormone determinations in animals are based on immunoassays adapted from those used to test human samples, which may not reflect the actual values of thyroid hormone in horses because of the presence of binding proteins. The aims of the present study were i) to establish a novel radioimmunoassay (RIA) using a more simple and convenient method to separate binding proteins for the measurement of total thyroxine (T4) in horses and ii) to validate the assay by comparing total T4 concentrations in yearling horses raised in different climates. Blood samples were collected from trained yearlings in Hokkaido (temperate climate) and Miyazaki (subtropical climate) in Japan and from adult horses in estrus and diestrus. T4 was extracted from both serum and plasma using modified acid ethanol cryo-precipitation and sodium acetate ethanol methods. Circulating total T4 concentrations were determined by RIA. T4 concentration by sodium acetate ethanol was appropriately detectable rather than sodium salicylate method and was the same as for acid ethanol method. Furthermore, this sodium acetate ethanol method required fewer extraction steps than the other methods. Circulating T4 concentrations in yearlings were 225.98 ± 20.89 ng/ml, which was higher than the previous reference values. With respect to climate, T4 levels in Hokkaido yearlings tended to be higher than those in Miyazaki yearlings throughout the study period. These results indicated that this RIA protocol using a modified sodium acetate ethanol separation technique might be an appropriate tool for specific measurement of total T4 in horses.
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Publication Date: 2017-07-11 PubMed ID: 28603213PubMed Central: PMC5559379DOI: 10.1292/jvms.17-0133Google 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 presents a study that develops and validates a simpler, more convenient method for measuring total thyroxine (T4), a thyroid hormone, in horses. Their method is a novel radioimmunoassay (RIA) that avoids problems with existing techniques focused on human samples that don’t account for binding proteins in animals. It also compares hormone concentrations in horses from different climatic conditions.

Research Objectives

  • The primary aim of this study was to establish a new radioimmunoassay (RIA) that uses a simpler, more accessible method to separate binding proteins when measuring total thyroxine (T4) levels in horses.
  • The second aim was to validate this new assay by comparing the total T4 concentrations observed in yearling horses raised in different climates.

Methodology

  • Blood samples were collected from trained yearlings in Hokkaido and Miyazaki in Japan, two areas with varying climates: temperate and subtropical, respectively. Samples were also taken from adult horses in estrus and diestrus.
  • The researchers separated T4 from both serum and plasma using modified acid ethanol cryo-precipitation and sodium acetate ethanol methods.
  • Circulating total T4 concentrations were then determined by the newly developed RIA.

Findings

  • T4 concentration was more detectable using the sodium acetate ethanol method compared to other methods. Additionally, this method required fewer extraction steps.
  • The study found that circulating T4 concentrations in yearlings were higher than previous reference values.
  • The researchers observed that T4 levels in Hokkaido yearlings were higher overall than those from Miyazaki yearlings throughout the duration of the study.

Conclusion

  • Based on their results, the researchers suggested that the RIA protocol they employed, featuring a modified sodium acetate ethanol separation technique, could be an appropriate tool for specifically measuring total T4 levels in horses.

Cite This Article

APA
Tangyuenyong S, Nambo Y, Nagaoka K, Tanaka T, Watanabe G. (2017). Sensitive radioimmunoassay of total thyroxine (T4) in horses using a simple extraction method. J Vet Med Sci, 79(7), 1294-1300. https://doi.org/10.1292/jvms.17-0133

Publication

The Journal of veterinary medical science
ISSN: 1347-7439
NlmUniqueID: 9105360
Country: Japan
Language: English
Volume: 79
Issue: 7
Pages: 1294-1300

Researcher Affiliations

Tangyuenyong, Siriwan
  • Department of Basic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, Gifu 501-1193, Japan.
  • Laboratory of Veterinary Physiology, Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Nambo, Yasuo
  • Department of Basic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, Gifu 501-1193, Japan.
  • Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-0834, Japan.
Nagaoka, Kentaro
  • Department of Basic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, Gifu 501-1193, Japan.
  • Laboratory of Veterinary Physiology, Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Tanaka, Tomomi
  • Department of Basic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, Gifu 501-1193, Japan.
  • Laboratory of Veterinary Reproduction, Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Watanabe, Gen
  • Department of Basic Veterinary Science, United Graduate School of Veterinary Sciences, Gifu University, Gifu 501-1193, Japan.
  • Laboratory of Veterinary Physiology, Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.

MeSH Terms

  • Animals
  • Diestrus / blood
  • Estrus / blood
  • Female
  • Horses / blood
  • Male
  • Radioimmunoassay / methods
  • Radioimmunoassay / veterinary
  • Thyroxine / blood

References

This article includes 39 references
  1. Bird JA, Clarke L, Symonds ME. Influence of thyrotrophin-releasing hormone on thermoregulation in newborn lambs.. Biol Neonate 1998;73(1):52-9.
    doi: 10.1159/000013960pubmed: 9458943google scholar: lookup
  2. Breuhaus BA, Refsal KR, Beyerlein SL. Measurement of free thyroxine concentration in horses by equilibrium dialysis.. J Vet Intern Med 2006 Mar-Apr;20(2):371-6.
    doi: 10.1111/j.1939-1676.2006.tb02870.xpubmed: 16594596google scholar: lookup
  3. Brinkmann L, Gerken M, Hambly C, Speakman JR, Riek A. Thyroid hormones correlate with field metabolic rate in ponies, Equus ferus caballus.. J Exp Biol 2016 Aug 15;219(Pt 16):2559-66.
    doi: 10.1242/jeb.138784pubmed: 27312472google scholar: lookup
  4. Chen CL, Riley AM. Serum thyroxine and triiodothyronine concentrations in neonatal foals and mature horses.. Am J Vet Res 1981 Aug;42(8):1415-7.
    pubmed: 7294478
  5. Chopra IJ, Sack J, Fisher DA. 3,3',5'-Triiodothyronine (reverse T3) and 3,3',5-triiodothyronine (T3) in fetal and adult sheep: studies of metabolic clearance rates, production rates, serum binding, and thyroidal content relative to thyroxine.. Endocrinology 1975 Nov;97(5):1080-8.
    doi: 10.1210/endo-97-5-1080pubmed: 171141google scholar: lookup
  6. Daughaday WH, Mariz IK, Blethen SL. Inhibition of access of bound somatomedin to membrane receptor and immunobinding sites: a comparison of radioreceptor and radioimmunoassay of somatomedin in native and acid-ethanol-extracted serum.. J Clin Endocrinol Metab 1980 Oct;51(4):781-8.
    doi: 10.1210/jcem-51-4-781pubmed: 6998997google scholar: lookup
  7. Dudley RA, Edwards P, Ekins RP, Finney DJ, McKenzie IG, Raab GM, Rodbard D, Rodgers RP. Guidelines for immunoassay data processing.. Clin Chem 1985 Aug;31(8):1264-71.
    pubmed: 3893796
  8. Elliott KH, Welcker J, Gaston AJ, Hatch SA, Palace V, Hare JF, Speakman JR, Anderson WG. Thyroid hormones correlate with resting metabolic rate, not daily energy expenditure, in two charadriiform seabirds.. Biol Open 2013 Jun 15;2(6):580-6.
    doi: 10.1242/bio.20134358pmc: PMC3683160pubmed: 23789108google scholar: lookup
  9. Engelking L. R.2002. Review of Veterinary Physiology, Teton NewMedia, Wyoming.
  10. Eshratkhah B, Rajabian H, Namvar D, Eshratkhah S, Bastam SM. Comparative study on determination of plasma thyroid hormones by chemiluminescence and electrochemiluminescence immunoassay methods in sheep. Comp. Clin. Pathol. 2011;20:135–138.
    doi: 10.1007/s00580-010-0967-8google scholar: lookup
  11. Esquivel MS, Ramírez LC. Measurement of thyroid hormones and cortisol in horses with an automated immunoassay analyzer. Rev. Ciencias Veterinarias 2016;34:39–49.
  12. Evinger JV, Nelson RW. The clinical pharmacology of thyroid hormones in the dog.. J Am Vet Med Assoc 1984 Aug 1;185(3):314-6.
    pubmed: 6469837
  13. Fazio E, Medica P, Cravana C, Ferlazzo A. Total and free iodothyronines profile in the donkey (Equus asinus) over a 12-month period. Acta Vet. Brno 2012;81:239–244.
    doi: 10.2754/avb201281030239google scholar: lookup
  14. Fitzgerald BP, Davison LA. Thyroxine concentrations are elevated in mares which continue to exhibit estrous cycles during the nonbreeding season. JEVS 1998;18:48–51.
  15. Graves EA, Schott HC 2nd, Marteniuk JV, Refsal KR, Nachreiner RF. Thyroid hormone responses to endurance exercise.. Equine Vet J Suppl 2006 Aug;(36):32-6.
    doi: 10.1111/j.2042-3306.2006.tb05509.xpubmed: 17402388google scholar: lookup
  16. Gueorguieva R, Krystal JH. Move over ANOVA: progress in analyzing repeated-measures data and its reflection in papers published in the Archives of General Psychiatry.. Arch Gen Psychiatry 2004 Mar;61(3):310-7.
    doi: 10.1001/archpsyc.61.3.310pubmed: 14993119google scholar: lookup
  17. Gupta AK, Kumar S, Pal Y. Biochemical, haematological and thyroid hormone profile in healthy Indian Kathiawari horses. Asian-Aust. J. Anim. Sci. 2002;15:1215–1221.
  18. Hamada T, Watanabe G, Kokuho T, Taya K, Sasamoto S, Hasegawa Y, Miyamoto K, Igarashi M. Radioimmunoassay of inhibin in various mammals.. J Endocrinol 1989 Sep;122(3):697-704.
    doi: 10.1677/joe.0.1220697pubmed: 2509617google scholar: lookup
  19. Hasegawa Y, Miyamoto K, Fukuda M, Takahashi Y, Igarashi M. Immunological study of ovarian inhibin.. Endocrinol Jpn 1986 Oct;33(5):645-54.
    doi: 10.1507/endocrj1954.33.645pubmed: 3104019google scholar: lookup
  20. Huszenicza G, Nagy P, Juhász J, Kóródi P, Kulcsár M, Reiczigel J, Guillaume D, Rudas P, Solti L. Relationship between thyroid function and seasonal reproductive activity in mares.. J Reprod Fertil Suppl 2000;(56):163-72.
    pubmed: 20681128
  21. Kadunc NC, Kosec M, Cestnik V. Serum Triiodothyronine (T3) and Thyroxin (T4) concentrations in Lipizzan horses. Acta Vet. Brno 2003;72:17–22.
    doi: 10.2754/avb200372010017google scholar: lookup
  22. McArthur JN, Smith MJ. Binding of salicylate to albumin.. FEBS Lett 1968 Oct;1(5):332-334.
    doi: 10.1016/0014-5793(68)80148-6pubmed: 11945336google scholar: lookup
  23. Medica P, Fazio E, Cravana C, Ferlazzo A. Influence of endemic goitre areas on thyroid hormones in horses.. Animal 2011 Jan;5(1):82-7.
    doi: 10.1017/S175173111000145Xpubmed: 22440705google scholar: lookup
  24. Mejdell CM, Bøe KE. Responses to climatic variables of horses housed outdoors under Nordic winter conditions. Can. J. Anim. Sci. 2005;85:307–308.
    doi: 10.4141/A04-066google scholar: lookup
  25. Messer NT. Clinical and diagnostic features of thyroid disease in horses. Proc 11th ACVIM Forum, Washington, DC 1993:649–651.
  26. Nomura Y, Abe Y, Ishii Y, Watanabe M, Kobayashi M, Hattori A, Tsujimoto M. Structural changes in the glycosaminoglycan chain of rat skin decorin with growth.. J Dermatol 2003 Sep;30(9):655-64.
    doi: 10.1111/j.1346-8138.2003.tb00454.xpubmed: 14578555google scholar: lookup
  27. Reed S. M., Bayly W. M., Sellon D. C.2004. Equine Internal Medicine, 2nd ed., W.B. Saunders Co., St. Louis.
  28. Singh AK, Jiang Y, White T, Spassova D. Validation of nonradioactive chemiluminescent immunoassay methods for the analysis of thyroxine and cortisol in blood samples obtained from dogs, cats, and horses.. J Vet Diagn Invest 1997 Jul;9(3):261-8.
    doi: 10.1177/104063879700900307pubmed: 9249165google scholar: lookup
  29. Slebodziński AB, Brzezińska-Slebodzińska E, Nowak J, Kowalska K. Triiodothyronine (T3), insulin and characteristics of 5'-monodeiodinase (5'-MD) in mare's milk from parturition to 21 days post-partum.. Reprod Nutr Dev 1998 May-Jun;38(3):235-44.
    doi: 10.1051/rnd:19980303pubmed: 9698274google scholar: lookup
  30. Sojka JE, Johnson MA, Bottoms GD. Serum triiodothyronine, total thyroxine, and free thyroxine concentrations in horses.. Am J Vet Res 1993 Jan;54(1):52-5.
    pubmed: 8427472
  31. Sophianopoulos J, Jerkunica I, Lee CN, Sgoutas D. An improved ultrafiltration method for free thyroxine and triiodothyronine in serum.. Clin Chem 1980 Jan;26(1):159-62.
    pubmed: 7356554
  32. Sturman JA, Smith MJ. The binding of salicylate to plasma proteins in different species.. J Pharm Pharmacol 1967 Sep;19(9):621-3.
    doi: 10.1111/j.2042-7158.1967.tb09600.xpubmed: 4383147google scholar: lookup
  33. Todini L, Malfatti A, Salimei E, Fantuz F. Measurement of thyroid hormones in donkey (Equus asinus) blood and milk: validation of ELISA kits and evaluation of sample collection, handling and storage.. J Dairy Res 2010 Nov;77(4):419-24.
    doi: 10.1017/S0022029910000646pubmed: 20822570google scholar: lookup
  34. Todini L, Salimei E, Malfatti A, Brunetti VL, Fantuz F. Thyroid hormones in donkey blood and milk: Correlations with milk yield and environmental temperatures. Ital. J. Anim. Sci. 2015;14:4089.
    doi: 10.4081/ijas.2015.4089google scholar: lookup
  35. Tohei A, Akai M, Tomabechi T, Mamada M, Taya K. Adrenal and gonadal function in hypothyroid adult male rats.. J Endocrinol 1997 Jan;152(1):147-54.
    doi: 10.1677/joe.0.1520147pubmed: 9014850google scholar: lookup
  36. Ugrinowitsch C, Fellingham GW, Ricard MD. Limitations of ordinary least squares models in analyzing repeated measures data.. Med Sci Sports Exerc 2004 Dec;36(12):2144-8.
    doi: 10.1249/01.MSS.0000147580.40591.75pubmed: 15570152google scholar: lookup
  37. Wang R, Nelson JC, Wilcox RB. Salsalate and salicylate binding to and their displacement of thyroxine from thyroxine-binding globulin, transthyrin, and albumin.. Thyroid 1999 Apr;9(4):359-64.
    doi: 10.1089/thy.1999.9.359pubmed: 10319941google scholar: lookup
  38. Welcker J, Chastel O, Gabrielsen GW, Guillaumin J, Kitaysky AS, Speakman JR, Tremblay Y, Bech C. Thyroid hormones correlate with basal metabolic rate but not field metabolic rate in a wild bird species.. PLoS One 2013;8(2):e56229.
    doi: 10.1371/journal.pone.0056229pmc: PMC3577771pubmed: 23437096google scholar: lookup
  39. Wheeler MJ. A short history of hormone measurement.. Methods Mol Biol 2013;1065:1-6.
    doi: 10.1007/978-1-62703-616-0_1pubmed: 23996354google scholar: lookup

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