Evaluation of high–molecular weight adiponectin in horses

Anne A. Wooldridge Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.
Boshell Diabetes and Metabolic Diseases Research Program, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Anne A. Wooldridge in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Heather Gray Edwards Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Heather Gray Edwards in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Eric P. Plaisance Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University Auburn, AL 36849.
Boshell Diabetes and Metabolic Diseases Research Program, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Eric P. Plaisance in
Current site
Google Scholar
PubMed
Close
 PhD
,
Rory Applegate Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Rory Applegate in
Current site
Google Scholar
PubMed
Close
 DVM
,
Debra R. Taylor Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Debra R. Taylor in
Current site
Google Scholar
PubMed
Close
 DVM, MS
,
Jennifer Taintor Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Jennifer Taintor in
Current site
Google Scholar
PubMed
Close
 DVM, MS
,
Qiao Zhong Department of Clinical Sciences, College of Veterinary Medicine, Auburn University Auburn, AL 36849.
Boshell Diabetes and Metabolic Diseases Research Program, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Qiao Zhong in
Current site
Google Scholar
PubMed
Close
, and
Robert L. Judd Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University Auburn, AL 36849.
Boshell Diabetes and Metabolic Diseases Research Program, College of Veterinary Medicine, Auburn University Auburn, AL 36849.

Search for other papers by Robert L. Judd in
Current site
Google Scholar
PubMed
Close
 PhD

Abstract

Objective—To characterize adiponectin protein complexes in lean and obese horses.

Animals—26 lean horses and 18 obese horses.

Procedures—Body condition score (BCS) and serum insulin activity were measured for each horse. Denaturing and native western blot analyses were used to evaluate adiponectin complexes in serum. A human ELISA kit was validated and used to quantify high–molecular weight (HMW) complexes. Correlations between variables were made, and HMW values were compared between groups.

Results—Adiponectin was present as a multimer consisting of HMW (> 720-kDa), low-molecular weight (180-kDa), and trimeric (90-kDa) complexes in serum. All complexes were qualitatively reduced in obese horses versus lean horses, but the percentage of complexes < 250 kDa was higher in obese versus lean horses. High–molecular weight adiponectin concentration measured via ELISA was negatively correlated with serum insulin activity and BCS and was lower in obese horses (mean ± SD, 3.6 ± 3.9 μg/mL), compared with lean horses (8.0 ± 4.6 μg/mL).

Conclusions and Clinical Relevance—HMW adiponectin is measurable via ELISA, and concentration is negatively correlated with BCS and serum insulin activity in horses. A greater understanding of the role of adiponectin in equine metabolism will provide insight into the pathophysiology of metabolic disease conditions.

Abstract

Objective—To characterize adiponectin protein complexes in lean and obese horses.

Animals—26 lean horses and 18 obese horses.

Procedures—Body condition score (BCS) and serum insulin activity were measured for each horse. Denaturing and native western blot analyses were used to evaluate adiponectin complexes in serum. A human ELISA kit was validated and used to quantify high–molecular weight (HMW) complexes. Correlations between variables were made, and HMW values were compared between groups.

Results—Adiponectin was present as a multimer consisting of HMW (> 720-kDa), low-molecular weight (180-kDa), and trimeric (90-kDa) complexes in serum. All complexes were qualitatively reduced in obese horses versus lean horses, but the percentage of complexes < 250 kDa was higher in obese versus lean horses. High–molecular weight adiponectin concentration measured via ELISA was negatively correlated with serum insulin activity and BCS and was lower in obese horses (mean ± SD, 3.6 ± 3.9 μg/mL), compared with lean horses (8.0 ± 4.6 μg/mL).

Conclusions and Clinical Relevance—HMW adiponectin is measurable via ELISA, and concentration is negatively correlated with BCS and serum insulin activity in horses. A greater understanding of the role of adiponectin in equine metabolism will provide insight into the pathophysiology of metabolic disease conditions.

Contributor Notes

Dr. Plaisance's present address is Laboratory of Nutrient Sensing and Adipocyte Signaling, Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, LA 70808.

Funding provided from Animal Health and Disease Research Funds, Auburn University.

Presented at the American College of Veterinary Internal Medicine Forum, Anaheim, Calif, June 2010; Boshell Diabetes and Metabolic Diseases Research Day, Auburn, Ala, March 2009; and Boshell Diabetes and Metabolic Diseases Research Day, Auburn, Ala, March 2010.

Address correspondence to Dr. Wooldridge (aaw0002@auburn.edu).
  • 1 Frank N, Geor RJ, Bailey SR, et al. Equine metabolic syndrome. J Vet Intern Med 2010; 24: 467475.

  • 2 Gordon ME, McKeever KH, Betros CL, et al. Plasma leptin, ghrelin and adiponectin concentrations in young fit racehorses versus mature unfit Standardbreds. Vet J 2007; 173: 91100.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 3 Gordon ME, McKeever KH, Betros CL, et al. Exercise-induced alterations in plasma concentrations of ghrelin, adiponectin, leptin, glucose, insulin, and cortisol in horses. Vet J 2007; 173: 532540.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4 Kearns CF, McKeever KH, Roegner V, et al. Adiponectin and leptin are related to fat mass in horses. Vet J 2006; 172: 460465.

  • 5 Gil-Campos M, Canete RR, Gil A. Adiponectin, the missing link in insulin resistance and obesity. Clin Nutr 2004; 23: 963974.

  • 6 Richards AA, Stephens T, Charlton HK, et al. Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: evidence for regulation of multimerization by alterations in posttranslational modifications. Mol Endocrinol 2006; 20: 16731687.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7 Schraw T, Wang ZV, Halberg N, et al. Plasma adiponectin complexes have distinct biochemical characteristics. Endocrinology 2008; 149: 22702282.

  • 8 Pajvani UB, Hawkins M, Combs TP, et al. Complex distribution, not absolute amount of adiponectin, correlates with thiazolidinedione-mediated improvement in insulin sensitivity. J Biol Chem 2004; 279: 1215212162.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9 Hug C, Wang J, Ahmad NS, et al. T-cadherin is a receptor for hexameric and high-molecular-weight forms of Acrp30/adiponectin. Proc Natl Acad Sci US A 2004; 101: 1030810313.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10 Radjainia M, Wang Y, Mitra AK. Structural polymorphism of oligomeric adiponectin visualized by electron microscopy. J Mol Biol 2008; 381: 419430.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11 Waki H, Yamauchi T, Kamon J, et al. Impaired multimerization of human adiponectin mutants associated with diabetes. Molecular structure and multimer formation of adiponectin. J Biol Chem 2003; 278: 4035240363.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12 Araki S, Dobashi K, Kubo K, et al. High molecular weight, rather than total, adiponectin levels better reflect metabolic abnormalities associated with childhood obesity. J Clin Endocrinol Metab 2006; 91: 51135116.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13 Wang AY, Hickman IJ, Richards AA, et al. High molecular weight adiponectin correlates with insulin sensitivity in patients with hepatitis C genotype 3, but not genotype 1 infection. Am J Gastroenterol 2005; 100: 27172723.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14 Almeda-Valdes P, Cuevas-Ramos D, Mehta R, et al. Total and high molecular weight adiponectin have similar utility for the identification of insulin resistance. Cardiovasc Diabetol 2010; 9: 26.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15 Liu Y, Retnakaran R, Hanley A, et al. Total and high molecular weight but not trimeric or hexameric forms of adiponectin correlate with markers of the metabolic syndrome and liver injury in Thai subjects. J Clin Endocrinol Metab 2007; 92: 43134318.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16 Bobbert T, Rochlitz H, Wegewitz U, et al. Changes of adiponectin oligomer composition by moderate weight reduction. Diabetes 2005; 54: 27122719.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17 Hara K, Horikoshi M, Yamauchi T, et al. Measurement of the high-molecular weight form of adiponectin in plasma is useful for the prediction of insulin resistance and metabolic syndrome. Diabetes Care 2006; 29: 13571362.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18 Inoue T, Kotooka N, Morooka T, et al. High molecular weight adiponectin as a predictor of long-term clinical outcome in patients with coronary artery disease. Am J Cardiol 2007; 100: 569574.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19 Kobayashi H, Ouchi N, Kihara S, et al. Selective suppression of endothelial cell apoptosis by the high molecular weight form of adiponectin. Circ Res 2004; 94: e27e31.

    • Search Google Scholar
    • Export Citation
  • 20 Salani B, Briatore L, Andraghetti G, et al. High-molecular weight adiponectin isoforms increase after biliopancreatic diversion in obese subjects. Obesity (Silver Spring) 2006; 14: 15111514.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 21 Swarbrick MM, Austrheim-Smith IT, Stanhope KL, et al. Circulating concentrations of high-molecular-weight adiponectin are increased following Roux-en-Y gastric bypass surgery. Diabetologia 2006; 49: 25522558.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 22 Torigoe M, Matsui H, Ogawa Y, et al. Impact of the high-molecular-weight form of adiponectin on endothelial function in healthy young men. Clin Endocrinol (Oxf) 2007; 67: 276281.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23 Wang ZV, Schraw TD, Kim JY, et al. Secretion of the adipocyte-specific secretory protein adiponectin critically depends on thiol-mediated protein retention. Mol Cell Biol 2007; 27: 37163731.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24 Aso Y, Yamamoto R, Wakabayashi S, et al. Comparison of serum high-molecular weight (HMW) adiponectin with total adiponectin concentrations in type 2 diabetic patients with coronary artery disease using a novel enzyme-linked immunosorbent assay to detect HMW adiponectin. Diabetes 2006; 55: 19541960.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25 Basu R, Pajvani UB, Rizza RA, et al. Selective downregulation of the high molecular weight form of adiponectin in hyperinsulinemia and in type 2 diabetes: differential regulation from nondiabetic subjects. Diabetes 2007; 56: 21742177.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26 Halperin F, Beckman JA, Patti ME, et al. The role of total and high-molecular-weight complex of adiponectin in vascular function in offspring whose parents both had type 2 diabetes. Diabetologia 2005; 48: 21472154.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27 Hara K, Horikoshi M, Yamauchi T, et al. Measurement of the high-molecular weight form of adiponectin in plasma is useful for the prediction of insulin resistance and metabolic syndrome. Diabetes Care 2006; 29: 13571362.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28 Lara-Castro C, Doud EC, Tapia PC, et al. Adiponectin multimers and metabolic syndrome traits: relative adiponectin resistance in African Americans. Obesity (Silver Spring) 2008; 16: 26162623.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29 Lara-Castro C, Luo N, Wallace P, et al. Adiponectin multimeric complexes and the metabolic syndrome trait cluster. Diabetes 2006; 55: 249259.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30 Murdolo G, Nowotny B, Celi F, et al. Inflammatory adipokines, high molecular weight adiponectin, and insulin resistance: a population-based survey in prepubertal schoolchildren. PLoS One 2011; 6:e17264.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31 Ebinuma H, Miyazaki O, Yago H, et al. A novel ELISA system for selective measurement of human adiponectin multimers by using proteases. Clin Chim Acta 2006; 372: 4753.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32 Nakano Y, Tajima S, Yoshimi A, et al. A novel enzyme-linked immunosorbent assay specific for high-molecular-weight adiponectin. J Lipid Res 2006; 47: 15721582.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 33 Sinha MK, Songer T, Xiao Q, et al. Analytical validation and biological evaluation of a high molecular-weight adiponectin ELISA. Clin Chem 2007; 53: 21442151.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 34 Gordon ME, McKeever KH. Diurnal variation of ghrelin, leptin, and adiponectin in Standardbred mares. J Anim Sci 2005; 83: 23652371.

  • 35 Gordon ME, McKeever KH. Oral and intravenous carbohydrate challenges decrease active ghrelin concentrations and alter hormones related to control of energy metabolism in horses. J Anim Sci 2006; 84: 16821690.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 36 Carter RA, Geor RJ, Burton Staniar W, et al. Apparent adiposity assessed by standardised scoring systems and morphometric measurements in horses and ponies. Vet J 2009; 179: 204210.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 37 Henneke DR, Potter GD, Kreider JL, et al. Relationship between condition score, physical measurements and body fat percentage in mares. Equine Vet J 1983; 15: 371372.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 38 Frank N. Equine metabolic syndrome. Vet Clin North Am Equine Pract 2011; 27: 7392.

  • 39 Freestone JF, Wolfsheimer KJ, Kamerling SG, et al. Exercise induced hormonal and metabolic changes in Thoroughbred horses: effects of conditioning and acepromazine. Equine Vet J 1991; 23: 219223.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 40 Wang Y, Lam KS, Yau MH, et al. Post-translational modifications of adiponectin: mechanisms and functional implications. Biochem J 2008; 409: 623633.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 41 Radin MJ, Sharkey LC, Holycross BJ. Adipokines: a review of biological and analytical principles and an update in dogs, cats, and horses. Vet Clin Pathol 2009; 38: 136156.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 42 Kusminski CM, Scherer PE. The road from discovery to clinic: adiponectin as a biomarker of metabolic status. Clin Pharmacol Ther 2009; 86: 592595.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 43 Murdolo G, Hammarstedt A, Schmelz M, et al. Acute hyperinsulinemia differentially regulates interstitial and circulating adiponectin oligomeric pattern in lean and insulin-resistant, obese individuals. J Clin Endocrinol Metab 2009; 94: 45084516.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 44 Bogan JS, Lodish HF. Two compartments for insulin-stimulated exocytosis in 3t3-L1 adipocytes defined by endogenous Acrp30 and Glut4. J Cell Biol 1999; 146: 609620.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 45 Shetty S, Kusminski CM, Scherer PE. Adiponectin in health and disease: evaluation of adiponectin-targeted drug development strategies. Trends Pharmacol Sci 2009; 30: 234239.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 46 Kubota N, Terauchi Y, Yamauchi T, et al. Disruption of adiponectin causes insulin resistance and neointimal formation. J Biol Chem 2002; 277: 2586325866.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 47 Maeda N, Shimomura I, Kishida K, et al. Diet-induced insulin resistance in mice lacking adiponectiNACRP30. Nat Med 2002; 8: 731737.

  • 48 Berg AH, Combs TP, Scherer PE. ACRP30/adiponectin: an adipokine regulating glucose and lipid metabolism. Trends Endocrinol Metab 2002; 13: 8489.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 49 Wang Y, Lam KS, Chan L, et al. Post-translational modifications of the four conserved lysine residues within the collagenous domain of adiponectin are required for the formation of its high molecular weight oligomeric complex. J Biol Chem 2006; 281: 1639116400.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 50 Wang Y, Xu A, Knight C, et al. Hydroxylation and glycosylation of the four conserved lysine residues in the collagenous domain of adiponectin. Potential role in the modulation of its insulin-sensitizing activity. J Biol Chem 2002; 277: 1952119529.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 51 Empana JP. Adiponectin isoforms and cardiovascular disease: the epidemiological evidence has just begun. Eur Heart J 2008; 29: 12211223.

    • Search Google Scholar
    • Export Citation
  • 52 Leth H, Andersen KK, Frystyk J, et al. Elevated levels of high-molecular-weight adiponectin in type 1 diabetes. J Clin Endocrinol Metab 2008; 93: 31863191.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 53 Cook JR, Semple RK. Hypoadiponectinemia—cause or consequence of human “insulin resistance”? J Clin Endocrinol Metab 2010; 95: 15441554.

  • 54 Pajvani UB, Du X, Combs TP, et al. Structure-function studies of the adipocyte-secreted hormone Acrp30/adiponectin. Implications for metabolic regulation and bioactivity. J Biol Chem 2003; 278: 90739085.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 55 Sun L, Hu FB, Yu Z, et al. Lean body mass, interleukin 18, and metabolic syndrome in apparently healthy Chinese. PLoS One 2011; 6:e18104.

  • 56 Yu D, Yu Z, Sun Q, et al. Effects of body fat on the associations of high-molecular-weight adiponectin, leptin and soluble leptin receptor with metabolic syndrome in Chinese. PLoS One 2011; 6:e16818.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 57 Combs TP, Berg AH, Rajala MW, et al. Sexual differentiation, pregnancy, calorie restriction, and aging affect the adipocyte-specific secretory protein adiponectin. Diabetes 2003; 52: 268276.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 58 Xu A, Chan KW, Hoo RL, et al. Testosterone selectively reduces the high molecular weight form of adiponectin by inhibiting its secretion from adipocytes. J Biol Chem 2005; 280: 1807318080.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 59 Wagner JA, Wright EC, Ennis MM, et al. Utility of adiponectin as a biomarker predictive of glycemic efficacy is demonstrated by collaborative pooling of data from clinical trials conducted by multiple sponsors. Clin Pharmacol Ther 2009; 86: 619625.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 60 Nawrocki AR, Scherer PE. Keynote review: the adipocyte as a drug discovery target. Drug Discov Today 2005; 10: 12191230.

Advertisement