Download PDF
Horm Metab Res 2007; 39(8): 548-554
DOI: 10.1055/s-2007-985142
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Quantitative Real-time PCR for the Measurement of 11β-HSD1 and 11β-HSD2 mRNA Levels in Tissues of Healthy Dogs

N. S. Sieber-Ruckstuhl 1 , M. L. Meli 2 , F. S. Boretti 1 , E. Gönczi 2 , H. Lutz 2 , C. E. Reusch 1
  • 1Clinic for Small Animal Internal Medicine, Vetsuisse Faculty University of Zurich, Zurich, Switzerland
  • 2Clinical Laboratory, Vetsuisse Faculty University of Zurich, Zurich, Switzerland
Further Information

Publication History

received 09.01.2007

accepted 05.03.2007

Publication Date:
21 August 2007 (online)

Also available at
    Permissions and Reprints

Abstract

The 11β-hydroxysteroid dehydrogenase (11β-HSD) exists in two isoforms, 11β-HSD1 and 11β-HSD2. 11β-HSD1 generates active cortisol from cortisone and appears to be involved in insulin resistant states. 11β-HSD2 protects the mineralocorticoid receptor from inappropriate activation by glucocorticoids and is important to prevent sodium retention and hypertension. The purposes of the present study were to develop two real-time PCR assays to assess 11β-HSD1 and 11β-HSD2 mRNA expression and to evaluate the tissue distribution of the two isoforms in dogs. Thirteen different tissues of 10 healthy dogs were evaluated. Both real-time PCR assays were highly specific, sensitive and reproducible. Highest 11β-HSD1 mRNA expression was seen in liver, lung, and renal medulla; highest 11β-HSD2 mRNA expression in renal cortex, adrenal gland, and renal medulla. Higher 11β-HSD1 than 11β-HSD2 mRNA levels were found in all tissues except adrenal gland, colon, and rectum. Our results demonstrate that the basic tissue distribution of 11β-HSD1 and 11β-HSD2 in dogs corresponds to that in humans and rodents. In a next step 11β-HSD1 and 11β-HSD2 expression should be assessed in diseases like obesity, hypercortisolism, and hypertension to improve our knowledge about 11β-HSD activity, to evaluate the dog as a model for humans and to potentially find new therapeutic options.

Key words

Real-time PCR - 11β-HSD1 - 11β-HSD2 - tissue distribution - dogs

References

  • 1 Krozowski Z, Li KXZ, Koyama K, Smith RE, Obeyesekere VR, Stein-Oakley A, Sasano H, Coulter C, Cole T, Sheppard KE. The type I and type II 11β-hydroxysteroid dehydrogenase enzymes.  J Steroid Biochem Mel Biol. 1999;  69 391-401
    PubMedGoogle Scholar
  • 2 Stewart PM, Krozowski ZS. 11β-hydroxysteroid dehydrogenase.  Vitam Horm. 1999;  57 249-324
    PubMedGoogle Scholar
  • 3 Agarwal AK, Monder C, Eckstein B, White PC. Cloning and expression of rat cDNA encoding coticosteroid 11β-dehydrogenase.  J Biol Chem. 1989;  264 18939-18943
    PubMedGoogle Scholar
  • 4 Ricketts ML, Verhaeg JM, Bujalska I, Howie AJ, Rainey WE, Stewart PM. Immunohistochemical localization of type 1 11β-hydroxysteroid dehydrogenase in human tissues.  J Clin Endocrinol Metab. 1998;  83 1325-1335
    CrossrefPubMedGoogle Scholar
  • 5 Albiston AL, Obeyesekere VR, Smith RE, Krozowski ZS. Cloning and tissue distribution of the human 11β-hydroxysteroid dehydrogenase type 2 enzyme.  Mol Cell Endocrinol. 1994;  105 R11-R17
    CrossrefPubMedGoogle Scholar
  • 6 Edwards CRW, Stewart PM, Burt D, Brett L, MacIntyre MA, Sutanto WE, Kloet ER De, Monder C. Localisation of 11β-hydroxysteroid dehydrogenase-tissue specific protector of the mineralocorticoid receptor.  Lancet. 1988;  2 986-989
    PubMedGoogle Scholar
  • 7 Tomlinson JW, Draper N, Mackie J, Johnson AP, Holder G, Wood P, Stewart PM. Absence of cushinoid phenotype in a patient with Cushing's disease due to defective cortisone to cortisol conversion.  J Clin Endocrinol Metab. 2002;  87 57-62
    CrossrefPubMedGoogle Scholar
  • 8 Paulmyer-Lacroix O, Boullu S, Oliver C, Alessi M, Grino M. Expression of the mRNA coding for 11β-hydroxysteroid dehydrogenase type 1 in adipose tissue from obese patients: An in situ hybridization study.  J Clin Endocrinol Metab. 2002;  87 2701-2705
    CrossrefPubMedGoogle Scholar
  • 9 Wake DJ, Rask E, Livingstone DEW, Söderberg S, Olsson T, Walker BR. Local and systemic impact of transcriptional up-regulation of 11β-hydroxysteroid dehydrogenase type 1 in adipose tissue in human obesity.  J Clin Endocrinol Metab. 2003;  88 3983-3988
    CrossrefPubMedGoogle Scholar
  • 10 Valsamakis G, Anwar A, Tomlinson JW, Shackleton CHL, MacTernan PG, Chetty R, Wood PJ, Banerjee AK, Holder G, Barnett AH, Stewart PM, Kumar S. 11β-hydroxysteroid dehydrogenase type 1 activity in lean and obese males with type 2 diabetes mellitus.  J Clin Endocrinol Metab. 2004;  89 4755-4761
    CrossrefPubMedGoogle Scholar
  • 11 Desbriere R, Vuaroqueaux V, Achard V, Boullu-Ciocca S, Labuhn M, Dutour A, Grino M. 11β-hydroxysteroid dehydrogenase type 1 mRNA is increased in both visceral and subcutaneous adipose tissue of obese patients.  Obesity. 2006;  14 794-798
    CrossrefPubMedGoogle Scholar
  • 12 Alberts P, Rönquist-Nii Y, Larsson C, Klingström G, Engblom L, Edling N, Lidell V, Berg I, Edlund PO, Ashkzari M, Sahaf N, Norling S, Berggren V, Bergdahl K, Forsgren M, Abrahmsén L. Effect of high-fat diet on KKAy and ob/ob mouse liver and adipose tissue corticosterone and 11-dehydrocorticosterone concentrations.  Horm Metab Res. 2005;  37 402-407
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Masuzaki H, Flier JS. Tissue-specific glucocorticoid reactivating enzyme, 11 beta-hydroxysteroid dehydrogenase type 1 (11 beta-HSD1)-a promising drug target for the treatment of metabolic syndrome.  Curr Drug Targets Immune Endocr Metabol Disord. 2003;  3 255-262
    CrossrefPubMedGoogle Scholar
  • 14 Chrousos GP. Is 11β-hydroxysteroid dehydrogenase type 1 a good therapeutic target for blockade of glucocorticoid actions?.  Proc Natl Acad Sci USA. 2004;  101 6329-6330
    CrossrefPubMedGoogle Scholar
  • 15 Li A, Li KXZ, Marui S, Krozowski ZS, Batista MC, Whorwood CB, Arnold IJP, Shackleton CHL, Mendonca BB, Stewart PM. Apparent mineralocorticoid excess in a Brazilian kindred: hypertension in the heterozygote state.  J Hypertens. 1997;  15 1397-1402
    CrossrefPubMedGoogle Scholar
  • 16 Pretorius P, Wallner B, Marx J. Cortisol resistance in conditions such as asthma and the involvement of 11β-HSD-2: A hypothesis.  Horm Metab Res. 2006;  38 368-376
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Arampatzis S, Kadereit B, Schuster D, Balazs Z, Schweizer RAS, Frey FJ, Langer T, Odermatt A. Comparative enzymology of 11β-hydroxysteroid dehydrogenase type 1 from six species.  J Mol Endocrinol. 2005;  35 89-101
    CrossrefPubMedGoogle Scholar
  • 18 Slight S, Ganjam VK, Weber KT. Species diversity of 11beta-hydroxysteroid dehydrogenase in the cardiovascular system.  J Lab Clin Med. 1994;  124 821-826
    PubMedGoogle Scholar
  • 19 Whorwood CB, Franklyn JA, Sheppard MC, Stewart PM. Tissue localization of 11 beta-hydroxysteroid dehydrogenase and its relationship to the glucocorticoid receptor.  J Steroid Biochem Mol Biol. 1992;  41 21-28
    CrossrefPubMedGoogle Scholar
  • 20 Klein D, Janda P, Steinborn R, Muller M, Salmons B, Gunzburg WH. Proviral load determination of different feline immunodeficiency virus isolates using real-time polymerase chain reaction: influence of mismatches on quantification.  Electrophoresis. 1999;  20 291-299
    CrossrefPubMedGoogle Scholar
  • 21 McTernan CL, Draper N, Nicholson H, Chalder SM, Driver P, Hewison M, Kilby MD, Stewart PM. Reduced placental 11β-hydroxysteroid dehydrogenase type 2 mRNA levels in human pregnancies complicated by intrauterine growth restriction: an analysis of possible mechanisms.  J Clin Endocrinol Metab. 2001;  86 4979-4983
    CrossrefPubMedGoogle Scholar
  • 22 Mazancova K, Miksik I, Kunes J, Pacha J. Placental 11β-hydroxysteroid dehydrogenase in dahl and spontaneously hypertensive rats.  Am J Hypertens. 2003;  16 401-406
    CrossrefPubMedGoogle Scholar
  • 23 Poletto R, Steibel JP, Siegford JM, Zanella AJ. Effects of early weaning and social isolation on the expression of glucocorticoid and mineralocorticoid receptor and 11β-hydroxysteroid dehydrogenase 1 and 2 mRNAs in the frontal cortex and hippocampus of piglets.  Brain Res. 2006;  1067 36-42
    CrossrefPubMedGoogle Scholar
  • 24 Rajan V, Chapman KE, Lyons V, Jamieson P, Mullins JJ, Edwards CRW, Seckl JR. Cloning, sequencing and tissue-distribution of mouse 11β-hydroxysteroid dehydrogenase-1 cDNA.  J Steroid Biochem Mol Biol. 1995;  52 141-147
    CrossrefPubMedGoogle Scholar
  • 25 Stewart PM, Murry BA, Mason JI. Human renal 11β-hydroxysteroid dehydrogenase is a high affinity nicotinamide adenine dinucleotide-dependent enzyme and differs from the cloned type I isoform.  J Clin Endocrinol Metab. 1994;  79 480-484
    CrossrefPubMedGoogle Scholar
  • 26 Shimojo M, Whorwood CB, Stewart PM. 11beta-hydroxysteroid dehydrogenase in the rat adrenal.  J Mol Endocrinol. 1996;  17 112-130
    PubMedGoogle Scholar
  • 27 Lombès M, Alfaidy N, Eugene E, Lessana A, Farman N, Bonvalet J. Prerequisite for cardiac aldosterone action: mineralocorticoid receptor and 11β-hydroxysteroid dehydrogenase in the human heart.  Circulation. 1995;  92 175-182
    PubMedGoogle Scholar
  • 28 Slight SH, Ganjam VK, Gómez-Sánchez CE, Zhou M, Weber KT. High affinity NAD+-dependent 11β-hydroxysteroid dehydrogenase in the human heart.  J Mol Cell Cardiol. 1996;  28 781-787
    CrossrefPubMedGoogle Scholar
  • 29 Albiston AL, Smith RE, Krozowski ZS. Sex- and tissue-specific regulation of 11β-hydroxysteroid dehydrogenase mRNA.  Mol Cell Endocrinol. 1995;  109 183-188
    CrossrefPubMedGoogle Scholar
  • 30 Raven PW, Taylor NF. Sex differences in the human metabolism of cortisol.  Endocr Res. 1996;  2284 751-755
    PubMedGoogle Scholar

Correspondence

N. S. Sieber-Ruckstuhl

Clinic for Small Animal Internal Medicine

Vetsuisse Faculty University of Zurich

Winterthurerstrasse 260

8057 Zurich

Switzerland

Phone: +41/44/635 83 01

Fax: +41/44/635 89 30

Email: nsieber@vetclinics.unizh.ch

      >