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

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

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