This Article Full Text Full Text (PDF) Submit a related Letter to the Editor Purchase Article View Shopping Cart Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hermansson, M. Articles by Carlsson, L. M. S. Search for Related Content PubMed PubMed Citation Articles by Hermansson, M. Articles by Carlsson, L. M. S.

Measurement of Human Growth Hormone Receptor Messenger Ribonucleic Acid by a Quantitative Polymerase Chain Reaction-Based Assay: Demonstration of Reduced Expression after Elective Surgery¹

Research Center for Endocrinology and Metabolism, Department of Internal Medicine, Sahlgrenska Hospital (M.H., R.B.W., R.B., B.C., L.M.S.C.), and the Department of Pediatrics, East Hospital (R.B.), Goteborg University, Goteborg; and the Department of Surgery, St. Goran Hospital (F.H., I.W.), and the Anesthesiological Metabolism Unit, Clinical Research Center, Department of Anesthesiology and Intensive Care, Huddinge University Hospital (J.W.), Karolinska Institute, Stockholm, Sweden

Address all correspondence and requests for reprints to: Dr. L. M. S. Carlsson, Research Center for Endocrinology and Metabolism, Department of Internal Medicine, Sahlgrenska Hospital, Goteborg University, S-413 45 Goteborg, Sweden. E-mail: lena{at}ss.gu.se

Studies of GH receptor (GHR) gene expression in human tissues have been hampered by the limited amount of tissue available for analysis and the low sensitivity of conventional methods. We have developed a quantitative reverse transcriptase-PCR assay for measurement of GHR messenger ribonucleic acid levels in small human tissue biopsies. To compensate for sample to sample variation, an internal RNA standard, which differs from the wild-type GHR transcript by only a few nucleotides, was reverse transcribed and amplified together with the GHR transcripts. PCR was carried out using one biotinylated primer to permit the purification of single stranded PCR products on streptavidin-coated microtiter plates. The ratio between the wild-type and mutated transcripts was determined by two separate minisequence reactions in which a primer, annealed immediately 3' of a variable nucleotide, was extended by a single ³H-labeled nucleotide, complementary to either the wild-type or mutated sequence. The assay range was 0.125–8 x 10⁵ transcripts/sample, the mean intraassay coefficient of variation was 8.7%, and the lower limit of detection was 0.125 x 10⁵ transcripts/sample. GHR messenger ribonucleic acid levels were detectable in small amounts (10–100 ng) of total RNA extracted from adipose tissue, skeletal muscle, and liver. The GHR gene expression in liver was approximately 10-fold higher than that in skeletal muscle, whereas intermediate levels were found in adipose tissue. In nine patients undergoing elective abdominal surgery, GHR gene expression in skeletal muscle was reduced on day 3 after surgery compared to the baseline level. The decrease in GHR gene expression was accompanied by a decrease in skeletal muscle glutamine. This suggests that the postoperative protein catabolism may be caused at least partly by acquired GH insensitivity due to reduced expression of the GHR gene.

This article has been cited by other articles:

				Y. Wei, Z. Rhani, and C. G. Goodyer Characterization of Growth Hormone Receptor Messenger Ribonucleic Acid Variants in Human Adipocytes J. Clin. Endocrinol. Metab., May 1, 2006; 91(5): 1901 - 1908. [Abstract] [Full Text] [PDF]

				D. Mesotten, P. J. Wouters, R. P. Peeters, K. V. Hardman, J. M. Holly, R. C. Baxter, and G. Van den Berghe Regulation of the Somatotropic Axis by Intensive Insulin Therapy during Protracted Critical Illness J. Clin. Endocrinol. Metab., July 1, 2004; 89(7): 3105 - 3113. [Abstract] [Full Text] [PDF]

				S. Fisker, K. Kristensen, A. M. Rosenfalck, S. B. Pedersen, L. Ebdrup, B. Richelsen, J. Hilsted, J. S. Christiansen, and J. O. L. Jørgensen Gene Expression of a Truncated and the Full-Length Growth Hormone (GH) Receptor in Subcutaneous Fat and Skeletal Muscle in GH-Deficient Adults: Impact of GH Treatment J. Clin. Endocrinol. Metab., February 1, 2001; 86(2): 792 - 796. [Abstract] [Full Text]

				Y. Zhang, J. Jiang, R. A. Black, G. Baumann, and S. J. Frank Tumor Necrosis Factor-{{alpha}} Converting Enzyme (TACE) Is a Growth Hormone Binding Protein (GHBP) Sheddase: The Metalloprotease TACE/ADAM-17 Is Critical for (PMA-Induced) GH Receptor Proteolysis and GHBP Generation Endocrinology, December 1, 2000; 141(12): 4342 - 4348. [Abstract] [Full Text] [PDF]

				M. Ballesteros, K.-C. Leung, R. J. M. Ross, T. P. Iismaa, and K. K. Y. Ho Distribution and Abundance of Messenger Ribonucleic Acid for Growth Hormone Receptor Isoforms in Human Tissues J. Clin. Endocrinol. Metab., August 1, 2000; 85(8): 2865 - 2871. [Abstract] [Full Text]

				K. J. Osterziel, R. Dietz, M. B. Ranke, G. Van den Berghe, J. Takala, and C. J. Hinds Increased Mortality Associated with Growth Hormone Treatment in Critically Ill Adults N. Engl. J. Med., January 13, 2000; 342(2): 134 - 136. [Full Text]

				G. Van den Berghe, R. C. Baxter, F. Weekers, P. Wouters, C. Y. Bowers, and J. D. Veldhuis A Paradoxical Gender Dissociation within the Growth Hormone/Insulin-Like Growth Factor I Axis during Protracted Critical Illness J. Clin. Endocrinol. Metab., January 1, 2000; 85(1): 183 - 192. [Abstract] [Full Text]

				Y. Mao, P.-R. Ling, T. P. Fitzgibbons, K. C. McCowen, G. P. Frick, B. R. Bistrian, and R. J. Smith Endotoxin-Induced Inhibition of Growth Hormone Receptor Signaling in Rat Liver in Vivo Endocrinology, December 1, 1999; 140(12): 5505 - 5515. [Abstract] [Full Text]

				G. Van den Berghe, F. de Zegher, and R. Bouillon Acute and Prolonged Critical Illness as Different Neuroendocrine Paradigms J. Clin. Endocrinol. Metab., June 1, 1998; 83(6): 1827 - 1834. [Full Text]

				R. Bjarnason, R. Wickelgren, M. Hermansson, F. Hammarqvist, B. Carlsson, and L. M. S. Carlsson Growth Hormone Treatment Prevents the Decrease in Insulin-Like Growth Factor I Gene Expression in Patients Undergoing Abdominal Surgery J. Clin. Endocrinol. Metab., May 1, 1998; 83(5): 1566 - 1572. [Abstract] [Full Text]

				Y. Zhang, R. Guan, J. Jiang, J. J. Kopchick, R. A. Black, G. Baumann, and S. J. Frank Growth Hormone (GH)-induced Dimerization Inhibits Phorbol Ester-stimulated GH Receptor Proteolysis J. Biol. Chem., June 29, 2001; 276(27): 24565 - 24573. [Abstract] [Full Text] [PDF]