Quantification of growth hormone receptor extra- and intracellular domain gene expression in chicken liver by quantitative competitive RT-PCR

General and Comparative Endocrinology
P Van AsE Decuypere

Abstract

The very sensitive competitive reverse transcription-polymerase chain reaction (RT-PCR) was used to investigate the expression of the extracellular (GHRe) and intracellular (GHRi) parts of the growth hormone receptor (GHR) in the liver tissue of chickens. Two competitors (internal standards), pGHRi MUT and pGHRe MUT, specific to the GHRi and GHRe genes, respectively, were constructed by site-specific mutagenesis. The internal standards defined PCR products of 394 bp for the pGHRi MUT and 330 bp for the GHRe MUT. These were used as competitors to the wild-type GHRi or GHRe which defined PCR products of 382 and 328 bp, respectively. Coamplification, under standardized conditions, of the native RNA in competition with serial dilutions of the mutant RNA in the same PCR reaction followed by enzymatic digestion produced the expected sizes of internal standard cDNA and predicted target cDNA. Expression levels of GHRe and GHRi were determined from standard curves generated. The method was sensitive enough to detect expressions down to picogram levels. Applying this method, the effect of GH and T(3) injection on GHRe and GHRi mRNA expression was determined in the liver of adult female Hisex birds and 1-day-old normal and dwarf chickens....Continue Reading

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Citations

Feb 5, 2013·General and Comparative Endocrinology·Steve Harvey
May 14, 2005·Domestic Animal Endocrinology·E DecuypereV M Darras
Mar 23, 2015·General and Comparative Endocrinology·Anna Hrabia
Jul 31, 2003·American Journal of Physiology. Endocrinology and Metabolism·O M OnagbesanE Decuypere
Jul 17, 2020·Frontiers in Physiology·Charlene HanlonGrégoy Y Bédécarrats

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