Monitoring the transfection efficiency of the human folliclestimulating hormone receptor cDNA in COS-7 cells: evaluation of the growth hormone transient gene expression assay system

1996 ◽  
Vol 19 (6) ◽  
pp. 359-364 ◽  
Author(s):  
M. Simoni ◽  
J. Gromoll
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Hormone Receptor ◽  
Transfection Efficiency ◽  
Transient Gene Expression ◽  
Assay System ◽  
Gene Expression Assay ◽  
Receptor Cdna

Human growth hormone as a reporter gene in regulation studies employing transient gene expression

1986 ◽  
Vol 6 (9) ◽  
pp. 3173-3179 ◽  
Author(s):  
R F Selden ◽  
K B Howie ◽  
M E Rowe ◽  
H M Goodman ◽  
D D Moore
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Transient Expression ◽  
Human Growth Hormone ◽  
Expression System ◽  
Simian Virus 40 ◽  
Human Growth ◽  
Assay System ◽  
Transient Assay ◽  
Pituitary Cells

The human growth hormone (hGH) transient assay system described here is based on the expression of hGH directed by cells transfected with hGH fusion genes. Levels of secreted hGH in the medium, measured by a simple radioimmunoassay, are proportional to both levels of cytoplasmic hGH mRNA and the amount of transfected DNA. The system is extremely sensitive, easy to perform, and is qualitatively different from other transient expression systems in that the medium is assayed and the cells themselves are not destroyed. The hGH transient assay system is appropriate for analyses of regulation of gene expression and was utilized here to investigate the effect of the simian virus 40 enhancer on the herpes simplex virus thymidine kinase promoter and the effect of zinc on the mouse metallothionein-I promoter. The expression of hGH can also be used as an internal control to monitor transfection efficiency along with any other transient expression system. All cell types tested thus far (including AtT-20, CV-1, GC, GH4, JEG, L, and primary pituitary cells) were able to secrete hGH into the medium.

scholarly journals Dairy cows experience selective reduction of the hepatic growth hormone receptor during the periparturient period

2004 ◽  
Vol 181 (2) ◽  
pp. 281-290 ◽  
Author(s):  
J Wook Kim ◽  
RP Rhoads ◽  
SS Block ◽  
TR Overton ◽  
SJ Frank ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Growth Hormone ◽  
Dairy Cows ◽  
Hormone Receptor ◽  
Growth Hormone Receptor ◽  
Late Pregnancy ◽  
Target Tissue ◽  
Ghr Gene ◽  
Igf I

At parturition, dairy cows experience a 70% reduction in plasma IGF-I. This reduction coincides with decreased abundance of GHR1A, the liver-specific transcript of the growth hormone receptor (GHR) gene, suggesting impaired growth hormone-dependent synthesis of IGF-I. It is not immediately obvious that the periparturient reduction in GHR1A is sufficient to reduce hepatic GHR abundance. This is because approximately 50% of total GHR mRNA abundance in prepartum liver is accounted for by ubiquitously expressed transcripts which remain collectively unchanged at parturition. In addition, the possibility that parturition alters GHR expression in other growth hormone target tissue has not been examined. To address these questions, we measured GHR gene expression and GHR protein in liver and skeletal muscle of four dairy cows on days -35,+3 and+56 (relative to parturition on day 0). Hepatic GHR abundance and GHR1A transcripts were lower on day+3 than on day -35 and returned to late pregnancy value by day+56. Additional studies in two other groups of cows indicated that the hepatic levels of the GHR protein recovered substantially within 10 days after parturition. These changes occurred without variation in the abundance of HNF4, a liver-enriched transcription factor activating the promoter responsible for GHR1A synthesis. In contrast to liver, levels of GHR gene expression and GHR protein were identical on days -35,+3 and+56 in skeletal muscle. These data suggest a role for the GHR in regulating tissue-specific changes in growth hormone responsiveness in periparturient dairy cows.

scholarly journals Human growth hormone as a reporter gene in regulation studies employing transient gene expression.

1986 ◽  
Vol 6 (9) ◽  
pp. 3173-3179 ◽  
Author(s):  
R F Selden ◽  
K B Howie ◽  
M E Rowe ◽  
H M Goodman ◽  
D D Moore
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Transient Expression ◽  
Human Growth Hormone ◽  
Expression System ◽  
Simian Virus 40 ◽  
Human Growth ◽  
Assay System ◽  
Transient Assay ◽  
Pituitary Cells

The human growth hormone (hGH) transient assay system described here is based on the expression of hGH directed by cells transfected with hGH fusion genes. Levels of secreted hGH in the medium, measured by a simple radioimmunoassay, are proportional to both levels of cytoplasmic hGH mRNA and the amount of transfected DNA. The system is extremely sensitive, easy to perform, and is qualitatively different from other transient expression systems in that the medium is assayed and the cells themselves are not destroyed. The hGH transient assay system is appropriate for analyses of regulation of gene expression and was utilized here to investigate the effect of the simian virus 40 enhancer on the herpes simplex virus thymidine kinase promoter and the effect of zinc on the mouse metallothionein-I promoter. The expression of hGH can also be used as an internal control to monitor transfection efficiency along with any other transient expression system. All cell types tested thus far (including AtT-20, CV-1, GC, GH4, JEG, L, and primary pituitary cells) were able to secrete hGH into the medium.

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