Expression of endogenous and exogenous growth hormone (GH) messenger (m) RNA in a GH-transgenic tilapia (Oreochromis niloticus)

被引:41
作者
Caelers, A
Maclean, N [1 ]
Hwang, GL
Eppler, E
Reinecke, M
机构
[1] Univ Southampton, Sch Biol Sci, Div Cell Sci, Southampton SO16 7PX, Hants, England
[2] Univ Zurich, Inst Anat, Div Neuroendocrinol, Zurich, Switzerland
关键词
absolute quantification; growth hormone; mRNA; ocean pout antifreeze promoter; transgene tilapia;
D O I
10.1007/s11248-004-5791-y
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have previously produced transgenic fish from crosses between a wild-type female tilapia (Oreochromis niloticus) and a G1 transgenic male. This line of growth-enhanced tilapia carries a single copy of a chinook salmon (s) growth hormone (GH) gene spliced to an ocean pout antifreeze promoter (OPAFPcsGH) co-ligated to a carp beta-actin/lacZ reporter gene construct, integrated into the tilapia genome. Because little is known about the expression sites of transgenes, we have characterised the gene expression patterns of sGH and tilapia (t)GH in transgenic tilapia using a newly established real-time PCR to measure the absolute mRNA amounts of both hormones. The sGH gene, which was expected to be expressed mainly in liver, was also found to be expressed in other organs, such as gills, heart, brain, skeletal muscle, kidney, spleen, intestine and testes. However, in pituitary no sGH mRNA but only tGH mRNA was found. Tilapia GH mRNA in wild-type pituitary amounted to 226 +/- 30 pg/mug total RNA but in transgenics only to 187 +/- 43 pg/mug total RNA. Liver exhibited the highest level of sGH mRNA (8.3 +/- 2.5 pg/mug total RNA) but the extrahepatic sites expressed considerable amounts of sGH mRNA ranging from 4.1 +/- 2.0 pg/mug total RNA in gills to 0.2 +/- 0.08 pg/mug total RNA in kidney. The widespread expression of the sGH gene is assumed to be due to the tissue specificity of the type III AFP gene promoter. It is assumed that our transgenic experiments, which in contrast to some other approaches caused no obvious organ abnormalities, mimick the GH expression during ontogeny. Because sGH mRNA is expressed both in liver and in extrahepatic sites it may not only promote secretion and release of liver-derived (endocrine) IGF-I leading to an overall growth enhancement but also stimulate IGF-I expression within the different organs in a paracrine/autocrine manner and, thus, further promote organ growth.
引用
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页码:95 / 104
页数:10
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