Inability of heterologous growth hormone (GH) to regulate GH binding protein in GH-transgenic swine.
Transgenic pigs expressing bovine, ovine, or human growth hormone (GH) structural genes fused to mouse metallothionein-I (mMT-bGH), ovine MT (oMT-oGH), or mouse transferrin (mTf-hGH) promoters were used to study the effects of GH on the regulation of serum GH-binding protein (GHBP). In the 14 transgenic pigs studied, circulating concentrations of heterologous GH ranged from 15 to 2,750 ng/mL. Using chromatographic methods, specific binding of GH was detected in serum from normal pigs but was undetectable in serum from all the transgenic pigs used, probably as a result of the high serum concentrations of heterologous GH present in these animals. Thus, to avoid interference of binding by high GH concentrations, serum samples were subjected to immunoblotting using a specific anti-GHBP antibody. A specific 54-kDa band was detected in normal pig serum as well as in sera from mMT-bGH, oMT-oGH, and mTf-hGH pigs. Additionally, sera from transgenic mMT-bGH pigs and their sibling controls were subjected to immunoprecipitation with an anti-GHBP antibody followed by immunoblotting with the same antibody. With this technique, we detected two specific bands of 53 and 45 kDa that could represent different degrees of glycosylation of GHBP. As determined by densitometric analysis the amount of GHBP in transgenic pig sera was similar to that detected in sera of the respective control animals. The amount of circulating GHBP remained unchanged even in oMT-oGH and mTf-hGH pigs that were exposed from birth to circulating concentrations of GH as high as 2,750 ng/mL. Thus, we conclude that heterologous GH do not act as modulators ofthe serum GHBP in pigs.
Cifone
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Instituto de Química y Fisicoquímica Biológicas, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
Dominici
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FP
Pursel
V G
VG
Turyn
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D000818:Animals; D030801:Animals, Genetically Modified / Q000097:blood*; D002352:Carrier Proteins / Q000097:blood / Q000187:drug effects*; D002417:Cattle; D002850:Chromatography, Gel / Q000662:veterinary; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000097:blood / Q000494:pharmacology*; D013007:Growth Hormone-Releasing Hormone / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting / Q000662:veterinary; D051379:Mice; D011863:Radioimmunoassay / Q000662:veterinary; D012756:Sheep; D013552:Swine
D002352:Carrier Proteins; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone; C051815:somatotropin-binding protein