Expression of growth hormone (GH)-binding protein with hydrophilic C-terminus by the mouse placenta: studies in vivo and in vitro

Barnard, R., Thordarson, G., Lopez, M.F., Yamaguchi, M., Edens, A., Cramer, S.D., Ogren, L. and Talamantes, F. (1994) Expression of growth hormone (GH)-binding protein with hydrophilic C-terminus by the mouse placenta: studies in vivo and in vitro. Journal of Endocrinology, 140 1: 125-135. doi:10.1677/joe.0.1400125


Author Barnard, R.
Thordarson, G.
Lopez, M.F.
Yamaguchi, M.
Edens, A.
Cramer, S.D.
Ogren, L.
Talamantes, F.
Title Expression of growth hormone (GH)-binding protein with hydrophilic C-terminus by the mouse placenta: studies in vivo and in vitro
Formatted title
Expression of growth hormone (GH)-binding protein with hydrophilic C-terminus by the mouse placenta: studies in vivo and in vitro
Journal name Journal of Endocrinology   Check publisher's open access policy
ISSN 1479-6805
0022-0795
Publication date 1994-01-01
Year available 1994
Sub-type Article (original research)
DOI 10.1677/joe.0.1400125
Open Access Status Not yet assessed
Volume 140
Issue 1
Start page 125
End page 135
Total pages 11
Place of publication Bristol, United Kingdom
Publisher BioScientifica
Language eng
Abstract GH-binding protein (GHBP) or GH receptor is present in numerous extrahepatic tissues in the rodent. From mid- to late gestation in the mouse, the maternal serum concentration of GHBP increases 30- to 50-fold. We have investigated whether the placenta might synthesize GHBP and potentially contribute to this increase. RNA was isolated from placentas and subjected to Northern analysis using a cDNA probe to the shared region of GHBP and GH receptor-encoding mRNAs. From day 8 to day 18 of gestation, the GHBP-encoding mRNA (1.4 kb) increased 45-fold in quantity. The GH receptor-encoding mRNA (4.2 kb) increased sixfold by day 14 and then remained steady until day 18. These changes which were not co-ordinated parallel reported changes in the steady-state concentrations of 1.4 and 4.2 kb mRNAs in maternal liver, suggesting shared regulatory factors. Extracts of freshly isolated trophoblasts were assayed for GHBP with a radioimmunoassay specific for GHBP with a hydrophilic carboxyl terminus. The cytosolic content of immunoreactive GHBP increased fourfold from mid- to late gestation. Trophoblasts were isolated from placentas and cultured for 2 days on collagen gels in defined medium. Cultured cells were at least 90% viable and secreted mouse placental lactogen-II (mPL-II). Immunocytochemistry was carried out simultaneously on cells cultured from day 7 to day 17 of gestation using a monoclonal antibody (MAb 4.3), which recognizes the hydrophilic C-terminus of GHBP. Cell-localized GHBP was present in trophoblasts cultured for 2 days, but GHBP was not detectable by radioimmunoassay or by immunoprecipitation in concentrated culture media from cultures treated with 100 ng mouse GH/ml or 100 ng mPL-II/ml or from untreated cultures. RNA was isolated from cells cultured in an identical manner to those analysed by immunocytochemistry. Three GH receptor/GHBP mRNA species of 8, 4.2 and 1.4 kb were observed. The quantity of 4.2 and 1.4 kb mRNAs did not change significantly in cultures from day 7 to day 15 of gestation but, in cultures from day 17 of gestation, the amount of 14 kb mRNA dropped significantly, while that of the 42 kb mRNA remained unchanged. GHBP- and GH receptor-encoding mRNAs are not co-ordinately regulated in vivo or in vitro. Although mPL-II was secreted into the medium by cultured trophoblasts, secretion of GHBP could not be detected. The culture medium may not contain the specific factors required for secretion of placental GHBP, or placental GHBP may not be destined for secretion.
Keyword Lactogen-Ii Secretion
Messenger-Rna
Gh-Binding
Tissue Distribution
Receptor
Serum
Rat
Purification
Pregnancy
Cloning
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID DK42361
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Chemistry and Molecular Biosciences
 
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Created: Tue, 12 Sep 2017, 20:18:57 EST by Professor Ross Barnard on behalf of School of Chemistry & Molecular Biosciences