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      Evidence for the Synthesis of Corticosteroid-Binding Globulin in Human Placenta

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          We demonstrated the expression of corticosteroid-binding globulin (CBG) in human placenta using reverse transcription-polymerase chain reaction-Southern blot analysis and immunohistochemical and immunoblotting studies. In the RT-PCR-Southern blot analysis, only one predicted PCR product was detected without nonspecific products in all samples of human placenta and 3A (tPA-30-1) human placental cells. In Western blot analysis, polyclonal anti-CBG antibodies recognized a protein of approximately 55 kD in the protein extracts prepared from 3A (tPA-30-1) cells. Additionally, CBG mRNA expression was demonstrated by in situ hybridization in the syncytiotrophoblasts. Immunohistochemical studies performed on the placenta demonstrated the presence of specific immunoreactivity in the syncytiotrophoblast layer surrounding the chorionic villi. These findings suggest that CBG is synthesized in human placenta during pregnancy in addition to its synthesis in the liver.

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          Most cited references 3

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            The plasma sex steroid binding protein (SBP or SHBG). A critical review of recent developments on the structure, molecular biology and function.

             Philip Petra (1991)
            Significant developments have taken place within the past five years on the characterization, molecular biology and function of the plasma sex steroid-binding protein, SBP (or sex hormone binding globulin, SHBG). During the span of that time, amino acid sequences of two SBPs have been established, amino acid residues in the steroid-binding site have been identified, the structure of the human SBP gene has been deduced and evidence for the possible existence of a SBP membrane receptor has been presented. This review covers the salient aspects of these and other developments including a critical analysis of the various proposed models and interpretations with regards to the structure, evolution, molecular biology and function of SBP.
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              Evidence that human placenta is a site of sex hormone-binding globulin gene expression.

              The presence of an androgen-binding component in placenta was investigated in vitro using a tissue culture system of human placental explants. Explants of trophoblastic tissue from normal term placentas were kept in culture under appropriate conditions for at least 48 h in a serum-free medium. The existence of an androgen-binding protein was explored by binding assays, immunohistochemistry studies and Northern blot analyses of placental mRNA. Steady-state polyacrylamide gel electrophoresis and Scatchard plot analyses revealed the presence of a high affinity specific binding component for 5 alpha-dihydrotestosterone in cultured placenta. Immunohistochemical studies performed on intact placenta and on Percoll-gradient purified trophoblastic cells demonstrated the presence of specific immunoreactivity in the cytoplasm of syncytial cells. Northern blot analyses of placental mRNA showed a single hybridizable 32P-labeled human sex hormone-binding globulin (SHBG) cDNA band of approx. 1.6 kb which was identical in size to that obtained with liver mRNA. The results strongly suggest the placenta as an origin of SHBG and point out this tissue as an additional site of SHBG synthesis during pregnancy.

                Author and article information

                Horm Res Paediatr
                Hormone Research in Paediatrics
                S. Karger AG
                April 1999
                27 August 1999
                : 51
                : 4
                : 162-167
                aDepartment of Obstetrics and Gynecology, and bSecond Department of Pathology, Gifu University School of Medicine, Gifu, Japan
                23351 Horm Res 1999;51:162–167
                © 1999 S. Karger AG, Basel

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                Page count
                Figures: 4, References: 38, Pages: 6
                Original Paper


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