Characterization of the corticosteroid-binding globulin messenger ribonucleic acid response in the pregnant hamster.
In this study we measured corticosteroid-binding globulin (CBG) mRNA levels in liver and various nonhepatic tissues of pregnant and nonpregnant hamsters. The N-terminal amino acid sequence (37 residues) of hamster CBG was determined and compared with published cDNA-deduced sequence information for rat and human CBG. Hamster CBG showed considerable sequence homology with both rat (70%) and human (59%) CBG. Because of the high level of homology, we were able to use a cRNA prepared from a rat CBG cDNA as a probe in Northern blot and solution hybridization analyses. Northern blots of hamster and rat liver RNA extracts revealed that the rat CBG cDNA probe hybridized to RNAs that were the same size in rats and hamsters. Further, the Northern blot showed that pregnant hamster liver contained substantially more CBG mRNA than nonpregnant hamster liver. The relative amounts of CBG mRNA in pregnant and nonpregnant hamster livers were compared using a solution hybridization assay. Slope-ratio analysis of the hybridization data revealed that pregnant hamster liver (day 14) contained 40-fold more CBG mRNA than nonpregnant hamster liver. When other tissues (kidney, spleen, small intestine, and decidual tissue) were assayed for CBG mRNA, a small amount of hybridization was detected by solution hybridization. However, Northern blot analysis of RNA extracts from nonhepatic tissues showed that the hybridizable sequences did not migrate at the same position as mature CBG mRNA. These results indicate that the observed increase in serum CBG during hamster pregnancy is largely attributable to an increase in hepatic CBG mRNA.
Department of Biological Sciences, Wichita State University, Kansas 67208.
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Lin
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Leavitt
D000595:Amino Acid Sequence; D000818:Animals; D006224:Cricetinae; D005260:Female; D006801:Humans; D008099:Liver / Q000378:metabolism; D008647:Mesocricetus; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010446:Peptide Fragments; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D015347:RNA Probes; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014156:Transcortin / Q000235:genetics*
D010446:Peptide Fragments; D015347:RNA Probes; D012333:RNA, Messenger; D014156:Transcortin