Expression and Regulation of Human Sex Hormone-Binding Globulin Transgenes in Mice during Development1

Abstract

Human sex hormone-binding globulin (SHBG) is produced by hepatocytes and transports sex steroids in the blood. The rat gene encoding SHBG is expressed transiently in the liver during fetal life, but it is not expressed in the liver postnatally, and the small amounts of SHBG in rat blood are derived from gonadal sources. To study the biosynthesis and function of human SHBG in an in vivo context, we have produced several lines of transgenic mice that contain either 11 kb (shbg11) or 4.3 kb (shbg4) portions of the human shbg locus. The expression and regulation of these transgenes have now been studied during fetal and postnatal development. In situ hybridization of an shbg11 transgenic mouse fetus at 17.5 days postcoitus located human shbg transcripts only in duodenal epithelial cells and hepatocytes. Temporal differences in the hepatic expression of mouse shbg and human shbg transgenes during late fetal development were reflected in corresponding differences in mouse and human SHBG levels in fetal and neonatal mouse blood. Serum concentrations of human SHBG increased during the first weeks of life regardless of gender until about 20 days of age in shbg11 mice, but after this time they continued to increase only in the males. This sexual dimorphism was reflected in corresponding differences in human SHBG messenger RNA (mRNA) abundance in the livers of these animals. However, it was not observed in shbg4 mice, in which hepatic production of plasma SHBG continued to increase after puberty regardless of gender. Serum testosterone and SHBG levels correlated in all sexually mature shbg transgenic mice. Human shbg transcripts were detectable only in testes of shbg11 mice and increased progressively in abundance from 10 days of age until the animal reached sexual maturity at 30 days of age, with appreciable increases occurring well before any changes in serum testosterone concentration. In the kidney, SHBG mRNA levels accumulated earlier in shbg11 than in shbg4 mice, and the expression of both types of transgenes was sexually dimorphic, with much higher SHBG mRNA levels in the kidneys of male mice. As increases in SHBG mRNA in the male kidneys coincided with increases in serum testosterone during sexual maturation, we reasoned that shbg transgene expression is androgen dependent in the kidney. This was confirmed by demonstrating that a decrease in SHBG mRNA abundance in male mouse kidneys after castration could be reversed by 5alpha-dihydrotestosterone treatment. Moreover, exogenous androgen increased human SHBG mRNA levels in the kidneys of female mice. In summary, comparisons of how different human shbg transgenes are expressed in vivo provides information about the positions of potential regulatory sequences that may control the hormonal regulation and tissue-specific expression of this gene during development.