The proximal 350 bp of 5′-flanking sequence of the human α-subunit glycoprotein hormone gene functions in the pituitary gland, but not the placenta, in transgenic mice

To understand better the minimal DNA sequence requirements for regulated expression of the human α-subunit glycoprotein hormone gene (Hα), two lines of transgenic mice were constructed that contained a fusion gene (Hα-350CAT) consisting of only 350 bp of 5′-flanking sequence of Hα linked to the bacterial gene encoding chloramphenicol acetyltransferase (CAT). CAT activity was detectable in pituitary, but not in brain, heart, kidney, liver, lung, pancreas, or spleen in transgenic mice. Gonadectomy increased (p<0.05) CAT activity in the pituitaries of males (6.5±1.4% conversion/μg protein; mean ± SEM) and females (14.5±4.2) compared to intact males (1.2±0.3) and females (6.7±1.0). In addition, administration of a GnRH antagonist (antide; 60 μg/injection; one injection every other day) for 10 d to gonadectomized animals decreased (p<0.05) CAT activity in males (3.5±0.8) and females (2.9±0.5) compared to gonadectomized animals that received saline. Antide also reduced (p<0.05) serum concentrations of luteinizing hormone in gonadectomized males and females compared to gonadectomized animals that received saline. Surprisingly, CAT activity in the placenta of Hα-350CAT transgenic mice was not detectable (<3 SD above the mean of CAT activity in placenta from nontransgenic mice;n=77). Thus, expression of the human α-subunit promoter in the placenta of transgenic mice appears to require DNA sequences upstream of the proximal 350 bp of 5′-flanking sequence, whereas the proximal 350 bp of the human α-subunit gene contains sufficient DNA sequence to target pituitary-specific expression and confer responsiveness to gonadal hormones and GnRH.