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Department of Biochemistry, North Carolina State University, Raleigh, North Carolina 27695-7622
Address all correspondence and requests for reprints to: Dr. William L. Miller, North Carolina State University, Department of Biochemistry, Box 7622, Raleigh, North Carolina 27695-7622. E-mail: wlmiller{at}bchserver.bch.ncsu.edu
Transgenic mice harboring the ovine FSHß (oFSHß) promoter plus first intron (from -4741 to +759 bp) linked to a luciferase reporter gene (oFSHßLuc) were generated to determine whether this promoter can direct tissue-specific expression in vivo and serve as a model for studying hormonal regulation of the FSHß gene. Of six lines of transgenic mice analyzed, luciferase was detected uniquely in the pituitaries of five of them. Pituitary luciferase activity was decreased 51–99% by chronic GnRH treatment (Lupron depot). Orchidectomy caused a 2- to 8-fold increase, and ovariectomy caused a 2- to 27-fold increase in pituitary luciferase activity. Furthermore, pituitary luciferase expression was consistently higher on estrus than on diestrus (3- to 20-fold). These data strongly suggested that the transgene was expressed specifically in pituitary gonadotropes and regulated in the same way as the endogenous mouse FSHß gene. Using primary pituitary cell cultures prepared from these transgenic mice, basal luciferase expression was maximal on day 3 and then decreased by day 6 of culture, a pattern reflected by endogenous mouse FSH secretion. In these pituitary cultures, basal oFSHßLuc expression was decreased 61–82% by follistatin or 59–79% by inhibin. Similarly, mouse FSH secretion was decreased 71% by follistatin or 65% by inhibin. Progesterone inhibited oFSHßLuc expression by 44–51%, but it had no effect on endogenous mouse FSH secretion. Estradiol lowered FSH secretion by 21%, but did not decrease oFSHßLuc expression significantly. In conclusion, these data demonstrated the ability of the oFSHß promoter to direct expression of a reporter gene specifically to pituitary gonadotropes in transgenic mice. Studying oFSHßLuc expression in vivo and in cell cultures derived from pituitaries of these transgenic mice should prove useful for understanding many features of FSHß regulation.
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