Luteinising hormone (LH) and follicle stimulating hormone (FSH) comprise a common alpha-subunit and hormone-specific beta-subunit, are expressed in gonadotroph cells of the anterior pituitary and during the sheep oestrous cycle, are regulated by gonadotrophin releasing hormone (GnRH), ovarian peptides and steroids, Transcription factor steroidogenic factor-1 (SF-1) transactivates the common alpha-subunit and LH beta subunit in other species, We investigated whether SF-1 regulates beta-subunit expression and the regulation of SF-1 expression in vivo in sheep, Immunocytochemistry co-localised SF-I, LH and FSH to sheep pituitary gonadotrophs and electrophoretic mobility shift assays (EMSA) demonstrated that SF-1 bound to the LH beta promoter in vitro. NO SF-1 DNA binding site was found in the FSH beta promoter, No difference in mean levels of SF-I, FSH beta, LH beta and GnRHr mRNA was measured between the luteal and follicular phases of the oestrous cycle, However SF-1 mRNA levels were correlated to those of LH beta in individual luteal phase animals (r = 0.88, p < 0.05), when the transcription rate of LH beta was significantly higher (p < 0.01), GnRH antagonist treatment of luteal animals did not produce a significant reduction in mRNA levels of LH beta, SF-1 and FSH beta, Administering oestradiol benzoate with GnRH antagonist reduced levels of SF-1 and FSH beta mRNA from luteal values (p < 0.005), We conclude that: (1) expression of SF-1 is not solely dependent on GnRH and is downregulated by oestradiol; (2) the LH beta transcription rate is increased during the luteal phase but is not related to levels of LH beta or SF-1 mRNA; and (3) SF-1 does not appear to directly regulate expression of FSH beta, but may regulate expression indirectly, (C) 1997 Elsevier Science Ltd, All rights reserved.