There are conflicting findings regarding the ability of estrogen to regulate TH gene expression in vivo, with some reports of elevation, reduction or no change. The basis for these discrepancies is unclear and may result from differences in dose, duration, mode of administration, or estradiol receptor (ER) subtype expressed. We found that in the nucleus of the solitary tract of OVX female rats, the response of the TH gene depends on the mode of estrogen administration. Daily injections of estradiol benzoate (EB) at 25 μg/kg for 16 days led to a greater than 2‐fold rise in TH mRNA, while administration of 17‐β estradiol (E2) (0.1 mg, 21 day release) for a similar time by pellets, triggered a 90% reduction in TH mRNA compared to vehicle treated controls. Another important parameter is the ER subtype. Many catecholaminergic cells express both ERα and ERβ. To study the mechanism and effect of ER subtypes, we co‐transfected PC12 cells with expression vectors for ERα or ERβ and TH reporter constructs. E2 elevated TH promoter driven reporter activity with ERα, while it triggered a reduction with ERβ. Further studies found that in the presence of ERα, the membrane impermeable E2‐BSA triggered a similar elevation of TH promoter activity requiring the CRE/CaRE motif. CREB and its phosphorylation at Ser133 were required for the response to E2 or E2‐BSA. The findings reveal several ways whereby estrogen can regulate TH transcription.