Water soluble e2

The following chemicals and reagents were obtained from the indicated sources: Dulbecco's modified Eagle's medium (DMEM), follistatin, water-soluble E2, and penicillin-streptomycin (Sigma-Aldrich Co., St. Louis, MO); activin A and inhibin B (Abcam, Cambridge, MA); inhibin A (R&D Systems, Inc., Minneapolis, MN); and fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA).

Water-soluble E2 (encapsulated in [2-Hydroxypropyl]-β-cyclodextrin) and (2-Hydroxypropyl)-β-cyclodextrin vehicle (VEH) were purchased from Sigma-Aldrich. E2 was dissolved in saline to a final concentration of 0.02 mg/mL and administered i.p. at a dose of 0.2 mg/kg according to Gresack et al (Gresack and Frick, 2006a (link), 2006b (link)). VEH-treated mice received the same amount of β-cyclodextrin as E2-treated mice. The ERβ agonist diarylpropionitrile (DPN) and the ERα agonist 4,4’,4’’-(4-Propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT) were purchased from Tocris. Solutions were prepared in sesame oil with 10% ethanol VEH to a final concentration of 0.5 mg/ml. Volumes of 50 μL were injected s.c. for a dose of ~1 mg/kg. Cocaine hydrochloride (Mallinckrodt Pharmaceuticals) was dissolved in saline and administered i.p. at a dose of 5 mg/kg.