Adult female tammars of Kangaroo Island (South Australia) origin were housed in open grassed enclosures, as previously described. Care and treatment of animals conformed to the Australian National Health and Medical Research Council guidelines, and all experiments were approved by Institutional Animal Experimentation Ethics Committees. Timed pregnancies were obtained by removal of pouch young (RPY), which reactivates blastocysts in embryonic diapause. Buy Advair Diskus Online
The day of RPY is therefore designated Day 0 of gestation, with birth normally occurring 26.4 ± 1.0 days later. Pregnancy was confirmed during laparotomy, and fetal fluids (yolk sac fluid [YSF], allantoic fluid, and fetal plasma) were collected as previously described. Fluid samples were stored at -20°C until assayed. Fetal adrenal glands were dissected out and either immediately snap-frozen and stored as per fluid samples or placed directly into complete cell medium for tissue culture. Adrenal glands from an additional 14 neonates were dissected and weighed fresh on a 5-decimal-place balance.