We examined whether corticotropin-releasing factor (CRF) was involved in orexin-induced grooming and face-washing behaviors, and whether orexin was involved in the stress reaction. Administration of alpha-helical CRF, CRF antagonist, alone had no behavioral effect, but it blocked the orexin-induced grooming and face-washing behaviors in rats. The level of corticosterone increased in a dose-dependent manner 15 min after icy injection of orexin, and it remained high for at least 60 min. In a-month-old rats, 1 h of immobilization stress increased orexin mRNA levels, but not the melanin-concentrating hormone (MCH) mRNG in the lateral hypothalamic area (LHA). In 6-month-old rats, 30 min of cold stress increased the expression of orexin mRNA in the LHA. Unlike in the 2-month-old rats, immobilization stress did not change orexin mRNA expression in g-month-old rats. These results suggest that CRF is involved in orexin-induced behaviors, and that orexin may play an important role in some stress reactions.