Psychological stress and estrogen status are risk factors to develop painful temporomandibular joint disorders (TMJD) however the neural basis for this relationship is not known This study tested the hypothesis that repeated forced swim stress and estradiol treatment alter the phosphorylation of cAMP responsive element-binding protein (pCREB) in trigeminal subnucleus caudal's (Vc) the initial site of sensory input from the TMJ Ovariectomized female rats were given low or high dose estradiol and subjected to repeated forced swim stress for 3 days and on day 4 an intra-TMJ injection of mustard oil or vehicle was given Forced swim alone Increased the number of pCREB-positive neurons independent of estradiol treatment or TMJ stimulation in superficial and deep laminae of Vc Forced swim also increased the number of Fos-positive neurons in superficial laminae and neurokinin-1 receptor mRNA in whole dorsal Vc Independent of estradiol treatment These results indicated that persistent psychophysical stress alone was sufficient to Increase the expression of pCREB and downstream regulated genes associated with enhanced excitability in the caudal medullary dorsal horn a brainstem region thought to be critical for TMJD pain (C) 2010 Elsevier Ireland Ltd All rights reserved