We have reported that there is heterologous interaction between the mu, delta or kappa opioid receptors and the receptors for the chemokines CCL5/RANTES or CXCL12/SDF-1 in the regulation of antinociception in rats. CX3CL1/fractalkine, a chemokine that exclusively binds to CX3CR1, has been found to affect morphine analgesia and tolerance in the spinal cord. The purpose of the present study was to see if the interaction between the chemokine CX3CL1/fractalkine receptor and mu, delta or kappa opioid receptors occurs in the periaqueductal grey (PAG) of adult male S-D rats. The cold-water tail-flick (CWT) test was used to measure antinociception. The results showed that intra-PAG injection of 100 ng CX3CL1/fractalkine 30 min before administration of 400 ng DAMGO, 100 ng DPDPE or 20 microg dynorphin significantly reduced the antinociception induced by each of these peptides. These results demonstrate that activation of the CX3CL1 receptor diminishes the effect of mu, delta and kappa opioid agonists on their receptors in the PAG of rats.