Expression of c-Fos in the mouse Edinger-Westphal nucleus following ethanol administration is not secondary to hypothermia or stress

Restraint stress, lipopolysaccharide (LPS), and ethanol (EtOH) administration have all been found to induce c-Fos in the brain, and to cause hypothermia. The present study was designed to assess whether the c-Fos expression that occurs in the Edinger-Westphal nucleus (EW) after EtOH administration is independent of the hypothermia or any stress effects that occur. To test this, we used restraint stress and LPS in addition to EtOH, and also examined two control areas, the dorsal raphe nucleus (DRN) and the periaqueductal gray (PAG), in addition to EW. Male C57BL6/J mice were used. Groups of mice received intraperitoneal (IP) injections of EtOH (2 g/kg), LPS (600 μg/kg or 50 μg/kg), or saline. A separate group of mice received no injection, but were placed in plastic restrainers for the entirety of the experiment. For all groups, core temperatures were monitored rectally every 30 min for 3 h postinjection, after which, the animals were sacrificed. Then, the number of Fos-positive cells in the brain regions of the EW, DRN, and PAG was quantified. Both EtOH and restraint stress induced a transient hypothermia, where core temperature (Tc) declined immediately and then rose again. Both doses of LPS induced a slower developing, longer lasting hypothermia, while saline had no effect on Tc. Only EtOH induced a significant amount of c-Fos in EW, while both doses of LPS and restraint stress induced c-Fos in DRN, and only restraint stress caused induction in PAG. These data demonstrate that activation of EW after EtOH is unrelated to hypothermia or stress.