In rodent, there is evidence that the orienting behaviour elicited by direct stimulation of the superior colliculus (SC) is partly mediated by contralateral descending projections, while avoidance-type behaviour is associated with ipsilateral descending projections. However, the identity of target structures in the brainstem which mediate these different behavioural responses is unknown. The c-fos immediate early gene is expressed polysynaptically in neurons in response to a wide range of extracellular stimuli, and hence has been proposed as a technique for mapping functional pathways. The purpose of this study was, therefore, to use the c-fos technique to investigate the functional specificity of brainstem regions which are innervated by the two main descending projections of the SC. Patterns of fos-like immunoreactivity (FLI) were observed throughout the brainstem following electrical stimulation of the SC in Urethane-anaesthetized rats. Previously, the electrical stimulation had been shown to elicit either approach-like or avoidance-like movement. The main results of this experiment were; (i) animals in which the stimulation elicited defensive behaviour had elevated levels of immunostaining in specific terminal areas of the ipsilateral descending projections, e.g. the ventrolateral midbrain/pontine reticular formation, the cuneiform area and rostral periaqueductal grey; (ii) there was no FLI expression in any of the terminal areas of the crossed descending projection, even in animals where the electrical stimulation elicited approach. Control experiments showed that the lack of expression in the crossed descending pathway was not due to the restricted range of stimulation parameters used in the main study, or to the effects of the anaesthetic. In conclusion, this experiment was able to identify likely substrates for the mediation of defensive reactions elicited by tectal stimulation. However, given the total lack of expression in a pathway which is known to be activated, it also provides further evidence that c-fos cannot simply be used as a high resolution neuronal activity marker for mapping functional pathways.