We describe the location of Pre-I neurons, which are important to respiratory rhythm generation, in the rostral medulla of brainstem-spinal cord preparations isolated from newborn rats. This neuronal group was delimited in the reticular formation slightly medial to the caudal area of the facial nucleus and near the ventral surface. The effects of electrical stimulation and lesions in that region were also examined with respect to respiratory rhythm generation. Single shock stimulation induced Pre-I neuron firing and reset the phase of the respiratory rhythm. Electrolytic lesions in the Pre-I neuron region reduced the respiratory rate.