We studied developmental changes in respiratory-like C4 activity and respiratory-related neurons in the ventrolateral medulla (VLM) of brainstem-spinal cord preparations from rat fetuses after embryonic day 16 (E16). In addition to respiratory nerve activity, non-respiratory activity was recorded from the C4 ventral root of preparations before E19. The burst duration of respiratory nerve discharge increased markedly at E19/20. Subtypes of neurons similar to newborn respiratory neurons were found in preparations with prolonged burst duration (more than 400 ms) after E20. These subtypes were not evident in preparations with short burst duration (less than 300 ms) before E19. About 60% of the inspiratory neurons in E17-19 preparations produced voltage-dependent burst activity, which was preserved in low Ca(2+)/high Mg(2+) synaptic blockade solution. In about 11% of the inspiratory neurons of E18-19 preparations, activation of one neuron induced activation of the inspiratory neuron network and generation of a full C4 inspiratory burst. The present findings suggest that respiratory neuron networks mature functionally to the level of the neonatal respiratory neuron networks during gestation period E19/20. Potentiation of synaptic interaction between respiratory neurons, causing developmental changes in the burst pattern, might be involved in the maturation process during late fetal stages.