Physiologic and pharmacologic evidence suggests that inhibitory influences are active in the mammalian dorsal cochlear nucleus (DCN) by the onset of hearing, while anatomical evidence suggests that inhibitory synapses are not present until days or weeks later. One inhibitory neurotransmitter in the DCN is gamma aminobutyric acid (GABA) and its presence can be indexed by immunohistochemical localization of its synthetic enzyme glutamic acid decarboxylase (GAD). The present study investigated the ingrowth and synapse formation of GAD-immunoreactive inputs in the DCN of cat and hamster. GAD-immunoreactive puncta are present in the DCN of the cat at birth and of the hamster on postnatal day (PND) 3. Thus, the present data correlate well with the physiologic and pharmacologic evidence. In both species the first labelled puncta are near the dorsal acoustic stria and may originate from efferent axons in the stria. Several days later a band of labelled puncta is found in the fusiform cell layer. This location is equivalent to the termination zone of cartwheel cells, GAD-immunoreactive interneurons in the DCN. Based on this spatiotemporal sequence in the appearance of GAD-immunoreactive puncta, we suggest that sources of GABA extrinsic to the DCN mature first, followed by intrinsic sources.