Quantitative analysis of transient GABA expression in embryonic and early postnatal rat spinal cord neurons

GABA expression was investigated using biochemical analysis of spinal cord homogenates and immunocytochemical analysis of cells acutely dissociated from the embryonic and postnatal rat spinal cord. gamma-Aminobutyric acid (GABA) was detected by both methods as early as embryonic day 13 (E13). At E13, the percentage of neurons that were GABA+ was 0.5%. This value increased during embryogenesis, peaked during the first two postnatal weeks to just over 50%, and declined to approximately 20% by the third postnatal week emphasizing the transient nature of GABA expression. At E17 there was a pronounced, positive ventro-dorsal and rostro-caudal gradient of GABA+ cells that persisted until just before birth. At this time the gradients reversed in cervical and lumbosacral regions indicating that GABA immunoreactivity in discrete anatomical regions is also a transient phenomenon. During the embryonic period GABA immunoreactivity was diffusely distributed throughout cell bodies and proximal processes. At E21, both GABA and synaptophysin were present in the same cells. However the two antigens did not co-localize point for point. By postnatal day 21 GABA immunoreactivity appeared in puncta that co-localized entirely with puncta of synaptophysin immunoreactivity. The sizable percentage of neurons that transiently express GABA during development, and the fact that it can be detected prior to the synaptic form of glutamic acid decarboxylase (GAD65), suggest that the amino acid may play a significant role during differentiation before it functions as an inhibitory neurotransmitter.