Colocalization of GABA, glycine, and their receptors at synapses in the rat spinal cord

Abstract

To determine whether GABA and glycine can act as cotransmitters at synapses in the rat spinal cord, we have compared the ultrastructural distribution of GABA(A)-receptor β3 subunit with that of the glycine receptor-associated protein gephyrin and combined this with postembedding detection of GABA and glycine. We also used a dual-immunofluorescence method to confirm that gephyrin was associated with the glycine-receptor α1 subunit throughout the cord. GABA(A) β3-subunit immunoreactivity was restricted primarily to synapses, and at a majority of these synapses the presynaptic axon was GABA-immunoreactive. Many synapses showed both GABA(A) β3 and gephyrin immunoreactivity, and at most of these synapses GABA and glycine were enriched in the presynaptic axon. These results strongly support the idea that cotransmission by GABA and glycine occurs in the spinal cord.