Quantitative immunoelectron microscopic colocalization of GABA and enkephalin in the ventrocaudal periaqueductal gray of the rat

In the present ultrastructural study in the ventrocaudal periaqueductal gray (PAG) of the rat, the relationship and the association between GABAergic and enkephalinergic neuronal elements were investigated using postembedding colocalization immunogold electron microscopic technique in order to establish the precise relationship between these two important neurotransmitters in this part of the brain stem. The GABA-like neuronal elements were immunoreacted with 20 nm gold particles and the enkephalin (ENK)-like immunoreactive neurons were labeled with 10 nm gold particles. Double labeling of sections with ENK and GABA produced colocalization in 23.3% and 1.2% of axon terminals and dendrites, respectively. Most of the double-labeled terminals contained more GABA-like than ENK-like immunolabeling. Approximately 19.4% of the labeled axon terminals and 8.5% of the labeled dendrites contained only GABA-like immunoreactivity, while 24% of the immunolabeled dendrites were immunoreactive with only ENK-like immunoreactivity. The synapses between the two kinds of immunolabeled neuronal profiles appear to be both asymmetrical and symmetrical. GABA-like immunolabeled terminals contained small, clear, pleomorphic or round vesicles and were found to make synapses with ENK-like immunolabeled and nonimmunolabeled dendrites, whereas most of the ENK-like immunolabeled axon terminals contained dense-cored vesicles. Approximately half of the axon terminals (51%) and dendrites (56%) in the ventrolateral PAG were not labeled for either GABA or for ENK immunoreactivity. The results are discussed in terms of GABAergic inhibition of antinociceptive mechanisms in the ventrolateral PAG and of the activation of these mechanisms by ENK neurotransmitter.