GABA nerve endings in the rat red nucleus combined detection with serotonin terminals using dual immunocytochemistry

Immunocytochemical methods were used to examine the ultrastructural features and cellular interrelationships of GABA and serotonin afferent fibers to the rat red nucleus. GABAergic nerve endings were identified in two ways, either using a pre-embedding immunoperoxidase procedure with an antibody against glutamate decarboxylase, the GABA-synthesizing enzyme, or after post-embedding immunogold labelling with an anti-GABA antibody. With the latter approach, it was possible to simultaneously visualize the GABAergic and serotoninergic innervation of the red nucleus (magnocellular part) in electron microscope preparations. This procedure involved GABA labelling of ultrathin sections obtained from specimens previously immunostained for serotonin using the pre-embedding peroxidase-antiperoxidase technique. The doubly stained sections showed gold and peroxidase markers to be present in two distinct populations of axonal varicosities. Unlike the GABAergic nerve endings, which were found to be profusely distributed throughout the nucleus, the serotonin nerve endings were relatively scarce. They contacted dendrites of large-sized neurons usually endowed with several GABA-gold labelled terminals. Not uncommonly, direct appositions between serotonin and GABA-positive terminals were also encountered. These data provide morphological evidence that red nucleus outputs may be dually regulated by GABAergic and serotoninergic afferents, while suggesting that presynaptic GABA/serotonin interactions might also play a significant part in red nucleus functions.