Antibodies against GABA and glutamate label neurons with morphologically distinct synaptic vesicles in the locust...

Antibodies raised against GABA and glutamate were used to stain sections through locust thoracic ganglia for light and electron microscopy. Using a peroxidase-antiperoxidase method for light microscopy, the GABA antibody was shown to label inhibitory motor neurons thought to use GABA as their neurotransmitter, and the glutamate antibody to label excitatory motor neurons thought to use glutamate. An immunogold method was used to reveal labelled neuropilar processes in the electron microscope. Each antibody specifically labels a particular population of processes. With the GABA antibody, labelling is equally clear whether the processes concerned contain synaptic vesicles or not and is strongly contrasted against very low background levels. With the glutamate antibody, most processes show some affinity for the antibody, probably reflecting the presence of metabolic glutamate, however one population can be clearly distinguished by the presence of a much greater density of gold particles over synaptic vesicles. In the locust it appears, therefore, that the antibody can distinguish clearly between the metabolic and neurotransmitter pools of glutamate. It has been proposed that synaptic vesicles in GABAergic neurons have a different shape to those in glutamatergic neurons. This was supported by the electron microscope immunocytochemistry. Those showing GABA-like immunoreactivity contain predominantly pleomorphic agranular vesicles approximately 21 x 30 nm in diameter. Those showing glutamate-like immunoreactivity contain round agranular vesicles of about 38 nm in diameter. The GABA antibody appears to label all processes containing pleomorphic agranular vesicles. By contrast, some processes containing round agranular vesicles are not labelled by the glutamate antibody, even though the vesicles they contain are statistically identical in size to those in labelled profiles. With neither antibody was the labelling of glial cells greater than the background level.