High resolution analysis of [³H]2-deoxyglucose incorporation into neurons and glial cells in invertebrate ganglia: Histological processing of nervous tissue for selective marking of glycogen

Summary

The 2-deoxyglucose (2-DG) autoradiographic technique, in conjunction with³H-labelled 2-DG and glutaraldehyde fixation, was applied to the isolated ganglia of the leech and the snail in order to analyse its potential use for the understanding of energy utilization in these simple nervous systems. Approximately 50% of the label is retained in these tissues after histological processing, and the method can be satisfactorily applied at the subcellular level. In both species most neurons progressively accumulate radioactivity over 15 min-1 h, although to different extents. In the leech the glial cells showed greater uptake than the neurons. Paired homologous neurons in contralateral buccal ganglia of the snail were equally labelled. In all structures studied by electron-microscope autoradiography the label was positively associated with glycogen. Freshly extracted glycogen from ganglia previously exposed to 2-DG was significantly labelled (2–10% of total radioactivity in the ganglia); thus the glutaraldehyde fixation did not appear responsible for artifactual binding of the label. The significance of the glycogen labelling and the potential of the technique for metabolic mapping in these nervous systems are discussed.