Dextran backfill tracers combined with Lucifer Yellow injections for neuroanatomic studies of the leech head ganglion

摘要: Several neuronal tracing substances were applied to the cut ends of leech cephalic nerves and resulting backfills into subesophageal ganglion (sbEG) mapped. A 12 h incubation in 3 kDa dextrans conjugated either a fluorochrome or biotin (subsequently tagged with peroxidase) was satisfactory. In separate experiments, possible targets nerve afferents (R3 Retzius neurons) injected Lucifer Yellow (LY) visualize their projections. Comparison LY-R3 neuron map that dextran-backfilled D1 revealed extensive overlap sbEG. Experiments performed combining two protocols, confirming this observation. Moreover, confocal microscopy placed processes close proximity R3 processes, suggesting they could make synaptic contact one another With modifications, method be used identify such contacts using electron microscopy.