The distribution and characterization of NADPH-d/NOS-IR neurons in the rat cuneate nucleus

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry have been used to characterize the nitric oxide (NO)-containing neurons in the rat cuneate nucleus. The present results showed that NADPH-d-positive/NOS-immunoreactive (-IR) neurons were distributed in the entire rostrocaudal extent of the nucleus. In the caudal region (approximately 1-2 mm caudal to the obex), NADPH-d/NOS-IR neurons were aggregated along the dorsal area of the nucleus notably in the lateral aspect. When traced rostrally, labeled neurons were progressively reduced and the cells were randomly distributed. The labeled neurons varied from round, ovoid to spindle-shaped with a mean profile area of about 140.1+/-1.7 microm(2) (n=720). They made up 7-10% of the neuronal population in the cuneate nucleus. By immunoelectron microscopy, the immunoreaction product was deposited throughout the cytoplasm extending from the soma to the proximal and distal dendrites. Results of NADPH-d staining paralleled that of NOS immunohistochemistry. Furthermore, NADPH-d reactivity and NOS-IR were colocalized in the same neurons following double labeling. Using NADPH-d histochemistry along with anti-gamma-aminobutyric acid (GABA) and -glycine postembedding immunolabeling for identification of GABA- and glycine-IR neurons, respectively, about 33% of the NADPH-d-positive neurons contained both GABA and glycine, 26% of them contained only glycine, while 41% of them showed neither GABA nor glycine labeling. Cuneothalamic neurons (CTNs) were identified by injecting the retrograde tracer Fluorogold (FG) into the ventrobasal complex of the thalamus. Numerous FG-labeled neurons were present in the contralateral cuneate nucleus, but none were reactive for NADPH-d. The present results suggest that approximately 60% of the NADPH-d/NOS-IR neurons in the cuneate nucleus are interneurons containing GABA and/or glycine.