Collateral axonal projections from rostral ventromedial medullary nitric oxide synthase containing neurons to brainstem autonomic sites

The magnocellular reticular nucleus and adjacent lateral paragigantocellular nucleus have been shown to contain a large population of nitric oxide synthase (NOS) immunoreactive neurons. However, little is known about the projections of these neurons within the central nervous system. Retrograde tract-tracing techniques combined with immunohistochemistry were used in this study to investigate whether NOS neurons in this rostral ventromedial medullary (RVMM) region send collateral axonal projections to autonomic sites in the nucleus of the solitary tract (NTS) and in the nucleus ambiguus (Amb). Fluorogold and/or rhodamine labeled latex microspheres were microinjected into the NTS and Amb at sites that elicited bardycardia and/or depressor responses (l-glutamate; 0.25 M; 10 nl). After a survival period of 10–14 days, the rats were sacrificed and tissue sections of the brainstem were processed immunohistochemically for the identification of NOS containing neuronal perikarya. After unilateral injection of the tract-tracers into the NTS and Amb, retrogradely labeled neurons were observed bilaterally throughout the RVMM region. Of the number of RVMM neurons retrogradely labeled from the NTS (684 ± 143), 9% were found to be immunoreactive to NOS. Similarly, of those RVMM neurons retrogradely labeled from the Amb (963 ± 207), 7% also contained NOS immunoreactivity. Neurons with collateral axonal projections to NTS and Amb (14% and 10%, respectively) were observed predominantly within a region of RVMM that extended co-extensively with approximately the rostrocaudal extent of the facial nucleus. Of these double labeled neurons, 36.4 ± 20 (39%) were also found to be immunoreactive to NOS. These data indicate that the RVMM contains at least three population of NOS neurons that send axons to innervate functionally similar cardiovascular responsive sites in the NTS and Amb. Although the function of these NOS containing medullary pathways in cardiovascular control is not known, it is likely that those with collateral axonal projections represent the anatomical substrate by which the RVMM may simultaneously coordinate cardiovascular responses during physiological changes associated with respiration and/or motor movements.