A rich plexus of varicose fibers overlapping the C7 area of RVL, which provides the excitatory drive for tonic sympathetic discharge, may form the anatomical basis for the increases in sympathetic nerve activity provoked by systemic or central administration of cholinergic agents. (4) ChAT-immunoreactive terminals in areas where cells project to the IML support the view that central cholinergic stimulation provoking sympathoexcitation may be mediated by bulbospinal neurons. (3) Putative cholinergic terminals overlapping sites that relay primary (NTS) or higher-order visceral afferents suggest anatomical substrates for cholinergic regulation of autonomic reflexes. (2) The absence of immunoreactive elements in the LC, PVN, and SON was unexpected and suggests that cholinergic processing attributed to these nuclei is mediated polysynaptically or by synapses on processes extending into adjacent cholinoreceptor fields. Overall, these data support the concept that cholinergic agents act at multiple sites in the CNS and with topographic specificity. The role of ACh in most immunolabeled areas, however, has yet to be determined. Punctate varicosities of unknown origin and dendrites extending ventrally from the nucleus ambiguus overlapped the C1 area and immediate surround of RVL.In conclusion: 1) Cholinergic perikarya and putative terminal fields, overlap structures that are rich in cholinoreceptors and express autonomic, neuroendocrine, or behavioral responsivity to central cholinergic stimulation (PHN, NTS, RVL). In the nucleus RVL, cholinergic perikarya were in proximity to the VMS and medial to adrenergic cell bodies of the C1 area. In the NTS, previously undescribed ChAT-immunolabeled cells and processes were concentrated at intermediate and subpostermal levels and distributed viscerotopically in areas receiving primary cardiopulmonary afferents. Few or no punctate varicosities were labeled in the paraventricular (PVN) or supraoptic (SON) hypothalamic nuclei.In the mid and hindbrain, immunoreactive cells and processes were present in the nucleus of Edinger-Westphal, periaqueductal gray, parabrachial complex (PBC), a periceruleal zone avoiding the locus ceruleus (LC), pontine micturition field, pontomedullary raphe, paramedian reticular formation and periventricular gray, A5 area, lateral tegmental field, nucleus tractus solitarii (NTS), nucleus commissuralis, nucleus reticularis rostroventrolateralis (RVL), and the ventral medullary surface (VMS).In the PBC, immunoreactive varicosities identified areas previously unexplored for cholinergic autonomic responsivity superior, internal, dorsal, and central division of the lateral subnucleus, nucleus of Koelliker-Fuse and the medial subnucleus.

Cholinergic perikarya and bouton-like varicosities were localized by an immunocytochemical method empolying a monoclonal antiserum against choline acetyltransferase (ChAT), the enzyme synthesizing ACh.In the forebrain, bouton-like varicosities and/or perikarya were detected in the septum, bed nucleus of the stria terminalis, amygdala (in particular, autonomic projection areas AP1 and AP2 bordering the central subnucleus, hypothalamus rostrolateral/innominata transitional area, perifornical, dorsal, incertal, caudolateral, posterior, subparafascicular, supramammillary and mammillary nuclei.

Previously unknown neuroanatomic substrates of cholinergic - autonomic control were mapped in this study. Acetylcholine (ACh) plays a major role in central autonomic regulation, including the control of arterial blood pressure (AP).

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