title: Alpha 2A-adrenergic receptors are present in lower brainstem catecholaminergic and serotonergic neurons innervating spinal cord.
abstract: A subtype-specific polyclonal antibody was used for the immunohistochemical detection of alpha 2A-adrenergic receptors (alpha 2A-ARs) in the rat lower brainstem (medulla and pons). Using dual-label fluorescence histochemistry, punctate alpha 2A-AR-like immunoreactivity (alpha 2A-AR-LIR) was identified in noradrenergic, adrenergic, and serotonergic neurons of the pontomedullary region. Confocal microscopic examination of material simultaneously labeled for TH-LIR and alpha 2A-LIR revealed that the clusters of alpha 2A-LIR were located intracellularly. Lower medullary neurons with spinal projections to segment T3 were retrogradely labeled using FITC-conjugated microbeads and the material was processed for simultaneous detection of alpha 2A-LIR and either TH-LIR or 5-HT-LIR. Using this triple-label approach, we found that virtually all medullary serotonergic cells (raphe pallidus, raphe obscurus and parapyramidal area) including those with identified spinal projections contain punctate alpha 2A-AR-LIR. In contrast, fewer than 10% of dorsal raphe serotonergic cells examined for comparison were immunoreactive. The triple labeling approach also indicated that more than 95% of the TH-immunoreactive cells of the dorsal and ventrolateral medulla, including those with demonstrable spinal projections (A5 noradrenergic and C1/C3 adrenergic) had detectable amounts of alpha 2A-AR-LIR. The presence of alpha 2A-ARs in a large fraction of bulbospinal pre-sympathetic neurons (noradrenergic A5, adrenergic C1 and C3 and serotonergic raphe cells) could explain the powerful and relatively selective effect of clonidine and other centrally acting alpha 2A-AR agonists on sympathetic efferent activity and hypertension.
authors.affiliation: Department of Pharmacology, University of Virginia School of Medicine, Charlottesville 22908.
authors.forename: P G
authors.initials: PG
authors.lastname: Guyenet
authors.forename: R L
authors.initials: RL
authors.lastname: Stornetta
authors.forename: T
authors.initials: T
authors.lastname: Riley
authors.forename: F R
authors.initials: FR
authors.lastname: Norton
authors.forename: D L
authors.initials: DL
authors.lastname: Rosin
authors.forename: K R
authors.initials: KR
authors.lastname: Lynch
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000033:anatomy & histology / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D008297:Male; D008526:Medulla Oblongata / Q000033:anatomy & histology / Q000166:cytology; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D011149:Pons / Q000033:anatomy & histology / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018341:Receptors, Adrenergic, alpha-2 / Q000032:analysis*; D012701:Serotonin / Q000032:analysis*; D013116:Spinal Cord / Q000166:cytology / Q000502:physiology*; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
chemical_list: D018341:Receptors, Adrenergic, alpha-2; D012701:Serotonin; D014446:Tyrosine 3-Monooxygenase