title: Plasticity of adrenoceptor responsiveness on irANP secretion and pro-ANP mRNA expression in hypothalamic neuron cultures: modulation by dexamethasone.
abstract: Atrial Natriuretic Peptide (ANP) or its smaller congeners are produced and secreted from the rat hypothalamus. Whereas immunoreactive (ir)ANP secretion and proANP mRNA expression in hypothalamic cell cultures of neonatal rats were augmented by norepinephrine acting through its alpha 2-adrenoceptors (AR), in the perifusion studies of adult hypothalamic fragments beta-AR was involved in the upregulation of irANP release. Here, we report that dexamethasone (DM) modulates irANP secretion and pro-ANP mRNA expression from hypothalamic neurons in culture by switching the adrenoceptor responsiveness of the cells from alpha 2- to that of beta-AR. In long term cultures of hypothalamic cells, treatment with clonidine (alpha 2-AR agonist) increased irANP secretion in a dose related manner. This effect of clonidine was abolished by DM, a glucocorticoid which by itself had little effect on the basal release of irANP. In contrast, isoprenaline, a beta-AR agonist which was ineffective when applied alone, enhanced irANP secretion from hypothalamic cultures in the presence of DM. Concurrent incubation of DM (5 nM) and isoprenaline (10 microM) augmented irANP release approximately 3 fold above that of cultures treated with DM alone (22.6 +/- 2.2; mean +/- SE, n = 4). However, phenylephrine, an alpha 1-AR agonist alone or in the presence of DM failed to stimulate irANP release. These immunoassay findings were accompanied by corresponding changes in the abundance of pro-ANP mRNA in the cultures as examined by colorimetric Northern blot analysis employing a 30 mer oligonucleotide probe corresponding to the first 10 amino acid sequence of rANP1-28. We conclude from the above observations that glucocorticoids modulate irANP secretion and pro-ANP mRNA expression in hypothalamic neurons by altering the responsiveness of the cells from alpha 2-AR to that of beta-AR.
authors.affiliation: Neuroendocrine Laboratory, Mental Health Research Institute of Victoria, Royal Park Hospital, Parkville, Australia.
authors.forename: W
authors.initials: W
authors.lastname: Huang
authors.forename: D
authors.initials: D
authors.lastname: Lee
authors.forename: Z
authors.initials: Z
authors.lastname: Yang
authors.forename: D L
authors.initials: DL
authors.lastname: Copolov
authors.forename: A T
authors.initials: AT
authors.lastname: Lim
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D003000:Clonidine / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D009473:Neuronal Plasticity / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009638:Norepinephrine / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains; D011942:Receptors, Adrenergic, alpha / Q000187:drug effects / Q000502:physiology*
chemical_list: D011498:Protein Precursors; D012333:RNA, Messenger; D011942:Receptors, Adrenergic, alpha; D010656:Phenylephrine; D003907:Dexamethasone; D009320:Atrial Natriuretic Factor; D007545:Isoproterenol; D003000:Clonidine; D009638:Norepinephrine