title: Immunohistochemical assessment of cyclic guanosine monophosphate (cGMP) and soluble guanylate cyclase (sGC) within the rostral ventrolateral medulla.
abstract: Functional evidence suggests that nitric oxide (NO) signalling in the rostral ventrolateral medulla (RVLM) is cGMP-dependent and that this pathway is impaired in hypertension. We examined cGMP expression as a marker of active NO signalling in the C1 region of the RVLM, comparing adult (>18 weeks) Wistar-Kyoto (WKY, n = 4) and spontaneously hypertensive rats (SHR, n = 4). Double label immunohistochemistry for cGMP-immunoreactivity (IR) and C1 neurons [as identified by phenylethanolamine N-methyltransferase (PNMT-IR) or tyrosine hydroxylase TH-IR)], or neuronal NO synthase (nNOS) neurones, failed to reveal cGMP-IR neurons in the RVLM of either strain, despite consistent detection of cGMP-IR in the nucleus ambiguus (NA). This was unchanged in the presence of isobutylmethylxanthine (IBMX; 0.5 mM, WKY, n = 4, SHR n = 2) and in young animals (WKY, 10-weeks, n = 3). Incubation of RVLM-slices (WKY, 10-weeks, n = 9) in DETA-NO (100 mum; 10 min) or NMDA (10 muM; 2 min) did not uncover cGMP-IR. In all studies, cGMP was prominent within the vasculature. Soluble guanylate cyclase (sGC)-IR was found throughout neurones of the RVLM, but did not co-localise with PNMT, TH or nNOS-IR neurons (WKY, 10-weeks, n = 6). Results indicate that within the RVLM, cGMP is not detectable using immunohistochemistry in the basal state and cannot be elicited by phosphodiesterase inhibition, NMDA receptor stimulation or NO donor application.
authors.affiliation: Division of Health Sciences, School of Veterinary and Biomedical Science, Murdoch University, South St. Murdoch, Perth, WA, 6150, Australia.
authors.forename: Kellysan
authors.initials: K
authors.lastname: Powers-Martin
authors.forename: Anna M
authors.initials: AM
authors.lastname: Barron
authors.forename: Clare H
authors.initials: CH
authors.lastname: Auckland
authors.forename: John K
authors.initials: JK
authors.lastname: McCooke
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: McKitrick
authors.forename: Leonard F
authors.initials: LF
authors.lastname: Arnolda
authors.forename: Jacqueline K
authors.initials: JK
authors.lastname: Phillips
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism*; D006162:Guanylate Cyclase / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D008526:Medulla Oblongata / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY
chemical_list: D006162:Guanylate Cyclase; D006152:Cyclic GMP