Immuno-electron microscopic localization of the alpha(1) and beta(1)-subunits of soluble guanylyl cyclase in the guinea pig organ of corti.
Guanylyl cyclases (GC) catalyze the formation of the intracellular signal molecule cyclic GMP from GTP. For some years it has been known that the heme-containing soluble guanylyl cyclase (sGC) is stimulated by NO and NO-containing compounds. The sGC enzyme consists of two subunits (alpha(1) and beta(1)). In the present study, the alpha(1) and beta(1)-subunits were identified in the guinea pig cochlea at the electron microscopic level using a post-embedding immuno-labeling procedure. Ultrathin sections of LR White embedded specimens were incubated with various concentrations of two rabbit polyclonal antibodies to the alpha(1)- and beta(1)-subunit, respectively. The immunoreactivity was visualized by a gold-labeled secondary antibody in an energy-filtering transmission electron microscope (EFTEM). Marked immunoreactivity for both antibodies was found in the inner and outer hair cells, with numerous gold particles at the border of the cuticular plates, associated with the cell nuclei or attached to electron-dense parts of the cytoplasm. In the pillar cells and apical Deiters cells, soluble guanylyl cyclase immunoreactivity was located at the rim of the cuticular plates and between the microtubuli bundles. Together with the recently identified nitric oxide synthase isoforms [Eur. Arch. Otorhinolaryngol. 254 (1997) 396; Eur. Arch. Otorhinolaryngol. 255 (1998) 483], the soluble guanylyl cyclase may be involved in signalling processes in the organ of Corti.
Heinrich
U
U
Department of Otolaryngology - Head and Neck Surgery, Johannes Gutenberg University Medical School, 55131, Mainz, Germany.
Maurer
J
J
Koesling
D
D
Mann
W
W
Förstermann
U
U
D000818:Animals; D000906:Antibodies; D006162:Guanylate Cyclase / Q000032:analysis* / Q000276:immunology; D006168:Guinea Pigs; D006199:Hair Cells, Auditory, Inner / Q000201:enzymology* / Q000648:ultrastructure; D018072:Hair Cells, Auditory, Outer / Q000201:enzymology* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D019001:Nitric Oxide Synthase / Q000032:analysis; D015398:Signal Transduction / Q000502:physiology; D012995:Solubility; D016610:Tissue Embedding
D000906:Antibodies; D019001:Nitric Oxide Synthase; D006162:Guanylate Cyclase