The fine-structural distribution of G-protein receptor kinase 3, beta-arrestin-2, Ca2+/calmodulin-dependent protein kinase II and phosphodiesterase PDE1C2, and a Cl(-)-cotransporter in rodent olfactory epithelia.
The sequentially activated molecules of olfactory signal-onset are mostly concentrated in the long, thin distal parts of olfactory epithelial receptor cell cilia. Is this also true for molecules of olfactory signal-termination and -regulation? G-protein receptor kinase 3 (GRK3) supposedly aids in signal desensitization at the level of odor receptors, whereas beta-arrestin-2, Ca2+/calmodulin-dependent protein kinase II (CaMKII) and phosphodiesterase (PDE) PDE1C2 are thought to do so at the level of the adenylyl cyclase, ACIII. The Na+, K(+)-2Cl(-)-cotransporter NKCC1 regulates Cl(-)-channel activity. In an attempt to localize the subcellular sites olfactory signal-termination and -regulation we used four antibodies to GRK3, two to beta-arrestin-2, five to CaMKII (one to both the alpha and beta form, and two each specific to CaMKII alpha and beta), two to PDE1C2, and three to Cl(-)-cotransporters. Only antibodies to Cl(-)-cotransporters labeled cytoplasmic compartments of, especially, supporting cells but also those of receptor cells. For all other antibodies, immunoreactivity was mostly restricted to the olfactory epithelial luminal border, confirming light microscopic studies that had shown that antibodies to GRK3, beta- arrestin-2, CaMKII, and PDE1C2 labeled this region. Labeling did indeed include receptor cell cilia but occurred in microvilli of neighboring supporting cells as well. Apical parts of microvillous cells that are distinct from supporting cells, and also of ciliated respiratory cells, immunoreacted slightly with most antibodies. When peptides were available, antibody preabsorption with an excess of peptide reduced labeling intensities. Though some of the antibodies did label apices and microvilli of vomeronasal (VNO) supporting cells, none immunoreacted with VNO sensory structures.
Menco
Bert Ph M
BP
Department of Neurobiology & Physiology, O. T. Hogan Hall, Northwestern University, 2205 Tech Drive, Evanston, IL 60208-3520, USA. bertmenco@northwestern.edu
D000818:Animals; D000906:Antibodies / Q000032:analysis; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000032:analysis* / Q000276:immunology; D002923:Cilia / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D054770:G-Protein-Coupled Receptor Kinase 3; D007150:Immunohistochemistry; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008871:Microvilli / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D009831:Olfactory Mucosa / Q000737:chemistry* / Q000201:enzymology* / Q000648:ultrastructure; D010727:Phosphoric Diester Hydrolases / Q000032:analysis* / Q000276:immunology; D017346:Protein Serine-Threonine Kinases / Q000032:analysis* / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018035:Receptors, Odorant / Q000032:analysis; D028021:Sodium-Potassium-Chloride Symporters / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D018035:Receptors, Odorant; D028021:Sodium-Potassium-Chloride Symporters; D017346:Protein Serine-Threonine Kinases; D054770:G-Protein-Coupled Receptor Kinase 3; C517850:GRK3 protein, mouse; C517851:Grk3 protein, rat; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D010727:Phosphoric Diester Hydrolases