title: β3GnT2 maintains adenylyl cyclase-3 signaling and axon guidance molecule expression in the olfactory epithelium.
abstract: In the olfactory epithelium (OE), odorant receptor stimulation generates cAMP signals that function in both odor detection and the regulation of axon guidance molecule expression. The enzyme that synthesizes cAMP, adenylyl cyclase 3 (AC3), is coexpressed in olfactory sensory neurons (OSNs) with poly-N-acetyllactosamine (PLN) oligosaccharides determined by the glycosyltransferase β3GnT2. The loss of either enzyme results in similar defects in olfactory bulb (OB) innervation and OSN survival, suggesting that glycosylation may be important for AC3 function. We show here that AC3 is extensively modified with N-linked PLN, which is essential for AC3 activity and localization. On Western blots, AC3 from the wild-type OE migrates diffusely as a heavily glycosylated 200 kDa band that interacts with the PLN-binding lectin LEA. AC3 from the β3GnT2(-/-) OE loses these PLN modifications, migrating instead as a 140 kDa glycoprotein. Furthermore, basal and forskolin-stimulated cAMP production is reduced 80-90% in the β3GnT2(-/-) OE. Although AC3 traffics normally to null OSN cilia, it is absent from axon projections that aberrantly target the OB. The cAMP-dependent guidance receptor neuropilin-1 is also lost from β3GnT2(-/-) OSNs and axons, while semaphorin-3A ligand expression is upregulated. In addition, kirrel2, a mosaically expressed adhesion molecule that functions in axon sorting, is absent from β3GnT2(-/-) OB projections. These results demonstrate that PLN glycans are essential in OSNs for proper AC3 localization and function. We propose that the loss of cAMP-dependent guidance cues is also a critical factor in the severe axon guidance defects observed in β3GnT2(-/-) mice.
authors.affiliation: Department of Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA. timothy.henion@umassmed.edu
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Henion
authors.forename: Ashley A
authors.initials: AA
authors.lastname: Faden
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Knott
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Schwarting
mesh_terms: D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism*; D000367:Age Factors; D000606:Amino Sugars / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001369:Axons / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D019922:GAP-43 Protein / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D037102:Lectins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D017351:N-Acetylglucosaminyltransferases / Q000172:deficiency / Q000378:metabolism*; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism*; D039942:Neuropilin-1 / Q000235:genetics / Q000378:metabolism; D009831:Olfactory Mucosa / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D000099211:Phospholamban
chemical_list: D000262:Adenylyl Cyclases; D000606:Amino Sugars; D002135:Calcium-Binding Proteins; D000242:Cyclic AMP; D019922:GAP-43 Protein; D037102:Lectins; D017351:N-Acetylglucosaminyltransferases; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; D039942:Neuropilin-1; D011956:Receptors, Cell Surface; D000099211:Phospholamban; C492110:Plxna4 protein, mouse; C000458:N-acetyllactosamine; C428132:B3GNT2 protein, human; C421784:adenylate cyclase 3