Lesions of the olfactory periphery provide a means for examining the reconstitution of a diverse and highly regulated population of sensory neurons and the growth, en masse, of nascent axons to the bulb. The olfactory epithelium and its projection onto the bulb are reconstituted after ablation by methyl bromide gas, and some measure of olfactory function is restored. The extent to which the system regenerates the full repertoire of odorant receptor-expressing neurons, particularly their spatially restricted distribution across the epithelial sheet, is unknown, however, and altered odorant receptor expression might contribute to the persistent distortion of odorant quality that is observed in the lesioned-recovered animals. To address the question of receptor expression in the recovered epithelium, we performed in situ hybridization with digoxigenin-labeled riboprobes for eight odorant receptors on the olfactory epithelium from unilaterally methyl bromide-lesioned and control rats. The data demonstrate that the distribution of sensory neuron types, as identified and defined by odorant receptor expression, is restored to normal or nearly so by 3 months after lesion. Likewise, the numbers of probe-labeled neurons in the lesioned-recovered epithelium are nearly equivalent to the unlesioned side at this time. Finally, our evidence suggests that odorant receptors are distributed in multiple overlapping bands in the normal, unlesioned, and lesioned-recovered epithelium rather than in the conventionally accepted three or four zones. Thus, the primary sensory elements required for functional recovery of the olfactory system after damage are restored, and altered function implies the persistence of a more central failure in regeneration.

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The septal organ, a distinct chemosensory organ observed in the mammalian nose, is essentially a small island of olfactory neuroepithelium located bilaterally at the ventral base of the nasal septum. Virtually nothing is known about its physiological properties and function. To understand the nature of the sensory neurons in this area, we studied the mechanisms underlying olfactory signal transduction in these neurons. The majority of the sensory neurons in the septal organ express olfactory-specific G-protein and adenylyl cyclase type III, suggesting that the cAMP signaling pathway plays a critical role in the septal organ as in the main olfactory epithelium (MOE). This is further supported by patch-clamp recordings from individual dendritic knobs of the sensory neurons in the septal organ. Odorant responses can be mimicked by an adenylyl cyclase activator and a phosphodiesterase inhibitor, and these responses can be blocked by an adenylyl cyclase inhibitor. There is a small subset of cells in the septal organ expressing a cGMP-stimulated phosphodiesterase (phosphodiesterase 2), a marker for the guanylyl cyclase-D subtype sensory neurons identified in the MOE. The results indicate that the septal organ resembles the MOE in major olfactory signal transduction pathways, odorant response properties, and projection to the main olfactory bulb. Molecular and functional analysis of the septal organ, which constitutes approximately 1% of the olfactory epithelium, will provide new insights into the organization of the mammalian olfactory system and the unique function this enigmatic organ may serve.

Odorant receptors (ORs) are expressed in a spatially restricted manner in the mammalian olfactory epithelium (OE), and this patterning probably contributes to innervation specificity within the olfactory bulb (OB). Furthermore, glomerular targeting appears to be contingent on receptor choice. Central to the mechanism by which ORs influence axonal specificity is the timing of OR expression during the life cycle of the olfactory sensory neurons (OSNs). Data indicate that OSNs express ORs in the absence of the OB but do not address whether OR expression is an early event in OSN differentiation. Accordingly, we evaluated whether ORs are expressed in mature [olfactory marker protein (OMP(+))] and/or immature [growth-associated protein of 43 kDa m.w. (GAP-43(+))] OSNs by assessing the expression of the P2 OR subtype via immunostaining for beta-gal and concurrent OMP or GAP-43 expression in P2-IRES-tauLacZ mice. Nearly 90% of P2(+) OSNs expressed OMP, whereas approximately 10% expressed GAP-43. One month after unilateral bulb ablation, the number of P2(+) OSNs decreased on the lesioned side; however, the percent of P2(+)/GAP-43(+) OSNs dramatically increased. We also determined that onset of P2 OR expression is slightly delayed when evaluated in the context of neuronal differentiation. Additionally, we defined the expression of OR(+) OSNs in the OE of rats via in situ hybridization with a panel of eight ORs followed by OMP immunostaining. All eight ORs were found in neurons situated throughout the height of the OE, including those OSNs deep to OMP staining, thus demonstrating definitively that ORs are expressed prior to the maturational state defined by OMP expression.