The olfactory epithelium and its neuronal population are known to have a substantial capacity to recover after either direct injury or damage to the olfactory nerve. However, the mechanisms underlying that capacity for recovery, and indeed the limits on the recovery process, are not well understood. The aim of this study is to describe in detail the way in which the olfactory epithelium reconstitutes after direct injury. Adult male rats were exposed to 330 ppm methyl bromide (MeBr) gas for a single 6-hour period. The exposure destroys all of the neurons and sustentacular cells in over 95% of the olfactory epithelium of food-restricted rats and in over 90% of the epithelium in ad-libitum-fed rats of the same weight, yet substantial recovery of the olfactory epithelium occurs. In response to the lesion, cellular proliferation increases markedly beginning between 24 and 48 hours, peaks at 1 week, and persists at levels higher than the control level for more than 4 weeks after MeBr exposure. Even though proliferation accelerates promptly, the beginning of neuronal reconstitution is delayed; only a few immature neurons are observed 3 days after the lesion, yet they reappear in large numbers by the end of the first week. The first mature neurons emerge between 7 and 14 days after lesion and increase to near normal numbers by 4-6 weeks. In association with the restoration of the neuronal population, basal cell proliferation returns to control levels between 4 and 6 weeks after damage. Likewise, sustentacular cells, identifiable by anticytokeratin 18 labeling, reappear rapidly and reform a distinct lamina in the superficial aspect of the epithelium. They closely resemble their counterparts in control epithelium with regard to disposition and shape by 3 weeks after lesion and with regard to expression of olfactory-specific cytochrome P450s by 8 weeks. Thus, most areas of the epithelium are restored to a near normal appearance and cellular composition by the end of 8 weeks, suggesting that the MeBr paradigm for lesioning the epithelium offers significant advantages over techniques such as Triton X-100 or ZnSO4 irrigation. However, not all measures of epithelial status are normal even at 8 weeks. Immature neurons remain slightly more numerous than normal at this time. Furthermore, some areas of the olfactory epithelium do not recover after MeBr lesion and are replaced by respiratory epithelium.(ABSTRACT TRUNCATED AT 400 WORDS)

We have infused replication-incompetent retroviral vectors into the nasal cavity of adult rats 1 day after exposure to the olfactotoxic gas methyl bromide (MeBr) to assess the lineage relationships of cells in the regenerating olfactory epithelium. The vast majority of the retrovirus-labeled clones fall into three broad categories: clones that invariably contain globose basal cells (GBCs) and/or neurons, clones that always include cells in the ducts of Bowman's glands, and clones that are composed of sustentacular cells only. Many of the GBC-related clones contain sustentacular cells and horizontal basal cells as well. Most of the duct-related clones contain gland cells, and some also include sustentacular cells. Thus, the destruction of both neurons and non-neuronal cells that is caused by MeBr activates two distinct types of multipotent cells. The multipotent progenitor that gives rise to neurons and non-neuronal cells is a basal cell, whereas the progenitor that gives rise to duct, gland, and sustentacular cells resides within the ducts, based on the pattern of sparing after lesion and the analysis of early regeneration by using cell type-specific markers. We conclude that the balance between multipotency and selective neuropotency, which is characteristic of globose basal cells in the normal olfactory epithelium, is determined by which cell types have been depleted and need to be replenished rapidly.

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Human fetal ethanol exposure is strongly associated with ethanol avidity during adolescence. Evidence that intrauterine olfactory experience influences chemosensory-guided postnatal behaviors suggests that an altered response to ethanol odor resulting from fetal exposure may contribute to later abuse risk. Using behavioral and neurophysiological methods, the authors tested whether ethanol exposure via the dam's diet resulted in an altered responsiveness to ethanol odor in infant and adult rats. Compared with controls, (a) fetal exposure tuned the neurophysiologic response of the olfactory epithelium to ethanol odor at some expense to its responsiveness to other odorants in infantile rats-this effect was absent in adults; (b) the neural effect in infantile rats was paralleled by an altered behavioral response to ethanol odor that was specific to this odorant-this effect was also absent in adults; and (c) a significant component of the infantile behavioral effect was attributable to ethanol's effect on the olfactory neural modality. These data provide evidence for an important relationship between prenatal ethanol experience and postnatal behavioral responsiveness to the drug that is modulated or determined by olfactory function. Keywords NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptThe chemical senses are among the earliest systems to develop (Gottlieb, 1971;Schaal & Orgeur, 1992;Smotherman & Robinson, 1990), and in the chemosensory world of the uterus, they may have a unique salience. This statement is of potential significance to the field of ethanol research given that clinical and epidemiological studies provide strong data for a predictive relationship between prenatal ethanol exposure and the risk for ethanol abuse in adolescent and young adults (Alati et al., 2006;Baer, Bar, Bookstein, Sampson, & Streissguth, 1998;Streissguth, 1998;Yates, Cadoret, Troughton, Steward, & Giunta, 1998).It is well documented that olfactory function is fundamental to regulating a variety of biological processes, such as reproduction, food intake, and different social behaviors. Even communication between conspecific and heterospecific animals relies heavily on the reception and processing of odors produced by body glands and even feces and urine. Given this profound level of functional importance, not surprisingly, experience-induced plasticity in response to odorants is a means by which the olfactory system can be tuned to emphasize the transduction of stimuli that are deemed relevant within the animal's environment (Hudson, 1993(Hudson, , 1999. A variety of data indicate that the olfactory system is plastic in response to the odorant environment and have been gathered using different experimental manipulations, such as selective odorant exposure in neonates (Coopersmith & Leon, 1984;Johnson, Woo, Duong, Nguyen, & Leon, 1995;McCollum, Woo, & Leon, 1997;McLean & Harley, 2004;Sullivan & Leon, 1986;Sullivan, McGaugh, & Leon, 1991;Sullivan, Wilson, & Leon, 1989;Woo, Coopersmith, & Leon, 1987;Wo...

Human epidemiologic studies reveal that fetal ethanol exposure is highly predictive of adolescent ethanol avidity and abuse. Little is known about how fetal exposure produces these effects. It is hypothesized that fetal ethanol exposure results in stimulusinduced chemosensory plasticity. Here, we asked whether gestational ethanol exposure increases postnatal ethanol avidity in rats by altering its taste and odor. Experimental rats were exposed to ethanol in utero via the dam's diet, whereas control rats were either pair-fed an iso-caloric diet or given food ad libitum. We found that fetal ethanol exposure increased the taste-mediated acceptability of both ethanol and quinine hydrochloride (bitter), but not sucrose (sweet). Importantly, a significant proportion of the increased ethanol acceptability could be attributed directly to the attenuated aversion to ethanol's quinine-like taste quality. Fetal ethanol exposure also enhanced ethanol intake and the behavioral response to ethanol odor. Notably, the elevated intake of ethanol was also causally linked to the enhanced odor response. Our results demonstrate that fetal exposure specifically increases ethanol avidity by, in part, making it taste and smell better. More generally, they establish an epigenetic chemosensory mechanism by which maternal patterns of drug use can be transferred to offspring. Given that many licit (e.g., tobacco products) and illicit (e.g., marijuana) drugs have noteworthy chemosensory components, our findings have broad implications for the relationship between maternal patterns of drug use, child development, and postnatal vulnerability. chemosensory plasticity ͉ ethanol acceptability ͉ olfaction ͉ bitter ͉ gustation F etal ethanol exposure can have significant negative consequences for the developing human fetus. Fetal alcohol spectrum disorder describes a continuum of sequelae that can range from craniofacial malformations and mental retardation to behavioral changes such as hyperactivity and learning and memory deficits (1). There are also subtler, but nevertheless significant, clinical effects of fetal ethanol exposure. For instance, human epidemiologic studies (2, 3) report that (i) fetal ethanol exposure increases the risk of adolescent ethanol abuse, (ii) gestational exposure is the best predictor of adolescent abuse, and (iii) the earlier the age of secondary postnatal ethanol exposure, the greater the chance of long-term alcoholism. Little is known about how fetal exposure produces these changes in ethanol avidity.Numerous studies have examined how ethanol consumption during pregnancy alters the perception, preference, and consumption of ethanol by the mother's offspring. Animal studies (4, 5) have established that (i) ingested alcohol diffuses into amniotic fluid, (ii) fetuses ingest amniotic fluid and sense intrauterine ethanol, and (iii) the fetus can acquire information about ethanol-related sensory cues and display a memory of this prenatal experience. Likewise, human studies indicate that the fetus can detect and remember p...

Direct damage to the olfactory epithelium by inhalation of the olfactotoxin methyl bromide activates a population of multipotent globose basal cells, which reconstitute all depleted cell populations. Because members of the basic helix-loop-helix family of transcription factors are known to regulate neurogenesis and cell production, we performed in situ hybridization to examine the expression of several members of that family during the recovery of the rat olfactory epithelium after methyl bromide lesion. The numbers of basal cells expressing the proneural transcriptional activators Mash1, Neurogenin1, and NeuroD all fall precipitously 1 day after lesion. Mash1 levels begin to recover by 2 days, Neurogenin1 and NeuroD by 3 days, and substantial numbers of neurons reappear by 4 days. The antineurogenic factor Hes1 is limited to the sustentacular cells of the unlesioned olfactory epithelium and to the adjoining respiratory epithelium. Immediately after methyl bromide lesion, but not at any time after bulbectomy, a large fraction of residual, marker-confirmed globose basal cells initiate expression of Hes1. Subsequently, the Hes1-positive cells lose their association with the basal lamina, shift apically, and differentiate into sustentacular cells. In contrast, Hes5 is expressed by a small subset of globose basal cells and by olfactory ensheathing glia in the normal mucosa; Hes5 label disappears from both transiently after lesion. In sum, the recovery of the neuronal population after peripheral lesion recapitulates the sequence of transcription factor expression observed during embryonic development of the epithelium. Moreover, expression of Hes1 marks that population of globose basal cells committed to making sustentacular cells after methyl bromide lesion.

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.

Olfactory epithelium retains the capacity to recover anatomically after damage well into adult life and perhaps throughout its duration. None the less, olfactory dysfunctions have been reported widely for elderly humans. The present study investigates the effects of aging on the neurophysiological and anatomical status of the olfactory epithelium in barrier-raised Fischer 344X Brown Norway F1 hybrid rats at 7, 10, 25 and 32/35 months old. The posterior part of the olfactory epithelium in 32/35-month-old rats is well preserved. Globose basal cells are dividing, and new neurons are being born even at this advanced age. None the less, the numbers of proliferating basal cells and immature, GAP-43 (+) neurons are significantly decreased. Neurophysiological status was evaluated using voltage-sensitive dye techniques to assess inherent patterns of odorant-induced activity in the epithelium lining the septum and the medial surface of the turbinates. In middle and posterior zones of the epithelium, there were neither age-related changes in overall responsivity of this part of the olfactory epithelium to any of five odorants, nor shifts in the location of the odorant-induced hotspots. The inherent activity patterns elicited by the different odorants do become more distinct as a function of age, which probably reflects the decline in immature neurons and a slight, but not statistically significant, increase in mature neurons as a function of age. In contrast with the excellent preservation of posterior epithelium, the epithelium lining the anterodorsal septum and the corresponding face of the turbinates is damaged in the 32/35-month-old animals: in this part, horizontal basal cells are reactive, more basal cells and sustentacular cells are proliferating than in younger animals or in posterior epithelium of the same animals, and the neuronal population is less mature on average. Our findings indicate that degeneration of the olfactory epithelium is not an inevitable or pre-programmed consequence of the aging process, since the posterior zone of the epithelium is very well preserved in these barrier-protected animals. However, the deterioration in the anterior epithelium suggests that environmental insults can accumulate or become more severe with age and overwhelm the regenerative capacity of the epithelium. Alternatively, the regenerative capacity of the epithelium may wane somewhat with age. Either of these mechanisms or some combination of them can account for the functional and anatomical deterioration of the sense of smell associated with senescence in humans.