Rat Olfactory System Research Articles

Rapid down-regulation of tyrosine hydroxylase expression in the olfactory bulb of naris-occluded adult rats.

In most sensory systems, afferent innervation regulates morphological and biochemical characteristics of target cells for a limited time during development. Sensory deprivation experiments in adult rats also have suggested a critical period for afferent influences on olfactory bulb structure and function. Previous odorant deprivation studies that employed unilateral naris closure in neonatal rats demonstrated down-regulation of the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH) in dopamine neurons intrinsic to the olfactory bulb. Accompanying the altered biochemical parameters was a decrease in bulb size. To distinguish between deprivation-induced alterations in TH expression secondary to developmental sequelae and those occurring in mature neurons, the consequences of unilateral naris closure were assessed in young adult rats. In agreement with previous studies significant postnatal increases occurred in TH expression and total protein, an indication of bulb size. At 30 days post-closure, total protein was unaltered in the ipsilateral olfactory bulb but showed a small (12.9%), significant decline at 60 days. In contrast to the limited morphological consequences of odor deprivation, profound reductions occurred in TH expression. TH activity ipsilateral to the closure decreased significantly by 14 days post-closure and remained depressed for up to 6 months. In parallel with enzyme activity, TH immunoreactivity did not decline in the first few days post-closure. In situ hybridization revealed that TH mRNA levels decreased rapidly, i.e., by 2 days post-closure, reached a nadir at 1 month, and remained depressed for at least 6 months. The capacity of odor deprivation in the adult rat olfactory system to down-regulate TH expression suggests that this phenotypic alteration occurs independently of a presumed critical period.

Expression and localization of FGF-1 in the developing rat olfactory system

Primary olfactory axons project from the nasal olfactory neuroepithelium to glomeruli in the olfactory bulb where they synapse with mitral cells, the second-order olfactory neurons. We have shown that the heparin-binding growth factor FGF-1 is expressed by olfactory nerve ensheathing cells which surround fascicles of primary olfactory axons en route to the olfactory bulb. These cells are believed to modulate olfactory axon growth between the olfactory neuroepithelium and the olfactory bulb. During late embryogenesis, FGF-1 expression is turned on in the mitral cells, and the FGF-1 peptide becomes confined to layers of synaptic neuropil in the postnatal olfactory bulb. FGF-1 is selectively present in glomeruli and the external plexiform layer. In cultures of olfactory neuroepithelial cells, complexes between FGF-1 and an appropriate activating heparan sulfate proteoglycan stimulated morphological differentiation of both olfactory nerve ensheathing cells and primary sensory olfactory neurons. Thus, the spatiotemporal expression and the functional properties of FGF-1 in this system suggest that this molecule plays an important regulatory role in the formation of the olfactory pathway.

Ontogenesis of prolactin receptor gene expression in the rat olfactory system: potential roles for lactogenic hormones in olfactory development.

The PRL receptor (PRLR) is expressed at very low levels in the olfactory bulb of the adult rat but is detected in abundance in the olfactory epithelium and olfactory bulb of the fetal rat in late gestation. To explore potential roles for the lactogenic hormones in olfactory differentiation and development, we have used in situ hybridization and immunohistochemistry to examine the ontogeny of PRLR gene expression in the rat olfactory system. At midgestation (embryonic day 12.5), messenger RNAs (mRNAs) encoding the long and short isoforms of the rat PRLR were detected in the medial and lateral nasal processes, the epithelial lining of the olfactory pit, and the neuroepithelium lining the cerebral ventricles in the region of the rhinencephalon. PRLR mRNA was also expressed prominently in the frontonasal mesenchyme and in mesenchymal tissue underlying the developing brain and overlying the pontine flexure in the interpeduncular fossa. The distribution of PRLR immunoreactivity was similar to that of PRLR mRNA, indicating that the PRLR gene is translated to that of PRLR mRNA, indicating that the PRLR gene is translated to lactogenic binding protein in the rat embryo in vivo. With advancing gestation, the PRLR was expressed intensely, although discontinuously, in the olfactory epithelium and was detected in the cartilage primordia of the ethmoid, sphenoid, temporal, and mandibular bones. PRLR expression in the vomeronasal organ was confined to the luminal epithelial surface. PRLR mRNA and immunoreactive protein were first detected in the olfactory bulb on embryonic day 18. PRLR expression was most intense initially in the periventricular neuroepithelium; subsequently, robust staining of the mitral and tufted cell neurons became apparent, accompanied by intense PRLR expression in the sensory neuronal cell bodies of the olfactory epithelium. By postnatal day 5, the PRLR was expressed in abundance in mitral and tufted cells of the olfactory bulb and in neuronal cell bodies of the anterior olfactory nucleus and the piriform cortex. PRLR mRNA was also detected in the mitral cells of the olfactory bulb of the lactating rat, although at levels far lower than those in the fetal or neonatal rat. The expression of the PRLR in the olfactory system of the fetal and neonatal rat implicates novel roles for the lactogenic hormones in olfactory differentiation and development and may provide new mechanisms by which the lactogens may regulate neonatal behavior and maternal-infant interactions.

Ontogenesis of prolactin receptor gene expression in the rat olfactory system: potential roles for lactogenic hormones in olfactory development

Ontogenesis of prolactin receptor gene expression in the rat olfactory system: potential roles for lactogenic hormones in olfactory development

N-acetyl-lactosamine in the rat olfactory system: expression and potential role in neurite growth.

Primary sensory olfactory neurons exhibit a mosaic topographical projection from the olfactory neuroepithelium in the nasal cavity to the olfactory bulb formation of the telencephalon. Axons from primary neurons that are widely scattered in the epithelium terminate in discrete regions of the olfactory bulb. It has been hypothesised that carbohydrates present on the surface of primary olfactory axons mediate selective fasciculation and the formation of the topographical pathway. We examined the expression of the disaccharide N-acetyl-lactosamine in both the developing and the adult rat olfactory system. N-acetyl-lactosamine was expressed by all primary sensory olfactory neurons and by their terminations in the olfactory bulb throughout embryonic development and early postnatal life. In adults, N-acetyl-lactosamine was restricted to a subpopulation of primary sensory olfactory neurons that were dispersed throughout the neuroepithelium but that projected predominantly to the ventrolateral and ventromedial surfaces of the olfactory bulb. The axons of these neurons sort out in the outer layer of the bulb and preferentially self-fasciculate to form distinct axon bundles that terminate within select glomeruli. The role of N-acetyl-lactosamine in axon growth was tested by culturing primary sensory olfactory neurons on substrate-bound carbohydrates. Olfactory neuroepithelium cultures from both embryonic and postnatal rats revealed that substrate-bound N-acetyl-lactosamine was a strong and specific neurite growth-promoting agent. These data suggest that, during development of the olfactory projection, N-acetyl-lactosamine, which is present on all olfactory axons, acts as a nonselective permissive substrate for axon growth. In adults, however, the restricted distribution of N-acetyl-lactosamine on a subpopulation of axons may facilitate sorting out and self-fasciculation, which is necessary for preserving the mosaic nature of the olfactory pathway in this highly plastic region of the nervous system. These results support the hypothesis that cell surface carbohydrates are involved in axon growth in the olfactory system.

Effects of inhalation of cadmium on the rat olfactory system: behavior and morphology.

To investigate the effects of cadmium on olfaction, two separate studies were conducted in which male adult rats were exposed to CdO, via inhalation, for 5 h per day, 5 days a week for 20 weeks. Target exposure values of 250 and 500 micrograms/m3 were measured at 200 and 325 micrograms/m3 for the low concentration in two experiments, and 550 and 660 micrograms/m3 for the high concentration. Prior to exposure, olfactory thresholds were obtained using a conditioned suppression technique. After 20 weeks of cadmium exposure, there was no evidence of anosmia in any of the rats nor were there any significant changes observed in olfactory thresholds. Although olfaction was not impaired, cadmium levels in the olfactory bulbs of exposed rats were significantly elevated compared to controls. Cardiac and respiratory histopathology were observed at all exposure levels, but there was no evidence of nasal pathology related to exposure to cadmium. Failure of cadmium to produce olfactory dysfunction may be due to the protective effects of metallothionein and/or to the highly resilient nature of the rodent olfactory system.

Amino acid concentrations and selected enzyme activities in rat auditory, olfactory, and visual systems.

Homogenates of specific brain regions of three sensory systems (auditory, olfactory, and visual) were prepared from pigmented Long-Evans Hooded rats and assayed for amino acid concentrations and activities of glutaminase, aspartate aminotransferase (total, cytosolic, and by difference, mitochondrial), malate dehydrogenase, lactate dehydrogenase, and choline acetyltransferase. Comparing the quantitative distributions among regions revealed significant correlations between AAT and aspartate, between glutaminase and glutamate, between glutamate and glutamine, and between AAT plus glutaminase, or glutaminase alone, and the sum of aspartate, glutamate, and GABA, suggesting a metabolic pathway involving the synthesis of a glutamate pool as precursor to aspartate and GABA. Of the inhibitory transmitter amino acids, GABA concentrations routinely exceeded those of glycine, but glycine concentrations were relatively high in brainstem auditory structures.

Expression of the amyloid protein precursor of Alzheimer's disease in the developing rat olfactory system

The expression of the amyloid protein precursor (APP) of Alzheimer's disease (AD) was examined in the olfactory system of the developing rat. Two monoclonal antibodies were used to detect APP: Alz-90, which specifically recognizes APP, and 22C11 which recognizes both APP and the structurally related protein APLP-2. Very similar patterns of immunoreactivity were observed with both antibodies. APP immunoreactivity was first detected in a subpopulation of olfactory epithelial cells at embryonic day 16 (E16), at a time when primary sensory olfactory axons are first beginning to pierce the glia limitans of the olfactory bulb. At E16, there were more olfactory receptor neurons which expressed APP than the olfactory marker protein (OMP), indicating that some APP-containing neurons were not fully mature. Between E16 and postnatal day 8 (P8), there was a marked increase in the number of primary sensory olfactory neurons expressing APP. In the olfactory bulb, APP was first detected in the mitral cell layer at E18, at a time when synapses are first beginning to form between the dendrites of these cells and primary sensory axons. The level of APP detected within mitral cell perikarya decreased after birth and could no longer be detected between P3 and P8. This indicated that once synaptic connections had been initiated within olfactory glomeruli, the expression of APP within the mitral cells was down-regulated. High levels of APP were, however, detected within the olfactory nerve fiber layer and glomeruli between P3 and P8. The results demonstrate that APP expression in the olfactory system is coordinately regulated with the major periods of synaptogenesis. Since APP is a known neurite outgrowth promoting molecule, we suspect that it may play an important regulatory role in terminal sprouting during synaptogenesis in the olfactory bulb.

Identification of cells expressing galectin-1, a galactose-binding receptor, in the rat olfactory system.

Interactions between carbohydrate ligands and their receptors play an important role in cell adhesion and migration in many tissues. Cell-surface carbohydrates that contain terminal galactose have previously been implicated in primary sensory axon growth in the rodent olfactory system. The aim of the present study was to determine whether galectin-1, a galactose-binding receptor, was expressed within the rat primary olfactory pathway. Immunohistochemical and in situ hybridisation analyses revealed expression of galectin-1 by primary sensory olfactory neurons during the major embryonic period of axonogenesis as well as in maturity. In the adult olfactory bulb, galectin-1 was expressed by both second-order projection neurons and interneurons and was selectively localised to the synaptic neuropil layers. Mitral cells, the principal postsynaptic target of primary olfactory axons, began expressing this lectin soon after genesis and maintained high levels into adulthood. The expression of galectin-1 in the primary olfactory pathway and olfactory bulb neuropil suggests a role for this lectin both in the initial formation and in the subsequent maintenance of neuronal connections between the peripheral and the central olfactory neurons as well as between neurons within the bulb.

Behavioral, histological, and neurochemical effects of nickel (II) on the rat olfactory system.

Experimentally, inorganic, sulfated nickel compounds (Ni2+) have been shown to produce histological lesions in the nasal mucosa of rats, more specifically, atrophy of the olfactory epithelium. The present project was designed to assess the effects of inhalation of nickel sulfate hexahydrate on behavioral, histological, and neurochemical aspects of the olfactory system. Male Long-Evans rats were exposed to either background air (control) or 635 micrograms Ni/m3 for 16 consecutive days, 6 hr/day. Exposure resulted in selective lesions to the olfactory epithelium. The number of bipolar sensory receptor cells was slightly reduced and there was a significant decrease in the thickness of the olfactory epithelium. This was due primarily to a significant loss of the sustentacular cell population, with a thinning of the apical cytoplasm, concomitant with a reduction in the number of microvilli at the surface of these cells. Significant decreases in carnosine level, consistent with the nickel sulfate exposure, were observed. However, there were no changes in olfactory function as measured by either absolute threshold or two-oder discrimination tasks.

Expression of 9-O-acetylated gangliosides during development of the rat olfactory system.

The expression of 9-O-acetylated gangliosides recognized by the Jones monoclonal antibody (mAb) correlates with cell migration and axonal outgrowth in the developing rat nervous system. We studied the expression of these gangliosides during the development and maturation of the rat olfactory system. Beginning on embryonic day 13 (E13) the olfactory epithelium and the migratory mass were intensely stained with Jones mAb. However, though this immunoreactivity disappeared from the olfactory epithelium at E19, it remained in a few fascicles and some glomeruli of the newborn and adult olfactory bulbs. We concluded that the expression of 9-O-acetylated gangliosides by olfactory axons and/or migrating cells may facilitate axonal outgrowth during development and might be involved in the formation of new glomeruli in the mature olfactory bulb.

B-50/GAP-43 expression by the olfactory receptor cells and the neurons migrating from the olfactory placode in embryonic rats

B-50/GAP-43 is a growth-associated phosphoprotein that is commonly expressed in all developing neuronal systems. Using an immunocytochemistry approach, we have investigated the expression of this protein in the rat olfactory system during embryogenesis and neonatal development with a particular emphasis on the early developmental stages of the olfactory placode. Data show that already at embryonic day 12 (E12), a strong B-50/GAP-43 immunoreactivity was detected in few olfactory receptor cells well-recognizable by their positive short neuritic processes. The B-50/GAP-43 expression in the placodal epithelium thus appeared to coincide with the onset of neurite outgrowth. From E13 onwards, there was a rapid increase in the number of B-50/GAP-43-positive olfactory neurons and from E18, the protein was strongly expressed by nearly all neurons. In addition, results clearly demonstrate that as early as E13, B-50/GAP-43 was strongly expressed by many migrating cells which were seen leaving the pit epithelium in association with the first olfactory axons that penetrated the nasal mesenchyme. Many immunoreactive cells were also observed in the presumptive olfactory nerve layer. Experiments of double-labeling showed that B-50/GAP-43-immunostained migrating cells were also stained with anti-neuron-specific enolase (NSE). This confirms the neuronal nature of these early labeled migrating cells. The progressive disappearance of migrating neurons noted during the late stages of embryonic development is discussed in relation with their possible function in the early stages of development of the peripheral olfactory system.

Localization and regulation of low affinity nerve growth factor receptor expression in the rat olfactory system during development and regeneration

Nerve growth factor (NGF), a classic neurotrophic factor, promotes neuronal survival, maintenance, regeneration and differentiation in the peripheral nervous system and parts of the central nervous system. NGF activity is mediated by cell surface bound receptors including the low affinity NGF receptor (LNGFr) which is expressed by some peripheral and central neurons and is present on peripheral nerve Schwann cells during development and regeneration. The olfactory system is a useful model for the study of the role of LNGFr in neuronal development and regeneration. The growth of olfactory axons into the brain begins in the embryo and continues through the first few postnatal weeks. In mature animals there is persistent turnover and generation of olfactory receptor neurons (ORNs) and continuous growth of new axons into the olfactory bulb. These new axons grow along the preexisting olfactory pathway. In the mature olfactory system, LNGFr has been observed in the glomerular layer of the olfactory bulb, the target of ORNs. However, neither the cellular localization nor the developmental expression of LNGFr has been characterized. Here, we tested the hypothesis that LNGFr expression is developmentally regulated in the olfactory nerve and is reinduced following injury to the mature olfactory nerve. LNGFr-immunoreactivity (IR) was first observed in the olfactory mucosa at embryonic day (E)13 and in the olfactory nerve at E14. LNGFr-IR increased in the nerve during embryonic development, began to decrease at around postnatal day (P)5 and was scarcely detectable in normal adults. The staining pattern suggests that LNGFr is located on the olfactory nerve Schwann cells. Streaks of LNGFr-IR were present in the adult olfactory nerve. We reasoned that these streaks might represent transient reexpression of LNGFr associated with normal olfactory neuron turnover and replacement. Consistent with this hypothesis, LNGFr was robustly reexpressed in the adult olfactory nerve following lesion of the olfactory epithelium. Starting late in development (E21) and in the adult, LNGFr-IR was also observed on fibers in deep layers of the olfactory bulb. LNGFr-IR was also observed in neurons of the nucleus of the diagonal band (NDB) in the basal forebrain. NDB is the sole source of cholinergic afferents of the olfactory bulb. Thus, we tested the hypothesis that LNGFr in the deep layers of the olfactory bulb is located on NDB axons by making lesions of NDB. Following the lesion, LNGFr-IR disappeared in the deep layers of the olfactory bulb but remained in the glomerular layer.(ABSTRACT TRUNCATED AT 400 WORDS)

Rat olfactory neurons can utilize the endogenous lectin, L-14, in a novel adhesion mechanism.

L-14 is a divalent, lactosamine-binding lectin expressed in many vertebrate tissues. In the rat nervous system, L-14 expression has been observed previously in restricted neuronal subsets within the dorsal root ganglia and spinal cord. In this study we report that L-14 is expressed by nonneuronal cells in the rat olfactory nerve. We demonstrate that L-14 binds and co-localizes with two ligands in the rat olfactory system: a beta-lactosamine-containing glycolipid, and a putative member of the laminin family. The former is expressed on the surfaces of nascent olfactory axons originating from neuron cell bodies in the olfactory epithelium. The latter is present in the extracellular matrix of the axonal path leading to synaptic targets in the olfactory bulb. In vitro, we find that recombinant L-14 promotes primary olfactory neuron adhesion to two laminin family members, and promotes intercellular adhesion. Both activities are dose-dependent, and are independent of integrin-mediated mechanisms. We have thus found that L-14 can serve two distinct adhesive functions in vitro, and propose that L-14 in vivo can promote olfactory axon fasciculation by crosslinking adjacent axons and promote axonal adhesion to the extracellular matrix.

NCAM expression by subsets of taste cells is dependent upon innervation.

The expression of the neural cell adhesion molecule (NCAM) and distinct carbohydrate groups by cells of the taste buds of the rat vallate papilla was investigated by immunohistochemical and biochemical techniques. We employed antibodies against 1) the extracellular (mAb 3F4) and cytoplasmic (mAb 5B8) portions of the NCAM polypeptide, 2) the highly sialylated form of NCAM (mAb 5A5), 3) carbohydrate epitopes associated with glycosylated NCAM forms in the rat (mAb 2B8) or frog (mAb 9-OE) olfactory system, and also 4) the Lewisb blood group carbohydrate epitope (mAb CO431). NCAM mRNA was demonstrated by polymerase chain reaction (PCR) in samples of the vallate papilla, suggesting the presence of NCAM in cells of the taste buds. Antibodies against NCAM (mAbs 3F4 and 5B8) recognized a subset (about 20%) of cells within the vallate taste buds; fibers of the glossopharyngeal nerve, including those innervating the gustatory epithelium, were NCAM immunoreactive. Taste bud cells did not express polysialic acid (mAb 5A5), but mAb 5A5 immunoreactivity was observed on fibers of the IXth nerve, including a few that entered the taste buds. All or nearly all of the cells within the vallate taste buds were immunoreactive to mAb 2B8, whereas mAbs 9-OE and CO431 reacted with subsets of cells. The carbohydrates recognized by mAbs 2B8 and 9-OE were also abundantly expressed in the ducts and acini of the lingual salivary glands. Bilateral crush of the IXth nerve resulted in the loss of expression of all of these molecules from the gustatory epithelium. If cells of the taste bud express NCAM during their final stage(s) of differentiation, then NCAM could play a role(s) in the growth of gustatory axons toward their target epithelial cells and in the recognition between the nerve fibers and mature taste receptor cells, or among the taste bud cells themselves.

Synaptophysin and synaptoporin expression in the developing rat olfactory system

The expressions of two closely related synaptic vesicle antigens synaptophysin and synaptoporin were examined in the olfactory system of the adult rat and during pre- and postnatal development. In the adult, immunocytochemistry showed that the continuously regenerating olfactory receptor neurons (primary neurons) produce both synaptophysin and synaptoporin which were localized in the cell bodies of the receptor neurons in the olfactory epithelium, their dendrites, axonal processes in the olfactory nerve and their terminals in the olfactory bulb glomeruli. Furthermore, ultrastructural analysis revealed synaptophysin- and synaptoporin-immunore activities associated with synaptic vesicles in most olfactory receptor axonal terminals impinging on dendrites of the mitral and tufted neurons (secondary neurons in the olfactory bulb circuitry) in the olfactory glomeruli. In like manner, tufted neurons, granule and periglomerular neurons (interneurons in the olfactory bulb circuitry) express both synaptophysin and synaptoporin. In contrast, mitral neurons expressed only the synaptophysin antigen which was likewise associated with mitral axonal terminals in their target the olfactory cortex. The patterns of synaptophysin and synaptoporin expressions in mitral neurons (synaptophysin only) and tufted neurons (synaptophysin and synaptoporin) were similar in prenatal, postnatal and adult rats as revealed by immunocytochemistry and in situ hybridization. However, the biosynthesis of synaptophysin and synaptoporin by granule and periglomerular neurons, olfactory bulb interneurons, occurred mainly postnatally.

Olfactory bulb organ culture is supported by combined insulin-like growth factor-I and basic fibroblast growth factor

A rat olfactory bulb serum-free organ culture system has been developed, using either E19 or day 1 post-natal bulbs. Whereas E19 bulbs were self-sufficient, combined insulin-like growth factor-I (IGF-I) (100 ng/ml) and basic fibroblast growth factor (FGF) (50 ng/ml) supported growth and differentiation of day 1PN bulbs over 6 days. Morphological evidence for viability and differentiation in culture were seen, similar to that occuring in vivo, including development of glomerular layer and maintenance of mitral and tufted and granule cells. Quantitative data derived using radio-labelled amino acid or glucose incorporation supported additive or synergistic effects of IGF-I and FGF. This olfactory bulb organ culture system offers the potential for further studies of brain growth, differentiation and function in vitro.

Immunoelectron microscopic analysis of a novel carbohydrate differentiation antigen (CDA-3C2) in the developing rat olfactory and otic systems.

A carbohydrate differentiation antigen (CDA-3C2) exhibits a highly specific and restricted pattern of expression during rat embryogenesis. In the periphery of the embryo, this antigen is associated transiently with the lateral ectoderm but is retained only in the olfactory and otic epithelium throughout morphogenesis. At the light microscopic level, CDA-3C2 immunoreactivity appears mostly along cell periphery and in the extracellular matrix. The aim of the present study was to determine the specific cellular and subcellular distribution of CDA-3C2 in vivo in order to identify potential sites of cellular and tissue function of the antigen during embryogenesis. There was a strikingly similar subcellular distribution of CDA-3C2 in the developing otic and olfactory systems, found mostly along cell membranes, microvillar projections and acellular secretions of the epithelium. Mature sensory components of the epithelia were not immunoreactive, whereas supportive cells and their secreted structures were densely stained. The highly coincident nature of CDA-3C2 in both sensory epithelia suggests that this carbohydrate epitope, and possibly its carrier macromolecule, participate in a morphogenetic function common to these two sensory epithelia.

Regulation of Protein Kinase C Activity by Sensory Deprivation in the Olfactory and Visual Systems

Environmental regulation of sensory function has provided an important model of plastic mechanisms mediating neural information processing. To define potential commonalities in information processing in different systems, we investigated molecular changes elicited by sensory deprivation in the developing rat olfactory and visual systems. Protein kinase C (PKC), an intracellular messenger implicated in synaptic plasticity and memory, was analyzed. Initial, developmental studies indicated that PKC activity in the soluble and particulate fractions of the olfactory bulb increased three- to fourfold from birth to 3 months of age. Unilateral olfactory deprivation prevented the developmental increase in both soluble and particulate PKC activities in the ipsilateral olfactory bulb and piriform cortex, the second-order relay. Phorbol ester binding localized PKC to intrinsic neuronal populations and their dendrites in the control and deprived bulbs. Moreover, PKC was similarly lower in the visual cortex of dark-reared rats than in light-reared controls. The changes in PKC were region specific, as activity was unchanged by either treatment in the parietal cortex, a control area that does not process primary olfactory or visual information. Our results suggest that the important intracellular messenger, PKC, is similarly regulated in entirely different sensory systems by different environmental stimuli. Consequently, different sensory systems may use common molecular mechanisms to process information.

Olfactory afferent regulation of the dopamine phenotype in the fetal rat olfactory system

Recent studies strongly suggest that functional olfactory receptor cell innervation is necessary for the maintenance of the dopamine phenotype in the adult rat olfactory bulb. To determine whether afferent innervation is required for the initial expression of the dopaminergic phenotype during development, the current studies investigated the association between afferent innervation and phenotypic expression using both in vivo and in vitro systems. Ontogeny of the dopamine phenotype in the rat main olfactory bulb was assessed by the appearance of immunoreactivity for tyrosine hydroxylase, the first enzyme in the dopamine biosynthetic pathway. Development of receptor afferent innervation of the bulb was demonstrated with olfactory marker protein immunoreactivity. Tyrosine hydroxylase-immunoreactive cells occurred only in regions of the olfactory bulb receiving afferent innervation. However, the appearance of afferent fibers in the olfactory bulb preceded tyrosine hydroxylase expression by three to four days (gestational days 14–15 versus 18, respectively). In explant cultures, significant numbers of tyrosine hydroxylase-containing cells were observed only in en bloc co-cultures of presumptive olfactory bulb and epithelium. Explant cultures of presumptive olfactory bulb alone contained few, if any, tyrosine hydroxylase-immunoreactive cells. Similarly, expiants produced by recombining previously separated presumptive olfactory bulb and epithelium exhibited very few tyrosine hydroxylase-immunostained cells. These data suggest that expression of the dopamine phenotype, as indicated by the presence of tyrosine hydroxylase, depends on a critical level of afferent innervation. The results also support previous studies which indicated that neuronal activity or an activity-dependent process may be required for induction of tyrosine hydroxylase expression.