Distribution of neuropeptidelike immunoreactivities in the guinea pig olfactory bulb

The distribution of neuropeptidelike immunoreactivities in the adult guinea pig olfactory bulb was studied immunohistochemically with antisera raised against neurotensin (NT), substance P (SP), methionine-enkephalin-Arg6-Gly7-Leu8 (ENK), somatostatin (SOM), neuropeptide Y (NPY), and cholecystokinin-8 (CCK). In the main olfactory bulb, NT-like immunoreactive (NT-IR) neurons were found among periglomerular cells. In addition, a few periglomerular cells showed ENK-like immunoreactivity. Granule cells displaying SP- or ENK-like immunoreactivities and short axon cells with SOM- or NPY-like immunoreactivities were observed in the deeper half of the granule cell layer. SOM-IR short axon cells were also seen in the external plexiform layer. Dense NT- or NPY-IR fibers were distributed in superficial lamina of the granule cell layer, and sparse SP- or CCK-IR fibers were found in the glomerular layer. In the accessory olfactory bulb, some mitral, periglomerular, and granule cells showed NT-like immunoreactivity. SP- or ENK-IR granule cells were also observed. These results are discussed in relation to laminar organization of the olfactory bulb. The most characteristic features of peptide distribution in guinea pigs, as compared with that of rats in previous studies, were the relative abundance of NT-IR structures and the lack of SP- and CCK-IR juxtaglomerular and tufted cells.     

Distribution of cholecystokinin-like immunoreactivity in the rat main olfactory bulb

The anatomical localization of cholecystokinin-like immunoreactivity (CCK-I) within the rat main olfactory bulb was analyzed by using the peroxidase-antiperoxidase immunocytochemical technique. Neurons or neuronal processes containing CCK-I were localized within all laminae of the olfactory bulb except the olfactory nerve fiber layer. A large population of CCK-I neurons, with morphology, size, and distribution corresponding to that of the middle and external tufted cells, was observed within a zone extending from the deep periglomerular region through the superficial one-half to one-third of the external plexiform layer. A smaller number of immunoreactive perikarya were found in the deep external plexiform layer, the glomerular layer, and rarely within the inner plexiform layer. These CCK-I neurons appeared to correspond to internal tufted cells, periglomerular cells, and deep short-axon cells, respectively. Dense CCK-I staining of fibers and terminals was present within the internal plexiform layer and, less densely, within the neuropil of the granule cell layer. In addition, terminal-like CCK-I was localized within layer 1A of the anterior olfactory nucleus, the olfactory tubercle, and the most rostral piriform cortex. This observation provides corroboration for the identification of the principal CCK-I neuron in the rat olfactory bulb as the centrally projecting middle tufted cell. The present results, demonstrating the localization of CCK-I to both local circuit and projection neurons of the olfactory bulb and to terminal-like puncta in the internal plexiform and granule cell layers, suggest that CCK may be significantly involved in olfactory processing at several levels.     

Distribution of D2 dopamine receptor in the olfactory glomeruli of the rat olfactory bulb

Dopamine plays key roles in the processing of the olfactory information that takes place in the olfactory glomeruli. Previous studies using autoradiography demonstrate that, at the glomerular level, these actions are mainly mediated via activation of D2 dopamine receptors. Moreover, it has been suggested that D2 receptors could be present in the olfactory nerve, where they might modulate the entrance of olfactory input into the brain. Nevertheless, the precise subcellular localization of D2 receptors in the glomerular neuropil has not been investigated. In this report, we show the subcellular distribution of D2 receptors in the glomerular circuits of Wistar rats, using pre-embedding immunogold-silver labelling and electron microscopy. Present results demonstrate for the first time the presence of D2 dopamine receptors into the terminals of the olfactory axons. In addition, we demonstrate that D2 receptors are located into presynaptic elements of the glomerular neuropil other than the olfactory axons. These elements include the dendrites of the mitral/tufted cells and the dendrites of a subset of periglomerular cells that are GABAergic and dopaminergic. This distribution pattern provides anatomical support for a wide range of actions of dopamine in the glomerular circuits through presynaptic mechanisms mediated by D2 receptors. These actions would include: (i) modulation of the glutamate release from the olfactory axons to the dendrites of mitral/tufted cells and periglomerular cells; (ii) modulation of glutamatergic synapses from the dendrites of mitral/tufted cells to the dendrites of periglomerular cells and (iii) modulation of the neurotransmission from a subset of GABAergic/dopaminergic periglomerular cells to mitral/tufted cells.     

Expression pattern of alpha CaMKII in the mouse main olfactory bulb.

Multifunctional Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is highly enriched at synapses and has been implicated in regulating the formation and function of several sensory systems, including the visual and the somatosensory systems. Although there is evidence for CaMKII expression in the olfactory system, the cellular localization of CaMKII has not been well studied and its function remains unknown. In this study, we examined the normal expression patterns of the predominant alpha CaMKII in the mouse main olfactory bulb. We showed that alpha CaMKII expression levels were high in the olfactory bulb and were developmentally regulated. Immunoreactivity to alpha CaMKII was heavy in the external plexiform layer and the granule cell layer but minimal in the olfactory nerve layer and the glomerular layer. At the cellular level, alpha CaMKII was selectively expressed in the gamma-aminobutyric acid (GABA)ergic granule cells but not in the GABAergic periglomerular cells. Unexpectedly, alpha CaMKII was not detected in the glutamatergic mitral/tufted cells. At the ultrastructural level, alpha CaMKII immunoreactivity was positive in granule cell spines and dendrites, but negative in mitral/tufted cell dendrites. In contrast, in the piriform cortex, as in the majority of cortical regions, alpha CaMKII was expressed in the glutamatergic neurons but not in the GABAergic neurons. Our results set the stage for ongoing investigations of the roles of CaMKII in the formation and function of the olfactory system.     

Cell type-dependent expression of HCN1 in the main olfactory bulb

In many brain regions, hyperpolarization-activated cationic currents (Ih) are involved in the generation of rhythmic activities, but the role of Ih in olfactory oscillations remains unclear. Knowledge of the cellular and subcellular distributions of hyperpolarization-activated and cyclic nucleotide-gated channel (HCN) subunits is necessary for understanding the role of Ih in olfactory network activities. Using light microscopic immunocytochemistry, we demonstrate strong HCN1 labelling of the glomerular layer and moderate staining of granule cell, internal and external plexiform layers of the rat main olfactory bulb. In the glomerular layer, among many unlabelled neurons, two distinct subpopulations of juxtaglomerular cells are labelled. Approximately 10% of the juxtaglomerular cells strongly express HCN1. These small diameter cells are immunoreactive for GABA and comprise a subpopulation of periglomerular cells. An additional subset of juxtaglomerular cells ( approximately 1%) expresses low levels of HCN1. They are large in diameter, GABA immunonegative but immunopositive for vesicular glutamate transporter 2, characterizing them as external tufted cells. Quantitative immunogold localization revealed that the somatic plasma membranes of periglomerular cells contain approximately four times more HCN1 labelling than those of external tufted cells. Unlike in cortical pyramidal cells, immunogold density for HCN1 does not significantly differ in somatic and dendritic plasma membranes of external tufted cells, indicating that post-synaptic potentials arriving at proximal and distal dendrites are modulated by the same density of Ih. Our results demonstrate a cell type-dependent expression of HCN1 in the olfactory bulb and predict a differential contribution of distinct juxtaglomerular cell types to network oscillations.     

Subcellular localization of m2 muscarinic receptors in GABAergic interneurons of the olfactory bulb

We analysed the ultrastructural distribution of the m2 muscarinic receptor (m2R) in the rat olfactory bulb (OB) using immunohistochemical techniques and light and electron microscopy. m2R was differentially distributed within the cellular compartments of gamma-aminobutyric acid (GABA)ergic bulbar interneurons. It is located in the gemmules of granule cells and in the synaptic loci of the interneurons of the external plexiform layer, suggesting that m2R activation could modulate the release of GABA from these interneurons onto principal cells by a presynaptic mechanism. By contrast, the receptor appears in the somata and dendritic trunks of second-order short-axon interneurons located in the inframitral layers, suggesting that postsynaptic muscarinic activation in these cells could elicit the inhibition of granule cells, leading to a disinhibition of principal cells. We also detail the anatomical substrate for a new putative muscarinic modulation that has not been previously described, and that could influence the reception of sensory information within the olfactory glomeruli. m2R appears in a subset of GABAergic/dopaminergic juxtaglomerular cells innervated by olfactory axons but is absent in juxtaglomerular cells that do not receive sensory inputs. This finding suggests that m2R activation could modify, through dopaminergic local circuits, the strength of olfactory nerve inputs onto principal cells. Activation of the muscarinic receptor may modulate the olfactory information encoding within olfactory glomeruli and may facilitate the bulbar transmission to superior centres influencing the GABA release by presynaptic and postsynaptic mechanisms. Taken together, our data provide the neuroanatomical basis for a complex action of m2R at different levels in the mammalian OB.     

Development of olfactory marker protein and tyrosine hydroxylase immunoreactivity in the transplanted rat olfactory bulb.

Evidence suggests that tyrosine hydroxylase (TH) expression by juxtaglomerular (JG) neurons of the olfactory bulb (OB) is dependent upon input from primary olfactory neurons (ONs), which are identifiable using immunocytochemical localization (ICC-L) methods for olfactory marker protein (OMP). When the input from the continuously regenerating ONs is temporarily removed (either surgically or chemically), JG cells cease TH production until ON contact is reestablished. We are studying this transneuronal regulation using the rat OB in a transplantation (TX) model. Fetal OBs, labeled in utero with tritiated thymidine, were transplanted (TX) into a site vacated by removal of a neonatal host OB. Host animals were sacrificed at varying periods after TX. Alternate sets of frozen sections were then processed for autoradiography or using ICC-L for TH and OMP. As early as 1 week post-TX, OMP-positive fibers and glomerulus-like structures were seen throughout the TX OB. Despite this extensive and rapid OMP reinnervation, TH expression returned very slowly and the number of TH expressing cells never approached control levels. The reduced TH activity in TXs may be due to failure of JG cells to survive or to develop the correct phenotype under TX conditions. Alternatively, input from ON fibers may only be necessary, but not sufficient, for the expression of TH.     

Substance P and catecholaminergic expression in neurons of the hamster main olfactory bulb

A coordinated series of immunohistochemical and biochemical analyses have been conducted in the hamster to examine the dependence of substance P and tyrosine hydroxylase (TH) expression by second-order olfactory neurons, and the level of dopamine in the main olfactory bulb (MOB), on the integrity of carnosine- and olfactory marker protein (OMP)-containing primary afferent neurons. Substance P-like immunoreactivity (SPLI) is localized in external tufted cells and centrifugal afferents of the MOB; TH immunoreactivity has a wider distribution, in external tufted, middle tufted, periglomerular, and deep short-axon cells as well as in centrifugal afferents. To characterize the SPLI, this material was isolated by guanidine-HCl extraction and passage over a C18 SEP-PAK. The SPLI coelutes on HPLC with authentic substance P and, following oxidation, coelutes with substance P sulfoxide. It is sensitive to alpha-chymotrypsin and is resistant to trypsin. Thus, the SPLI in the MOB behaves as authentic substance P. Intranasal irrigation with 0.17 M ZnSO4 results in peripheral deafferentiation of the MOB for up to 8 months as evidenced by a persistent loss of OMP immunoreactivity and shrinkage of the olfactory nerve layer and glomeruli. By these criteria, the vomeronasal inputs to the accessory olfactory bulb are not destroyed and the spared vomeronasal receptor neurons do not innervate the vacated peripheral projection field in the MOB. The loss of peripheral inputs to the MOB is accompanied by marked and parallel reductions in the incidences of SPLI- and TH-positive second-order neurons despite an increase in the density of neuronal somata in the glomerular layer. Biochemical quantifications following peripheral deafferentation also demonstrate significant decreases of both substance P and dopamine, together with the expected decrease of carnosine. In contrast, the SPLI and the TH and serotoninlike immunoreactivities in centrifugal afferents as well as the TH immunoreactivity in deep interneurons do not appear to be reduced, and the MOB content of norepinephrine in centrifugal afferents is unaffected. These results collectively indicate that the loss of inputs from the primary olfactory receptor neurons can reduce the levels of at least two different, putatively neuroactive compounds (substance P and dopamine) in at least three classes of second-order neurons (external tufted, middle tufted, and periglomerular cells). The control of central neuron phenotype by the peripheral olfactory neurons thus appears to be a phenomenon of broad influence. It may play a role in processing chemosensory information as well as offering a system in which to study neuronal plasticit     

A quantitative study of the effects of early unilateral olfactory deprivation on the number and distribution of mitral and tufted cells and of glomeruli in the rat olfactory bulb

Anatomic effects of early unilateral olfactory deprivation on the developing olfactory bulb were investigated in the albino rat. Unilateral anosmia was experimentally induced by neonatal cauterization of the left or right nare; regenerating tissue permanently blocked the nostril. The anosmic olfactory bulb (ipsilateral to the blocked nare) and its contralateral counterpart, serving as the normal control, were compared for the following quantitative anatomic parameters: total number and distribution of mitral and tufted cells and olfactory glomeruli; average diameters of mitral cells and glomeruli; relative dimensions of the bulb and its layers. The effects on mitral cells and glomeruli were studied at the ages of 25 and 60 days and at 2 years; other studies were carried out in the 25-day-old animals only. In the normal mature bulb, the number of mitral cells, tufted cells and glomeruli was found to be about 70,000, 160,000 and 3000, respectively. It was found that the absence of early olfactory stimulation was invariably correlated with a significant and permanent loss of mitral cells, amounting to more than a quarter of the total number. This loss apparently occurred rapidly during the first 3 week, as it was already evident by day 25 and did not increase appreciably with prolongation of deprivation. Tufted cells were apparently even more susceptible, because their number decreased by about 45%. As evident from their distribution profiles, the loss of mitral and tufted cells occurred uniformly throughout the bulb. It is shown that these differences were due neither to inherent interbulbar differences, nor to a hyperplasia in the normal bulb. Early anosmia had no significant effects on the number or average diameter of the glomeruli. Morphometric studies revealed that the dimensions and thickness of layers (internal and external plexiform and granular) of the anosmic bulb were significantly reduced. It is suggested that early olfactory stimulation is necessary for survival of the developing mitral and tufted cells; thus the first 2–3 postnatal weeks, covering the final developmental stages of these cells, would constitute a vulnerable period in the development of the rat olfactory system.     

Golgi analyses of dendritic organization among denervated olfactory bulb granule cells

In the vertebrate olfactory bulb, the primary projection neurons, mitral and tufted cells, have reciprocal dendrodendritic synapses with respective subpopulations of anaxonic interneurons called granule cells. In the neurological murine mutant Purkinje Cell Degeneration (PCD), all mitral cells are lost during early adulthood. As a consequence, a subpopulation of granule cells is deprived of both afferent input and efferent targets. The effect of this event on the morphology and sublaminar distribution of granule cells was studied with light microscopic Golgi procedures in affected homozygous recessive PCD mutants and normal heterozygous littermate controls. In the control mice, a minimum of three subpopulations were identified predominantly on the basis of the topology of apical dendrites and their spinous processes within the external plexiform layer (EPL) of the olfactory bulb: type I had dendrites extending across the full width of the EPL and a homogeneous distribution of spines; type II had dendritic arbors confined to the deeper EPL; type III had apical dendrites that arborized extensively within the superficial EPL with no arbors or spines present in the deeper EPL. Prior studies suggest that type II cells form connections with mitral cells; type III cells form connections with tufted cells; and type I cells may integrate information from both populations of projection neurons. In the mutant PCD mice, the classification of subpopulations of granule cells proved difficult due to a compression of dendritic arbors within the EPL. Dendritic processes followed a more horizontal tangent relative to the radial orientation seen in control mice. The length of dendritic branches was reduced by approximately 20% with a corresponding decrease in the number of spines. The density of spines (#/1 micron of dendrite) was constant in both controls and mutants at approximately 0.21. Truncation of the dendrites in the PCD mutants appeared to occur at terminal portions because the number of dendritic bifurcations was equal in both groups of mice. The data are discussed in terms of subpopulations of granule cells in the mouse olfactory bulb, the sublaminar organization of olfactory bulb circuits, and the capacity for survival and plasticity in the reciprocal dendrodendritic circuits mediated by the granule cell spines.     

Expression of PTPRO in the interneurons of adult mouse olfactory bulb

PTPRO is a receptor‐type protein tyrosine phosphatase (PTP) with a single catalytic domain in its cytoplasmic region and multiple fibronectin type III‐like domains in its extracellular region. In the chick, PTPRO mRNA has been shown to be particularly abundant in embryonic brain, and PTPRO is implicated in axon growth and guidance during embryonic development. However, the temporal and spatial expression of PTPRO protein in the mammalian CNS, particularly in the juvenile and adult mammalian brain, has not been evaluated in any detail. By immunohistofluorescence analysis with a monoclonal antibody to PTPRO, we show that PTPRO is widely expressed throughout the mouse brain from embryonic day 16 to postnatal day 1, while expression is largely confined to the olfactory bulb (OB) and olfactory tubercle in the adult brain. In the OB, PTPRO protein is expressed predominantly in the external plexiform layer, the granule cell layer, and the glomerular layer (GL). In these regions, expression of PTPRO is predominant in interneurons such as γ‐aminobutyric acid (GABA)‐ergic or calretinin (CR)‐positive granule cells. In addition, PTPRO is expressed in GABAergic, CR‐positive, tyrosine hydroxylase‐positive, or neurocalcin‐positive periglomerular cells in the GL. Costaining of PTPRO with other neuronal markers suggests that PTPRO is likely to be localized to the dendrites or dendritic spines of these olfactory interneurons. Thus, PTPRO might participate in regulation of dendritic morphology or synapse formation of interneurons in the adult mouse OB. J. Comp. Neurol. 518:119–136, 2010. © 2009 Wiley‐Liss, Inc.     

Evidence for coexistence of GABA and dopamine in neurons of the rat olfactory bulb

Immunoreactivities for gamma-aminobutyric acid (GABA) and the dopamine-synthesizing enzyme tyrosine hydroxylase (TH) were localized ultrastructurally and colocalized at the light microscopic level in neurons of the rat main olfactory bulb. By means of a simultaneous indirect immunofluorescence technique, GABA and TH immunoreactivities were found to coexist in a large number of neurons in the glomerular and external plexiform layers. Virtually all the TH-immunoreactive periglomerular neurons also contained GABA immunoreactivity (GABA-I) while there was an additional number of GABA-immunoreactive periglomerular cells (27%) which did not contain TH immunoreactivity (TH-I). In contrast, the numerous tufted-type neurons in the glomerular and superficial external plexiform layers which contained TH-I did not contain GABA-I. In the external plexiform layer (EPL), 41% of the immunoreactive neurons contained GABA-I alone, 24% contained TH-I alone, and 35% contained both. EPL neurons containing GABA-I only or both GABA-I and TH-I never exhibited tufted cell morphological characteristics and were generally of the short-axon type. Electron microscopic examination of GABA-I and TH-I elements in the glomerular layer detected morphologically similar periglomerular perikarya and intraglomerular processes immunoreactive for each substance and other neurons and processes of the same type containing neither GABA-I or TH-I. These data indicate that the classical neurotransmitters GABA and dopamine coexist in large numbers of neurons in the rat main olfactory bulb including characteristic periglomerular cells and certain other local-circuit neuronal types.     

Organization of glomeruli in the main olfactory bulb of Xenopus laevis tadpoles

Structural and functional investigations were carried out to study olfactory glomeruli in the main olfactory bulb (OB) in tadpoles of the clawed frog, Xenopus laevis. Calcium imaging of odor response patterns of OB neurons revealed that the synapses within the glomeruli are functional. Tracing axons of individual olfactory receptor neurons (ORNs), dendrites of mitral/tufted (M/T) cells and processes of periglomerular interneurons indicate that the glomerular architecture is solely determined by terminal branches of ORN axons and tufts of M/T primary dendrites. The small population of periglomerular neurons forms wide-field arborizations that always extend over many glomeruli, enter the glomeruli, but lack any glomerular tufts. Antibodies to synaptophysin indicate a high density of synapses within glomeruli, which was further confirmed at the ultrastructural level and quantified to approximately 0.5 synaptic sites per microm(2). Combining immunocytochemistry and ultrastructural investigations, we show that glomeruli in Xenopus laevis tadpoles lack any cellular borders. Glomeruli are surrounded neither by periglomerular somata nor by glial processes. Taken together, our results demonstrate that olfactory glomeruli in Xenopus laevis tadpoles (1) are fully functional, (2) are spheroidal neuropil aggregations of terminal tufts of ORNs and tufts of primary dendrites of M/T cells, and (3) are not enwrapped by a border formed by juxtaglomerular cells.     

Morphological variations among output neurons of the olfactory bulb in the frog (Rana ridibunda).

Morphological properties of putative output cells have been studied in detail in the olfactory bulb of frogs (Rana ridibunda). Intracellular injection of Lucifer Yellow was used to reconstruct individual neurons. Ten different anatomical features related to cell shape and position were studied quantitatively. The results show that output cells, generally considered to be a homogeneous group in the olfactory bulb of amphibians, are, in fact, quite different in their morphology. Using multidimensional analysis to examine differences among the output neurons, we found that they might be divided into at least two groups. In one group, the cell somata were located near the glomerular layer and the dendrites lay at large angles with respect to each other. In the other group, the cell somata were farther from the glomerular layer and their dendrites lay at smaller angles. From their morphology, these two cell groups appear to be homologous, respectively, to the superficial/middle tufted cells and deep tufted/mitral cells of mammals.     

Species-specific distribution of aromatic L-amino acid decarboxylase in the rodent adrenal gland, cerebellum, and olfactory bulb

Aromatic L-amino acid decarboxylase (AADC), the enzyme that converts L-dopa to dopamine, displayed species-specific differences in both activity and immunoreactivity in the cerebellum, olfactory bulb, and adrenal glands of three rodent species, the hamster, rat, and mouse. Specifically, in the hamster but not the rat or mouse, AADC immunoreactive cells were observed in the cerebellum and adrenal cortex. The unusual distribution of the enzyme was confirmed biochemically. AADC activity was greater in the adrenal gland and the cerebellum in the hamster than in the mouse or rat. In addition, by Western blot analysis, one band of appropriate molecular weight was observed both in the hamster adrenal gland and cerebellum. The rat adrenal gland displayed a similar immunoreactive protein on the Western blot; however, the protein could not be detected in the rat cerebellum by the technique utilized. Tyrosine hydroxylase (TH) immunoreactivity in these same tissues did not differ among the species. In the main olfactory bulb of the mouse, juxtaglomerular cells exhibited very limited immunoreactivity for AADC, but TH-immunoreactivity in these cells was robust. In contrast, juxtaglomerular cells in the rat displayed a similar intensity of immunostaining for both AADC and TH. AADC activity in the mouse, consistent with the reduced immunostaining for the enzyme, was 50% of that in the rat and the hamster. These data demonstrate that AADC protein, which is contained in cells of diverse function, also displays qualitative and quantitative species specific variations in both distribution and amount.     

Localization of 5-HT2A receptor mRNA by in situ hybridization in the olfactory bulb of the postnatal rat

The olfactory bulb receives a dense serotonergic input and appears to require serotonergic input in early olfactory associational learning. However, it is not known which cell types receive the serotonergic input or whether the cells express markers for the input throughout life. These issues need resolution in order for the mechanisms of serotonergic interactions to be better understood. The mRNA for the 5-HT_2A receptor was localized in the olfactory bulb of postnatal day 1, 2, 14 and 9-month-old Sprague-Dawley rats as well as in the bulb of adult (6 months) and aged (22-30 month) Fisher 344 rats by in situ hybridization using an ³⁵S-labelled 5-HT_2A specific oligonucleotide probe mixture. In all animals, hybridization was observed in mitral cells which are the major output cells of the main olfactory bulb. Tufted cells, located in the external plexiform layer and juxtaglomerular region, were readily observed in adult and aged animals and were also observed, albeit not as readily, in neonate pups. Quantitative analysis of the silver grain density over cells confirmed qualitative observations and showed that mitral and tufted cells were labelled in the neonate as well as in adult and aged animals. Labelled cells were also numerous in the external division of the anterior olfactory nucleus in all animals. 5-HT_2A receptor mRNA could not be detected either qualitatively or quantitatively by in situ hybridization in the accessory olfactory bulb, nor could it be observed in the olfactory epithelium. The results suggest that the output cells of the main olfactory bulb receive serotonergic input via 5-HT_2A receptors and that the serotonergic input onto these cells could influence olfactory functioning at all postnatal ages. © 1995 Wiley-Liss, Inc.     

Involvement of specific placental antigen X-P2 in rat olfaction: an immunohistochemical study in the olfactory bulb

Human placental antigen X-P2 (hPAX-P2), an antigen complex associated with cytochrome P-450 of aromatase within estrogen synthesizing tissues, has been reported to be present in a distinct group of rat primary olfactory receptors involved in suckling behavior. In this study, most of the mitral and tufted cells in the rat olfactory bulb were found to possess hPAX-P2 immunoreactivity. This suggests that the activity of these cells can be hormonally modulated and that hPAX-P2 is involved in rat olfaction not only at the receptor level but also at integrative brain levels via the secondary projecting neurons of the olfactory pathway.     

Localization of tyrosine hydroxylase and olfactory marker protein immunoreactivities in the human and macaque olfactory bulb.

These studies utilized specific antisera to examine the distribution and characteristics of tyrosine hydroxylase and olfactory marker protein in the olfactory bulb of the human and macaque monkey. The macaque displayed immunoreactive profiles to both antisera comparable to those described previously for other mammals. Olfactory marker protein antiserum labeled the olfactory nerve layer and glomeruli. Within the glomeruli, labeled processes were interdigitated with unlabeled processes believed to be the postsynaptic dendrites of olfactory bulb neurons. Tyrosine hydroxylase antisera labeled somata surrounding the glomeruli as well as putative dendritic processes with the glomerular neuropil. It appeared that only a subset of juxtaglomerular neurons were immunoreactive. A similar pattern was observed in the human for both antibodies. Fascicles of olfactory marker protein immunoreactive olfactory nerves often coursed long distances into the olfactory bulb prior to arborizing within a glomerulus. The data suggest that olfactory receptor cell axons destined for specific glomeruli fasciculate into bundles prior to reaching the target glomeruli. The immunoreactivity in the human to tyrosine hydroxylase was qualitatively similar to the macaque and other mammals although the number of labeled somata and intraglomerular processes appeared lower. As in the macaque, it appeared that only a subset of juxtaglomerular neurons were labeled.     

Postmitotic, postmigrational expression of tyrosine hydroxylase in olfactory bulb dopaminergic neurons

The developmental expression of tyrosine hydroxylase (TOH) was studied in a large, specific population of dopaminergic (DA) neurons in the main olfactory bulb (MOB) of the rat. These DA neurons comprise an anatomically distinctive population that has been well characterized in the adult hamster (Davis and Macrides, 1983) and rat (Halasz et al., 1981; Baker et al., 1983, 1984). We addressed a basic question in developmental neurobiology: What factors regulate the expression of neuronal transmitter phenotype during development? Olfactory bulb DA neurons are born in the ventricular and subependymal zones and migrate through all intervening layers to the most superficial layer in the bulb (Altman, 1969; Bayer, 1983). The time of TOH expression in these neurons was determined using immunohistochemistry and light microscopic image-analysis techniques. The results indicate that TOH phenotype is not expressed when the cells are born in the subependymal zone nor during their migration to the periglomerular region but only after they reached their final destination, the glomerular layer. This suggests that epigenetic factors associated with the glomeruli initiate the expression of the key transmitter synthesizing enzyme in these neurons. Primary olfactory neurons in the nasal epithelium project exclusively to glomeruli of the MOB; removal of this input in adult rats (Kawano and Margolis, 1982; Baker et al., 1983, 1984), mice (Nadi et al., 1981; Baker et al., 1983), dogs (Nadi et al., 1981), and hamsters (Kream et al., 1984) appears to down-regulate the expression of the TOH in periglomerular cells. The present results suggested that the input from the primary olfactory nerve is also necessary for the initial expression of the TOH phenotype. In support of this notion, we found that lesions of the olfactory nerve during the first postnatal week caused a significant reduction in the number of TOH-positive juxtaglomerular neurons in the following weeks. Thus, the olfactory nerve appears to be necessary for both the initiation and maintenance of TOH expression in olfactory bulb neurons. These findings suggest that specific cell-cell interactions play a key role in CNS neuronal transmitter phenotype regulation.(ABSTRACT TRUNCATED AT 400 WORDS)