Mitral Cell Layer Research Articles

Morphological and quantitative features of mitral cells in the rat olfactory bulb after administration of propranolol.

To determine the cytoplasmic morphological changes in the mitral cells and quantitative changes (number of mitral cells and thickness of mitral cell layer in microns) in the rat olfactory bulb after administration of propranolol. Experimental study. The Department of Anatomy, University of Health Sciences, Lahore, from January 2006 to January 2007. Twenty samples were obtained from two randomly divided groups of rats, each comprising 10 animals for control and experimental work respectively. Each group was treated with normal saline (5 ml/kg) and propranolol (1 mg/kg) respectively for one month. The skull was fixed in 20% formalin for 10 days and decalcified in 10% formalin/10% nitric acid. The olfactory bulb along with olfactory cortex was dissected. After processing, 10 microns thick sections were obtained. The slides were stained with Hematoxylin and Eosin and Bielschowksy's silver stain (Glees-Marsland modification) and studied under light microscope. The morphology, quantitative analysis of mitral cell layer and the number of mitral cells were studied in the histological study and compared using t-test with significance at p < 0.05. In the propranolol treated group changes observed in the morphology of the mitral cells included presence of cytoplasmic vacuoles at the periphery of the cells. There was significant increase in the thickness of mitral cell layer and number of the mitral cell in the propranolol treated group (p < 0.05). This study showed morphological and quantitative changes in the olfactory bulb in response to treatment with propranolol, hence it has implications in odour induced learning.

Synaptic Inhibition in the Olfactory Bulb Accelerates Odor Discrimination in Mice

Local inhibitory circuits are thought to shape neuronal information processing in the central nervous system, but it remains unclear how specific properties of inhibitory neuronal interactions translate into behavioral performance. In the olfactory bulb, inhibition of mitral/tufted cells via granule cells may contribute to odor discrimination behavior by refining neuronal representations of odors. Here we show that selective deletion of the AMPA receptor subunit GluA2 in granule cells boosted synaptic Ca(2+) influx, increasing inhibition of mitral cells. On a behavioral level, discrimination of similar odor mixtures was accelerated while leaving learning and memory unaffected. In contrast, selective removal of NMDA receptors in granule cells slowed discrimination of similar odors. Our results demonstrate that inhibition of mitral cells controlled by granule cell glutamate receptors results in fast and accurate discrimination of similar odors. Thus, spatiotemporally defined molecular perturbations of olfactory bulb granule cells directly link stimulus similarity, neuronal processing time, and discrimination behavior to synaptic inhibition.

Characterization of CGRP protein expression in “satellite‐like” cells and dendritic arbours of the mouse olfactory bulb

The main olfactory bulb (OB) is made up of several concentric layers, forming circuitries often involving dendro-dendritic synapses. Important interactions between OB neurons occur in the external plexiform layer (EPL), where dendrites of tufted and Van Gehuchten cells form synapses with dendrites of deeper lying mitral, tufted, and granule cells. OB neurons display a variety of neurotransmitters. Here, the focus is on calcitonin gene-related peptide (CGRP), a 37-amino acid neuropeptide transmitter that is widely distributed in the central and peripheral nervous system. In the OB, CGRP-immunoreactive (ir) cell bodies were mostly observed in the mitral cell layer (MCL) of normal mice, and their number increased following colchicine treatment. Sparsely distributed CGRP-ir cell bodies were also found in the EPL and granular cell layer. Double-immunofluorescence experiments revealed a lack of co-localization between CGRP-like immunoreactivity (LI) and corticotropin-releasing factor- or galanin-LI, two markers for mitral cells, and no CGRP-LI was found in cholecystokinin-, parvalbumin-, or vasoactive intestinal polypeptide-ir tufted/Van Gehuchten cells. CGRP-ir cell bodies were not found to co-localize glutamic acid decarboxylase 67 (GAD67)-green fluorescence protein, gamma-aminobutyric acid (GABA)-, or calretinin-LI, although the possibility remains that CGRP-ir cells may contain low levels of GABA and/or GAD67 not detected by our methodology. Dendrites of CGRP-ir cells extensively ramified deep in the EPL and double-immunofluorescence revealed them to be adjacent with, often apparently contacting, dendrites of granule, mitral, tufted, and Van Gehuchten cells. We propose that these CGRP-ir cell bodies in the mouse OB are "satellite-like" cells within and, occasionally, close to the MCL.

Description of distributed features of the nestin‐containing cells in brains of adult mice: A potential source of neural precursor cells

The distribution of neural precursor cells (NPCs) in adult mice brain has so far not been described. Therefore, we investigated the distribution of NPCs by analyzing the nestin-containing cells (NCCs) in distinct brain regions of adult nestin second-intron enhancer-controlled LacZ reporter transgenic mice through LacZ staining. Results showed that NCCs existed in various regions of adult mouse brain. In cerebellum, the greatest number of NCCs existed in cortex of the simple lobule, followed by cortex of the cerebellar lobule. In olfactory bulb, NCCs were most numerous in the granular cell layer, followed by the mitral cell layer and the internal plexiform, glomerular, and external plexiform layers. In brain nuclei (nu), NCCs were most numerous in the marginal nu, followed by the brainstem and diencephalon nu. NCCs in sensory nu of brainstem were more numerous than in motor nu, and NCCs in the dorsal of sensory nu were more numerous than in the ventral part. In brain ventricle systems, NCCs were largely distributed in the center of and external to the lateral ventricle, the inferior part of the third ventricle, the dorsal and inferior parts of the fourth ventricle, and the gray matter around the cerebral aqueduct. NCCs in the left vs. right brain were not significantly different. These data collectively indicate that NCCs were extensively distributed in the cerebellum and olfactory bulb, the partial nu of the marginal system, the partial brain nu adjacent to brain ventricle systems, the subependymal zone, and the cerebral cortex around the marginal lobe and were a potential source of NPCs.

Down-regulated GABAergic expression in the olfactory bulb layers of the mouse deficient in monoamine oxidase B and administered with amphetamine.

This study explores primarily the role of the activity of monoamine oxidase B (MAOB) in the regulation of glutamic acid decarboxylase(67) (GAD(67)) expression in distinct layers of main olfactory bulb (OlfB), which links the limbic system. Moreover, the response of GAD(67) was investigated to amphetamine perturbation in the absence of MAOB activity. Immunocytochemical analysis was performed on OlfB sections prepared from the adult wild type (WT) and the MAOB gene-knocked-out (KO) mice after receiving repeated intraperitoneal injections (two doses per day, total seven doses) of saline or amphetamine, 5 mg/kg. The levels of the GAD(67) immunoreactivity were approximate 25 and 38% lower in respective glomerular (GloL) and mitral cell layers (ML) of saline-treated KO mice than that of WT, whereas similar in the external plexiform or granule cell layers (GraL) of the KO and WT. In the GloL, the level of tyrosine hydroxylase was 39% lower in the KO mice than WT, implicating different dopamine content in the KO from WT. The amphetamine exposure down-regulated the levels of GAD(67) in the WT layers by 46 to 52%, and in KO layers 65 to 71%, except ML. The GraL GAD(67) level may be regulated by the activation of CREB, as the phosphorylated (p) CREB coexisted with GAD(67), and the percentage of GAD(67)-expressing pCREB neurons was decreased by the amphetamine exposure. The data indicate that the activity of MAOB could modulate the regular and amphetamine-perturbed expression of GAD(67) and pCREB. Thus, interactions are suggested among the MAOB activity, GABA content of OlfB, and olfaction.

Mutation of the Variant α-Tubulin TUBA8 Results in Polymicrogyria with Optic Nerve Hypoplasia

The critical importance of cytoskeletal function for correct neuronal migration during development of the cerebral cortex has been underscored by the identities of germline mutations underlying a number of human neurodevelopmental disorders. The proteins affected include TUBA1A, a major alpha-tubulin isoform, and microtubule-associated components such as doublecortin, and LIS1. Mutations in these genes are associated with the anatomical abnormality lissencephaly, which is believed to reflect failure of neuronal migration. An important recent observation has been the dependence of cortical neuronal migration upon acetylation of alpha-tubulin at lysine 40 by the histone acetyltransferase Elongator complex. Here, we describe a recognizable autosomal recessive syndrome, characterized by generalized polymicrogyria in association with optic nerve hypoplasia (PMGOH). By autozygosity mapping, we show that the molecular basis for this condition is mutation of the TUBA8 gene, encoding a variant alpha-tubulin of unknown function that is not susceptible to the lysine 40 acetylation that regulates microtubule function during cortical neuron migration. Together with the unique expression pattern of TUBA8 within the developing cerebral cortex, these observations suggest a role for this atypical microtubule component in regulating mammalian brain development.

IGF‐I promotes neuronal migration and positioning in the olfactory bulb and the exit of neuroblasts from the subventricular zone

While insulin-like growth factor-I (IGF-I) supports neuronal and glial differentiation in the CNS, it is largely unknown whether IGF-I also influences neuronal migration and positioning. We show here that the pattern of olfactory bulb (OB) layering is altered in Igf-I (-/-) mice. In these animals, Tbr1(+)-glutamatergic neurons are misplaced in the mitral cell layer (ML) and the external plexiform layer (EPL). In addition, there are fewer interneurons in the glomerular layer and the EPL of the Igf-I (-/-) mice, and fewer newborn neurons are incorporated into the OB from the forebrain subventricular zone (SVZ). Indeed, neuroblasts accumulate in the postnatal/adult SVZ of Igf-I (-/-) mice. Significantly, the positioning of Tbr1(+)-cells in a primitive ML is stimulated by IGF-I in cultured embryonic OB slices, an effect that is partially repressed by the phosphoinositide 3-kinase (PI3K) inhibitor. In OB cell cultures, IGF-I increases the phosphorylation of disabled1 (P-Dab1), an adaptor protein that is a target of Src family kinases (SFK) in the reelin signalling pathway, whereas reduced P-Dab1 levels were found in Igf-I (-/-) mice. Neuroblast migration from the rostral migratory stream (RMS) explants of postnatal Igf-I (-/-) was similar to that from Igf-I (+/+) explants. However, cell migration was significantly enhanced by IGF-I added to the explants, an effect that was repressed by PI3K and SFK inhibitors. These findings suggest that IGF-I promotes neuronal positioning in the OB and support a role for IGF-I in stimulating neuroblast exit from the SVZ into the RMS, thereby promoting the incorporation of newly formed neurons into the OB.

Disorganized olfactory bulb lamination in mice deficient for transcription factor AP-2ɛ

Within the olfactory bulb, neurons and their axonal connections are organized into distinct layers corresponding to different functionalities. Here we demonstrate that transcription factor AP-2ɛ is required for olfactory bulb development, specifically the establishment of appropriate neuronal lamination. During normal mouse embryogenesis, AP-2ɛ transcripts are one of the earliest markers of olfactory bulb formation, and expression eventually becomes refined to the projection neurons, the mitral and tufted cells. To assess the function of AP-2ɛ in olfaction, we generated a null allele (the “AK” allele) by inserting a Cre recombinase transgene into the endogenous AP-2ɛ genomic locus. AP-2ɛ-null mice exhibited defective olfactory bulb architecture. The mitral cell layer was disorganized, typified by misoriented and aberrantly positioned projection neurons, and the adjacent internal plexiform layer was absent. Despite the significant disruption of olfactory bulb organization, AP-2ɛ null mice were viable and could discriminate a variety of odors. AP-2ɛ-null mice thus provide compelling evidence for the robust nature of the mouse olfactory system, and serve as a model system to probe both the regulation of neuronal lamination and the functional circuitry of the olfactory bulb. We also show that Cre recombinase expression directed by the AP-2ɛ locus can specifically target floxed genes within the olfactory bulb, extending the utility of this AK allele.

MRI of cellular layers in mouse brain in vivo

Noninvasive imaging of the brain of animal models demands the detection of increasingly smaller structures by in vivo MRI. The purpose of this work was to elucidate the spatial resolution and structural contrast that can be obtained for studying the brain of C57BL/6J mice by optimized T2-weighted fast spin-echo MRI at 9.4 T. As a prerequisite for high-resolution imaging in vivo, motion artifacts were abolished by combining volatile anesthetics and positive pressure ventilation with a specially designed animal bed for fixation. Multiple substructures in the cortex, olfactory bulb, hippocampus, and cerebellum were resolved at 30 to 40 μm in-plane resolution and 200 to 300 μm section thickness as well as for relatively long echo times of 65 to 82 ms. In particular, the approach resulted in the differentiation of up to five cortical layers. In the olfactory bulb the images unraveled the mitral cell layer which has a thickness of mostly single cells. In the hippocampus at least five substructures could be separated. The molecular layer, Purkinje layer, and granular layer of the cerebellum could be clearly differentiated from the white matter. In conclusion, even without the use of a contrast agent, suitable adjustments of a widely available T2-weighted MRI sequence at high field allow for structural MRI of living mice at near single-cell layer resolution.

Nutritional status modulates behavioural and olfactory bulb Fos responses to isoamyl acetate or food odour in rats: roles of orexins and leptin

Food odours are major determinants for food choice, and their detection depends on nutritional status. The effects of different odour stimuli on both behavioural responses (locomotor activity and sniffing) and Fos induction in olfactory bulbs (OB) were studied in satiated or 48-h fasted rats. We focused on two odour stimuli: isoamyl acetate (ISO), as a neutral stimulus either unknown or familiar, and food pellet odour, that were presented to quiet rats during the light phase of the day. We found significant effects of nutritional status and odour stimulus on both behavioural and OB responses. The locomotor activity induced by odour stimuli was always more marked in fasted than in satiated rats, and food odour induced increased sniffing activity only in fasted rats. Fos expression was quantified in periglomerular, mitral and granular OB cell layers. As a new odour, ISO induced a significant increase in Fos expression in all OB layers, similar in fasted and satiated rats. Significant OB responses to familiar odours were only observed in fasted rats. Among the numerous peptides shown to vary after 48 h of fasting, we focused on orexins (for which immunoreactive fibres are present in the OB) and leptin, as a peripheral hormone linked to adiposity, and tested their effects of food odour. The administration of orexin A in satiated animals partially mimicked fasting, since food odour increased OB Fos responses, but did not induce sniffing. The treatment of fasted animals with either an orexin receptors antagonist (ACT-078573) or leptin significantly decreased both locomotor activity, time spent sniffing food odour and OB Fos induction in all cell layers, thus mimicking a satiated status. We conclude that orexins and leptin are some of the factors that can modify behavioural and OB Fos responses to a familiar food odour.

Perisomatic‐targeting granule cells in the mouse olfactory bulb

Inhibitory interneurons in the hippocampus and neocortex are differentiated into several morphological and functional subtypes that innervate distinct subcellular domains of principal neurons. In the olfactory bulb (OB), odor information is processed by local neuronal circuits that include the major inhibitory interneuron, granule cells (GCs). All GCs reported to date target their inhibitory output synapses mainly to dendrites of mitral cells (MCs) and tufted cells (TCs) in the external plexiform layer (EPL). Here we identified a novel type of GC that targets output synapses selectively to the perisomatic region of MCs. In the OB of adult transgenic mice expressing green fluorescent protein (GFP) under the control of nestin gene regulatory regions, we observed cells in the granule cell layer (GCL) that have GC-like morphology and strongly express GFP (referred to as type S cells). Type S cells expressed NeuN and GAD67, molecular markers for GCs. Intracellular labeling of type S cells revealed that their dendrites did not enter the EPL, but formed branches and spines within the GCL, internal plexiform layer, and mitral cell layer. Type S cells typically had huge spines at the ends of the apical dendrites. Some of the terminal spines attached to the perisomatic region of MCs and formed dendrosomatic reciprocal synapses with a presumed granule-to-mitral inhibitory synapse and a mitral-to-granule excitatory synapse. These findings indicate the morphological differentiation of GCs into dendritic-targeting and perisomatic-targeting subsets, and suggest the functional differentiation of the GC subsets in the processing of odor information in the OB.

The organization of feedback projections in a pathway important for processing pheromonal signals

In most of the mammalian sensory systems there are massive cortical feedback projections to early processing stations. The mammalian accessory olfactory system is considered unique in several aspects. It is specialized for processing pheromonal signals and plays a critical role in regulating sociosexual behaviors. Furthermore, pheromonal signals are believed to bypass cortex and reach the hypothalamic behavioral centers after merely three forward projections. Because the organization of the feedback projections in the accessory olfactory system remains largely unclear, the importance of the feedback projections in the processing of pheromonal signals has been ignored. Here we show that in mice the feedback projections from the bed nucleus of stria terminalis (BST) and the vomeronasal amygdala to the accessory olfactory bulb (AOB) are topographically organized and use different neurotransmitters. By retrograde and anterograde tracing, we find that the feedback projection from the BST terminates in the AOB mitral cell layer, whereas that from the amygdala terminates in the AOB granule cell layer. By combining tracing, genetic labeling of GABAergic neurons, and immunostaining against a marker of glutamatergic synapses, we observe that the BST-to-AOB projection is GABAergic whereas the amygdala-to-AOB projection is glutamatergic. In addition, a substantial number of feedback neurons in the amygdala and BST express estrogen receptors. Thus, the accessory olfactory system, like other sensory systems, possesses extensive feedback projections. Moreover, our results suggest that central hormonal cues may modulate the processing of pheromonal signals at early stations through the precisely organized feedback projections.

Postnatal changes in expression of vesicular glutamate transporters in the main olfactory bulb of the rat

Olfactory information is initially processed through intricate synaptic interactions between glutamatergic projection neurons and GABAergic interneurons in the olfactory bulb. Although bulbar neurons and networks have been reported to develop even postnatally, much is yet unknown about the glutamatergic neuron development. To address this issue, we studied the postnatal ontogeny of vesicular glutamate transporters (VGLUT1 and VGLUT2) in the main olfactory bulb of rats, using in situ hybridization, immunohistochemistry, and their combination. In situ hybridization data showed that VGLUT1 mRNA is intensely expressed in differentiating mitral cells and smaller cells of the mitral cell layer (MCL) on postnatal day 1 (P1), and also at lower levels in small- and medium-sized cells, presumably tufted cell populations, of the external plexiform layer (EPL) from P5 onward. VGLUT2 mRNA was expressed in many MCL cell populations on P1, also in small- and medium-sized cells of the EPL at almost the same level as MCL cells between P5 and P7, and became apparently less intense in the MCL than in the EPL from P10 onward. The expression, unlike VGLUT1 mRNA, was also found in small-sized cells of the interglomerular region. In partial agreement with these data, immunohistochemical analyses demonstrated that subsets of mitral and EPL cells are stained for VGLUT1 or VGLUT2, with the former cells coexpressing both subtypes until P5. Moreover, a combined fluorescence in situ hybridization–immunohistochemical dual labeling of the P10 bulb revealed that neither VGLUT1 nor VGLUT2 mRNA is expressed in GABAergic or dopaminergic periglomerular cells, implying their expression in other periglomerular cell subclasses, external tufted cells and/or short-axon cells. Thus, the present study suggests that early in the postnatal development distinct glutamatergic bulbar neurons of rats express spatiotemporally either or both of the two VGLUT subtypes as a specific vesicular transport system, specifically contributing to glutamate-mediated neurobiological events.

An ex vivo preparation of the intact mouse vomeronasal organ and accessory olfactory bulb

The accessory olfactory system (AOS) in mammals detects and processes information from liquid-phase environmental odorants, including pheromones. The AOS carries out tasks such as individual recognition, learning, and decision-making with relatively few stages of neural processing; it thus represents an attractive system for investigating the neural circuits that carry out these functions. Progress in understanding the AOS has long been impeded by its relative inaccessibility to standard physiological approaches. In this report, we detail a novel dissection and tissue perfusion strategy that improves access to the accessory olfactory bulb (AOB) while maintaining afferent connections from sensory neurons in the vomeronasal organ (VNO). Mitral cells demonstrated spontaneous and evoked firing patterns consistent with recent in vivo reports. We assayed cell degradation in the AOB tissue using Fluoro-Jade C and found that the VNO and AOB glomerular, external plexiform, and mitral cell layers showed minimal signs of degeneration for up to 6h. Whereas histology indicated some degeneration in the deep inhibitory granule cell layer over time, electrophysiological assays demonstrated intact inhibitory function on mitral cells. Pharmacological blockade of GABAA receptors with 3μM SR95531 (gabazine) resulted in increased evoked mitral cell activity. Furthermore, mitral cells displayed suppression of responses to preferred urine stimuli when preferred and non-preferred stimuli were mixed, an effect thought to involve functional laterally connected inhibition. These results demonstrate the utility of whole mount ex vivo preparations for studying sensory processing in the AOS, and suggest that similar strategies may improve experimental access to other difficult-to-study neural circuits.

Thrombospondin-1 binds to ApoER2 and VLDL receptor and functions in postnatal neuronal migration

Apolipoprotein E receptor 2 (ApoER2), very low-density lipoprotein receptor (VLDLR), and Dab1 are the main components of the Reelin signalling cascade. Reelin is the sole ligand defined so far in signalling through this pathway. Postnatal migration of neuronal precursors from the subventricular zone (SVZ) to the olfactory bulb (OB), however, depends on ApoER2 and Dab1, but functions independently of Reelin. Here, we show that thrombospondin-1 (THBS-1) is a novel physiological ligand for ApoER2 and VLDLR. THBS-1 is present in the SVZ and along the entire rostral migratory stream (RMS). It binds to ApoER2 and VLDLR and induces phosphorylation of Dab1. In contrast to Reelin, it does not induce Dab1 degradation or Akt phosphorylation, but stabilizes neuronal precursor chains derived from subventricular explants. Lack of THBS-1 results in anatomical abnormalities of the RMS and leads to a reduction of postnatal neuronal precursors entering the OB.

Manganese enhanced MRI reveals functional circuitry in response to odorant stimuli

To investigate the circuitry involved in detecting odorants in the rodent brain, we developed a method using manganese-enhanced MRI (MEMRI) to map the flow of neural information from the olfactory sensory neurons (OSNs) to the central layers of the olfactory bulb. Studies have shown that Mn 2+ enters active neurons and is transported anterogradely to axon terminals where it can cross synapses to functionally trace neural networks. Thus, by delivering MnCl 2 directly into the nasal cavity of mice and then exposing them to defined odorants, Mn 2+ is preferentially taken up by activated OSNs. Using the time course of the MRI signal, we generated maps of Mn 2+ accumulation in the olfactory bulb for both glomerular and mitral cell layers. Results demonstrated that overlapping yet distinct enhancement patterns were produced by exposure to either octanal, acetophenone, or carvone. Notably, areas of Mn 2+ accumulation in the mitral cell layer were similar to those in the glomerular layer consistent with neural information that passes from specific OSNs to specific mitral cells. Finally, by correlating specific Mn 2+ signal peaks to genetically labeled glomeruli that are known to be activated by the odorant octanal, we show that MEMRI maps can be resolved at the level of individual glomeruli.

Developmental regulation of metabotropic glutamate receptor 1 splice variants in olfactory bulb mitral cells

Alternative splicing of the metabotropic glutamate receptor 1 (mGluR1) receptor gene generates two major receptor isoforms, mGluR1a and mGluR1b, differing in intracellular function and distribution. However, little is known on the expression profiles of these variants during development. We examined the mRNA expression profile of mGluR1a/b in microdissected layers and acutely isolated mitral cells in the developing mouse olfactory bulb. This analysis showed that the two mGluR1 variants are differentially regulated within each bulb layer. During the first postnatal week, the mGluR1a isoform replaces GluR1b in the microdissected mitral cell layer (MCL) and in isolated identified mitral cells, coinciding with a developmental epoch of mitral cell dendritic reorganization. Although mGluR1a mRNA is expressed at high levels in both the adult external plexiform layer (EPL) and MCL, Western blotting analysis reveals a marked reduction of the mGluR1a protein in the MCL, where mitral cell bodies are located, and strong labeling in the EPL, which contains mitral cell dendrites. This suggests that there is increased dendritic trafficking efficiency of the receptor in adult. The temporal and spatial shift in mGluR1b/a expression suggests distinct roles of the mGluR1 isoforms, with mGluR1b potentially involved in the early mitral cell maturation and mGluR1a in dendritic and synapse function.

Comparison of Identified Mitral and Tufted Cells in Freely Breathing Rats: II. Odor-Evoked Responses

Mitral and tufted cells are the 2 types of output neurons of the main olfactory bulb. They are located in distinct layers, have distinct projection patterns of their dendrites and axons, and likely have distinct relationships with the intrabulbar inhibitory circuits. They could thus be functionally distinct and process different aspects of olfactory information. To examine this possibility, we compared the odor-evoked responses of identified single units recorded in the mitral cell layer (MCL units), in the core of the external plexiform layer (not at the glomerular border tufted cells), or at the glomerular border of this layer (GB tufted cells) of the entire olfactory bulb. Differences between mitral and tufted cells were observed only when subtle aspects of the responses were explored, such as the firing rate per respiratory cycle or the distribution of firing activity along the respiratory cycle. By contrast, more clear differences were found when the 2 subtypes of tufted cells were examined separately. GB units were significantly more responsive, had significantly higher firing activity, and showed greater activity at the transition between inspiration and expiration. The projection-type tufted cells situated closer to the entrance of the olfactory bulb may thus form a distinct physiological class of output neurons and differ from mitral cells and other tufted cells in the manner of processing olfactory information.

Association of cocaine- and amphetamine-regulated transcript and neuropeptide Y in the forebrain and pituitary of the catfish, Clarias batrachus: A double immunofluorescent labeling study

Cocaine- and amphetamine-regulated transcript (CART) and neuropeptide Y (NPY) are involved in the regulation of food intake, body weight, pituitary hormones, and reproduction. While CART and NPY occupy overlapping fields in the brain of mammals, little is known about the interaction between these peptide-containing systems in other vertebrates. We explored neuroanatomical associations between CART and NPY in the olfactory system, forebrain and pituitary of the catfish, Clarias batrachus, using double immunofluorescence method. NPY-containing fascicles from olfactory receptor neurons innervated the olfactory glomeruli and mitral cell layer in close association with CART-containing terminal fields. Distinct CART- or NPY-containing fibers were seen in the medial olfactory tract. In the dorsal telencephalon, CART- and NPY-immunoreactive axons were closely associated in area dorsalis telencephali/pars lateralis dorsalis (Dld), and posterioris (Dlp). In the ventral telencephalon, while most of the cells of nucleus entopeduncularis (NE) showed the presence of CART as well as NPY, a few cells with only NPY-immunoreactivity were observed. Similarly, a CART and NPY colocalized cell population was prominent in the preoptic area (POA); and a small population of cells with NPY-immunoreactivity was also evident. Other areas where CART and NPY were colocalized included fibers in the tuberal area, inferior lobe, neurohypophysis, proximal pars distalis and pars intermedia of the pituitary. No association between CART and NPY was observed in the thalamus and habenular ganglion. These results suggest that CART- and NPY-peptidergic systems may interact in NE, POA, tuberal area, certain telencephalic areas and pituitary and jointly process information relating to reproduction, feeding and neuroendocrine regulation.

BDNF Promoter-Mediated -Galactosidase Expression in the Olfactory Epithelium and Bulb

The neurotrophin brain-derived neurotrophic factor (BDNF) has been implicated in the generation and differentiation of new olfactory sensory neurons (OSNs) and in the regulation of branching of OSN axons in their target glomeruli. However, previous reports of BDNF mRNA and protein expression in olfactory epithelium and olfactory bulb (OB) have been inconsistent, raising questions on the proposed roles for BDNF. Here, we report on β-galactosidase (β-gal) expression in adult gene-targeted mice where the BDNF promoter drives expression of the Escherichia coli lacZ gene (BDNFlacZneo mice). We find that β-gal is expressed in a small subset of OSNs with axons that reach the olfactory nerve layers throughout the OB. In the OB, we find expression of β-gal in γ-aminobutyric acidergic but not dopaminergic periglomerular cells and external tufted cells and in interneurons located in the mitral cell layer. Our results are inconsistent with the regulation of generation and differentiation of new OSNs elicited by the release of BDNF from horizontal basal cells. The results are consistent with a role for BDNF in competitive branching of OSN axons within the glomeruli of the OB.