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Adult olfactory ensheathing cell transplantation for acute spinal cord injury.

Cellular transplantation strategies have been explored for the treatment of spinal cord injury. In particular, olfactory nerve ensheathing cell (OEC) transplantation has been reported to improve functional outcome following injury. We investigated the effect of OEC transplantation using cells derived from adult animals on the restoration of function following a contusion injury to the spinal cord. The NYU impactor was used to create a moderate to severe spinal cord injury in 17 rats. Hoescht stained cultured OECs derived from adult rats (n = 7) or culture medium alone (n = 10) were injected into the injury site immediately following injury. Histological and functional outcomes were measured using immunohistochemistry and the Basso, Beattie, and Bresnahan (BBB) scale. All animals transplanted with OECs were found to have surviving Hoescht positive cells within the spinal cord when sacrificed 6 weeks following injury. Immunohistochemical staining of the explanted cords revealed that the surviving cells stained positively for nerve growth factor receptor. Functional outcomes were not different between the transplanted and control groups. OECs transplanted immediately following a contusion injury to the spinal cord survive during the first 6 weeks following injury. These cells do not appear to influence functional outcome during the first 6 weeks following injury. Additional studies are required in order to definitively determine the utility of this type of cellular transplantation for spinal cord injury.

Animals↗

Role of astroglial extracellular matrix in the formation of rat olfactory bulb glomeruli.

We describe the development of the olfactory bulb (OB) glomeruli, focusing on the role of purported astroglial-associated boundary molecules, as potential guidance factors in glomeruli formation. Glomeruli are unusual structures within the OB formed by extremely tight bundles of afferent fibers that fail to grow deeply into the brain. Using olfactory marker protein antibodies, we found that olfactory axons of stage E17-E18 rat embryos were organized into long strands at the outer-most edge of the bulb. By E20-E21 the axons clustered and formed small spheres and by PD1 they gathered into distinct glomeruli. Prior to and during the transformation of olfactory afferents into glomeruli, there was an enhanced expression of cytotactin/tenascin (CT) and chondroitin sulfate-containing proteoglycan (CS-PG) associated with VIM/GFAP-positive astroglial cell processes at the base of the glomeruli. During later stages of development (PD3-PD9), the GFAP-positive astroglial processes beneath and within the base of the glomeruli showed intensified GFAP staining while maintaining expression of CT and CS-PG. Some astroglial processes had extended and others went even further peripherally into the olfactory nerve layer (ONL) to encapsulate and fully infiltrate the glomeruli. By PD9, CS-PG and CT levels increased inside and outside the glomeruli. However, by PD15 both molecules had decreased inside the glomeruli but remained highly concentrated around the glomeruli to form rings. Thus, at later stages, axon/glia interactions which result in astroglial rearrangement and matrix alterations in relation to the glomeruli occur. Western blots of stage E20 bulb proteins revealed the presence of three proteoglycan core proteins each containing chondroitin-6-sulfate moieties. In addition, a type of keratan sulfate (KS) glycosaminoglycan was present in a core protein not associated with chondroitin-6-sulfate. Our data suggest that astroglial-associated ECM molecules CT and CS-PG, and perhaps KS-PG, present in the core of the early developing OB, could form a molecular "wall" that helps confine ingrowing olfactory axons within the ONL at the outer edge of the astroglial territory. We suggest that the astroglia do not precisely regulate the specific shape of the glomeruli, but rather play a fundamental role in directing the gross position at which formation of the synaptic glomeruli will occur.

Aging↗

Hepatocyte growth factor acts as a motogen and guidance signal for gonadotropin hormone-releasing hormone-1 neuronal migration.

Reproduction in mammals is under the control of the hypothalamic neuropeptide gonadotropin hormone-releasing hormone-1 (GnRH-1). GnRH-1-secreting neurons originate during embryonic development in the nasal placode and migrate into the forebrain along olfactory nerves. Gradients of secreted molecules may play a role in this migratory process. In this context, hepatocyte growth factor (HGF) is a potential candidate, because it promotes cell motility in developing brain and has been shown previously to act as a motogen on immortalized GnRH-1 neurons (GN11). In this study, the role of HGF and its receptor Met during development of the GnRH-1 system was examined. GnRH-1 cells express Met during their migration and downregulate its expression once they complete this process. Tissue-type plasminogen activator (tPA), a known HGF activator, is also detected in migratory GnRH-1 neurons. Consistent with in vivo expression, HGF is present in nasal explants, and GnRH-1 neurons express Met. HGF-neutralizing antibody was applied to explants to examine the role of the endogenous growth factor. Migration of GnRH-1 cells and olfactory axon outgrowth were significantly reduced, in line with disruption of a guidance gradient. Exogenous application of HGF to explants increased the distance that GnRH-1 cells migrated, suggesting that HGF also acts as a motogen to GnRH-1 neurons. Functional experiments, performed on organotypic slice cultures, show that creation of an opposing HGF gradient inhibits GnRH-1 neuronal migration. Finally, tPA(-/-):uPA(-/-) (urokinase-type plasminogen activator(-/-)) knock-out mice exhibit strong reduction of the GnRH-1 cell population. Together, these data indicate that HGF signaling via Met receptor influences the development of GnRH-1.

Animals↗

The distribution, relationships to other organelles, and calcium-sequestering ability of smooth endoplasmic reticulum in frog olfactory axons.

Ultrastructural studies of single and serial sections of bullfrog olfactory axons showed that smooth endoplasmic reticulum (SER) tubules, which usually appear as single profiles in cross-sections of axons, are continuous over considerable distances, but that discontinuities do exist. Computer-assisted three-dimensional reconstruction of portions of axons indicated that the SER tubules show considerable variation in the volume of the cisternal space along the tubule, which often follows a tortuous path. Some branching and anastomosing appears to occur, and electron-dense material was present in the cisternal space of some tubules. SER tubules are often bridged to neurofilaments and less often to microtubules. The usual two to three microtubules in the axoplasm form a domain which is characterized by a clear area, or zone of exclusion, around the microtubules. Ultrastructural cytochemistry was used to demonstrate that SER tubules actively sequester Ca. The electron-dense product (calcium oxalate) was uniformly and specifically associated with the SER of axons at both proximal (closest to the perikarya in the olfactory epithelium) and distal (closest to the olfactory lobe of the brain) ends of the olfactory nerve. It is concluded that the primary function of SER tubules in these axons is to serve in the regulation of Ca in the axoplasm, probably to facilitate fast axoplasmic transport, and that a secondary function may be the translocation of material in the cisternal space. The observations are discussed as they may relate to the "microstream" hypothesis of axoplasmic transport, and it is argued that fast transport occurs through the zone of exclusion associated with the microtubule domain(s) of axons.

Animals↗

[Frontal sinus approach to olfactory groove meningiomas].

We report on our experience of the frontal sinus approach for removing olfactory groove meningiomas. Five tumors were operated on, one unilateral, four bilateral. Osteotomy of the anterior wall of the frontal sinus was performed with an oscillating saw without any burr hole. The posterior wall of the sinus was resected and the tumor was attacked along the plane of the anterior skull base. Ethmoidal blood supply was controlled at the initial stage of the operation, allowing avascular tumor debulking. Olfactory nerves, invaded by the tumor, usually cannot be spared. Tumor extensions towards the sella and the optic canals were removed without any brain retraction, nor opening of the sylvian fissure, nor dissection of the carotid arteries. The frontal sinus approach is technically easy to achieve. Osteotomy and reconstruction of the anterior wall of the frontal sinus are rapidly performed. When the frontal sinus is small, image guided surgery allows to delineate precisely its limits and the flap includes the calvarial outer layer, tangenitally cut from one supra-orbital canal to the other. Cosmetic result is perfect. The frontal sinus approach gives access to the medial part of the orbital roofs and to the central anterior skull base from the crista galli to the tuberculum sellae and the anterior clinoids. The frontal sinus approach represents an alternative to conventional craniotomies for tumors developed in the central anterior skull base, especially for olfactory groove meningiomas.

Frontal Sinus↗

Nidus and tasseled cell: distinctive neuronal organization of the main olfactory bulb of the laboratory musk shrew (Suncus murinus).

We revealed the structural features of particular synaptic regions, nidi, and newly found neurons, tasseled cells, in the main olfactory bulb (MOB) of the laboratory musk shrew (Suncus murinus). Nidi were intensely immunoreactive for glutamic acid decarboxylase (GAD) and calbindin D28k (CB), were 30-80 microm in diameter, and were located beneath glomeruli, appearing to make glomerulus-nidus unit-like complexes. In contrast to glomeruli, they contained few or no olfactory nerves. Nidi were distributed throughout the whole MOB and made a distinctive layer, nidal layer. Tasseled cells were located in the mitral cell layer and in the middle of the external plexiform layer (EPL) and extended single primary dendrites to the nidus, where their small tuft-like complicated branches intermingled with processes of perinidal cells surrounding nidi. Primary dendrites of mitral/tufted cells also penetrated nidi but passed to glomeruli. In the outer half of the EPL, columnar structures were seen, where CB- and GAD-positive elements appeared to associate with bundles of cylindrical dendrites of presumed mitral/tufted and tasseled cells. By electron microscopic examinations, nidi were confirmed to be particular synaptic areas where GAD-positive processes made symmetrical synapses to GAD-negative presumed tasseled and mitral/tufted cell dendrites and received asymmetrical synapses from the latter. Retrograde tracings revealed that tasseled cells, in addition to mitral/tufted cells, projected their axons to the lateral olfactory tract, indicating that there were two parallel projection systems in the shrew MOB, which might interact with each other via various types of gamma-aminobutyric acid (GABA)ergic interneurons. The present study clearly showed that the neuronal organization of the shrew MOB was distinctly different from that in rodents.

Animals↗

Isolated congenital anosmia with morphologically normal olfactory bulb in two Iranian families: a new clinical entity?

Congenital total loss of the sense of smell occurs as a part of a syndrome or isolated anosmia. Kallmann syndrome is the most well known congenital anosmia associated with hypogonadotropic hypogonadism. Isolated congenital anosmia (ICA) is a very rare condition and appears to be due to changes in the olfactory epithelium or to aplasia of the olfactory nerve, bulb, and tract. Here we report two unrelated Iranian families with ICA. One family consisted of nine affected members, and the other family contained three affected members. Clinical history, physical examination, and smell testing by intravenous injection of combined vitamins (Alinamin trade mark, Takeda Pharmaceutical Co. Ltd., Japan) confirmed the disease in each affected member. No signs of hypogonadism or other neurological disorders were observed in any affected members. Family analysis with the complete ascertainment method under assumption of the same condition in the two families suggested that the disease is not inconsistent with an autosomal dominant mode with incomplete penetrance. The inheritance in one family appears unusual, i.e., there were no affected individuals in the third generation. When only two upper generations in the family are concerned, the segregation ratio was 0.39 +/- 0.11. Male-to-male transmissions were observed and both sexes were affected in both families. Magnetic resonance imaging (MRI) of the olfactory bulb and sulcus revealed no evidence of morphological changes in all affected members, suggesting that these patients have either a defect in the olfactory epithelium or a functional defect in the olfactory cortex.

Female↗

Glycogen phosphorylase activity in the olfactory bulb of the young rat.

The activity of glycogen phosphorylase, the enzyme that controls glycogen breakdown, was histochemically mapped in the olfactory bulbs of 19-day-old rats. The effect of early odor experience on subsequent olfactory bulb phosphorylase activity was also examined. The highest level of phosphorylase staining in the bulb (and seemingly the highest in the brain) was in the glomerular layer, followed by the external plexiform, internal plexiform, granule cell, and olfactory nerve layers. Virtually no activity was visible in the large output neurons of the bulb, mitral, and tufted cells. Early peppermint odor experience, previously shown to increase metabolic activity in specific glomerular foci as measured by 2-deoxyglucose uptake, had no apparent effect on glomerular-layer phosphorylase activity. In some odor-familiar animals, however, patches of activity were seen in the internal plexiform layer in the area of the bulb where foci of high deoxyglucose uptake are seen in response to peppermint. The patches were directly in line with modified glomerular clusters often seen to underlie foci of enhanced deoxyglucose uptake. The existence of particularly heavy activity in the peripheral third of the glomerular layer, where glycogen-containing modified Schwann cells have been localized, raises the possibility that the glomerular-layer activity is at least partially glial in origin. Finally, because of its rich noradrenaline and serotonin innervation and high density of insulin receptors, the olfactory bulb is proposed as a model system to study the interaction of glycogen/glucose metabolism with neural activity in a relatively well-defined neuronal circuit.

Age Factors↗

Canine distemper virus uses both the anterograde and the hematogenous pathway for neuroinvasion.

Canine distemper virus (CDV), a member of the Morbillivirus genus that also includes measles virus, frequently causes neurologic complications, but the routes and timing of CDV invasion of the central nervous system (CNS) are poorly understood. To characterize these events, we cloned and sequenced the genome of a neurovirulent CDV (strain A75/17) and produced an infectious cDNA that expresses the green fluorescent protein. This virus fully retained its virulence in ferrets: the course and signs of disease were equivalent to those of the parental isolate. We observed CNS invasion through two distinct pathways: anterogradely via the olfactory nerve and hematogenously through the choroid plexus and cerebral blood vessels. CNS invasion only occurred after massive infection of the lymphatic system and spread to the epithelial cells throughout the body. While at early time points, mostly immune and endothelial cells were infected, the virus later spread to glial cells and neurons. Together, the results suggest similarities in the timing, target cells, and CNS invasion routes of CDV, members of the Morbillivirus genus, and even other neurovirulent paramyxoviruses like Nipah and mumps viruses.

Animals↗

Correlation between insulin-like growth factor (IGF)-binding protein 5 and IGF-I gene expression during brain development.

Insulin-like growth factor (IGF)-binding proteins (IGFBPs) potently modulate the interactions of IGF-I and -II with the IGF-I receptor. Previous studies have shown that IGFBP2 gene expression is localized in astroglia, where it is anatomically and temporally coordinated with neuronal IGF-I expression during postnatal brain development. The present study shows that IGFBP5 gene expression is also highly abundant during brain development and also demonstrates significant spatiotemporal correlation with IGF-I, but exhibits a neuroanatomical distribution that is entirely distinct from IGFBP2. IGFBP5 and IGF-I mRNAs are synchronously coexpressed in principal neurons of sensory relay systems, including the olfactory bulb, medial and dorsal lateral geniculate bodies, and ventral tier, cochlear, lemniscal, and vestibular nuclei. They are also transiently coexpressed in principal neurons of the anterodorsal nucleus, but IGF-I mRNA disappears from this structure shortly after birth, while IGFBP5 mRNA remains highly abundant here in the adult. IGFBP5 and IGF-I gene expression demonstrate a temporally coordinated laminar association in the developing cerebellar cortex and hippocampal formation. IGF-I mRNA is concentrated in Purkinje cells, while IGFBP5 mRNA is localized in the external germinal zone in the developing cerebellar cortex. IGFBP5 mRNA is transiently expressed in the retrosplenial and cingulate cortex, subiculum, Ammon's horn, and amygdala, while IGF-I mRNA is contemporaneously localized in large interneurons distributed throughout the hippocampal formation. IGFBP5 mRNA is localized in the lateral ventricular germinal zone at birth and remains in the subventricular zone into maturity. It is also detected in forebrain white matter tracts and olfactory nerve from the second week after birth into maturity. Thus IGFBP5, in addition to IGFBP2, may be a significant determinant of IGF action in the brain. Colocalization in some sites and paralocalization with IGF-I in other sites suggests the potential for autocrine and paracrine interaction between the binding protein and IGF-I in different settings. Arguments are advanced suggesting a role for IGFBPs in the targeting of IGF action to specific cell addresses during brain development.

Animals↗

Olfactory evoked responses and identification tests in neurological disease.

UNLABELLED: To assess the value of smell testing we used olfactory evoked potentials (OEP) and an identification test in multiple sclerosis, Parkinson's disease, motor neuron disease and Alzheimer's disease. METHODS: The OEP to H2S (20 ppm) was obtained using an olfactometer designed to stimulate olfactory nerve endings only. Odor recognition was assessed by the University of Pennsylvania Smell Identification Test (UPSIT). In all instances the disease was 'definite' based on standard diagnostic criteria. Controls were derived from 156 healthy people. RESULTS: 1) Multiple Sclerosis: 11/72 patients (15%) were abnormal on UPSIT. For OEP there was significant increase of latency and decrease in amplitude in 6/26 patients (23%). 2) Parkinson's Disease: 126/155 (81%) patients had an abnormal UPSIT score. 12/37 (32%) had prolonged latency with normal amplitude measurement on OEP, but 27 had absent or unclear readings. 4/10 with normal UPSIT displayed abnormality on OEP. 3) Motor Neuron Disease: 9/58 (16%) were abnormal on UPSIT. There was significant delay in 1/10 (10%) patients on OEP. 4) Alzheimer's Disease: UPSIT scores were abnormal in all 8 patients examined. OEP was normal in 4 of these who could be tested. CONCLUSION: Smell dysfunction was found in all 4 conditions but most severely in Parkinson's Disease (over 80%). The UPSIT in general showed abnormality more frequently than OEP. The olfactory defect probably involves peripheral structures in all diseases tested except Alzheimer's. A patient with normal olfaction is unlikely to have idiopathic Parkinson's disease.

Adolescent↗

Voltage imaging from dendrites of mitral cells: EPSP attenuation and spike trigger zones.

To obtain a more complete description of individual neurons, it is necessary to complement the electrical patch pipette measurements with technologies that permit a massive parallel recording from many sites on neuronal processes. This can be achieved by using voltage imaging with intracellular dyes. With this approach, we investigated the functional structure of a mitral cell, the principal output neuron in the rat olfactory bulb. The most significant finding concerns the characteristics of EPSPs at the synaptic sites and surprisingly small attenuation along the trunk of the primary dendrite. Also, the experiments were performed to determine the number, location, and stability of spike trigger zones, the excitability of terminal dendritic branches, and the pattern and nature of spike initiation and propagation in the primary and secondary dendrites. The results show that optical data can be used to deduce the amplitude and shape of the EPSPs evoked by olfactory nerve stimulation at the site of origin (glomerular tuft) and to determine its attenuation along the entire length of the primary dendrite. This attenuation corresponds to an unusually large mean apparent "length constant" of the primary dendrite. Furthermore, the images of spike trigger zones showed that an action potential can be initiated in three different compartments of the mitral cell: the soma-axon region, the primary dendrite trunk, and the terminal dendritic tuft, which appears to be fully excitable. Finally, secondary dendrites clearly support the active propagation of action potentials.

Action Potentials↗

Optical monitoring of activity from many areas of the in vitro and in vivo salamander olfactory bulb: a new method for studying functional organization in the vertebrate central nervous system.

We have investigated the use of voltage-sensitive dyes to monitor neuronal activity in the intact salamander olfactory bulb. After a 10- to 20-min staining period, a magnified image of an in vitro or an in vivo preparation was formed on a 124-element photodiode array. The array was used to simultaneously record absorption or fluorescence changes from 124 adjacent areas of the bulb. At the magnifications used, each detector received light from 100 to 1000 neurons. Relatively large absorption and fluorescence signals were found in response to olfactory nerve stimulation; all of the results presented were from single trials. Because of the large signal size, measurements on in vivo preparations using epi-illumination also had good signal-to-noise ratios. There were significant differences in signal time course between adjacent detectors which suggested a spatial resolution on the order of 200 microns. Tentative assignments of the cellular origins of some signals could be made from the results of paired volley experiments. The results suggest that optical monitoring of membrane potential could provide a useful method for studying neuronal organization in the intact vertebrate central nervous system.

Ambystoma↗

Response characteristics of an identified, sexually dimorphic olfactory glomerulus.

Partitioning of synaptic neuropil into glomeruli is a common feature of primary olfactory centers in most animal species. The functional significance of glomeruli, however, is not yet well understood. The present study is part of our effort to test the hypothesis that each glomerulus is a functional unit dedicated to processing information about a particular odorant or attribute of odor molecules and that the glomerular array constitutes a map of "odor space." We investigated the physiological and morphological features of uniglomerular projection neurons (PNs) associated with an identified glomerulus in each antennal lobe of the female sphinx moth, Manduca sexta. This "lateral large female glomerulus" (latLFG) is sexually dimorphic and therefore may play a female-specific role, such as processing of information about one or more odorants important for orientation of a female to host plants for oviposition. Together with the medial LFG (medLFG), the latLFG resides outside the array of spheroidal ordinary glomeruli, near the entrance of the antennal (olfactory) nerve. Each LFG is innervated by four to five PNs. Using intracellular recording and staining, we examined the responses of latLFG-PNs to odorants that represent major classes of volatiles released by host plants of M. sexta. All latLFG-PNs were excited when the ipsilateral antenna was stimulated with low concentrations of the monoterpenoid linalool. Dose-response analysis showed that neither other monoterpenoids nor representatives of other classes of host plant volatiles were similarly stimulatory to latLFG-PNs. These findings are consistent with the idea that each glomerulus has a characteristic, limited molecular receptive range.

Action Potentials↗

Neuronal gap junctions in the rat main olfactory bulb, with special reference to intraglomerular gap junctions.

The structural features of neuronal gap junction-forming processes in the rat olfactory bulb were analyzed electron microscopically. Gap junctions were present in glomeruli and extraglomerular regions. In extraglomerular regions, mitral/tufted cell somata, dendrites and axon hillock-initial segments made gap junctions and mixed synapses with interneuronal processes, some of which were confirmed to be GABA positive. In glomeruli gap junctions were encountered mainly between mitral/tufted cell dendrites and diverse types of processes; a small population of them were conclusively identified as periglomerular cell dendrites. Gap junction-forming processes frequently received synapses from olfactory nerve terminals, suggesting that they could be type 1 periglomerular cells. However, the majority were GABA negative or only faintly positive and none were tyrosine hydroxylase positive, indicating that they were different from previously reported type 1 periglomerular cells. Furthermore serial sectioning analyses revealed that the majority of those processes forming gap junctions with mitral/tufted dendrites were smooth cylindrical and had few presynaptic sites, indicating that they were different from previously described periglomerular cells. These findings revealed that mitral/tufted cells make gap junctions with diverse types of neurons; and some of these gap junction-forming processes originated from some types of periglomerular cells but others from hitherto uncharacterized neuron type(s).

Animals↗

A light and electron microscopic study of taurine-like immunoreactivity in the main olfactory bulb of frogs.

The distribution of taurine in the frog olfactory bulb was studied using light and electron microscopic immunohistochemical techniques. At the light microscopic level, taurine-like immunoreactivity (taurine-LI) was found in (i) fibers coursing from the olfactory nerve layer to the glomerular layer, (ii) cell bodies and processes primarily located in the caudal part of the granule cell layer (GCL), and (iii) puncta outlining unstained somata of mitral cells and cells in the GCL. In consecutive sections processed for taurine or GABA, numerous cells of the caudal GCL displayed taurine-LI and GABA-like immunoreactivity (GABA-LI). A bimodal distribution of the cross-sectional cell area for GABA-LI cells implied their morphological diversity, and the peak for larger GABA-LI cells coincided with the maximum for taurine-LI cells. At the electron microscopic level, single immunogold labeling showed that GABA-LI, but not taurine-LI, is present in granule cells, whereas both taurine-LI and GABA-LI were localized in a 'non-granule' type of cell. The double labeling procedure demonstrated coexistence of taurine-LI and GABA-LI in neurons of a 'non-granule' type. These cells had some ultrastructural features typical of short axon cells in the GCL of the mammalian olfactory bulb and were tentatively considered as short axon-like cells. Results suggest that, in the frog olfactory bulb, taurine is contained in primary olfactory afferents and short axon-like cells of the GCL co-localizing GABA and taurine.

Animals↗

Optical recording of odor-evoked responses in the olfactory brain of the naïve and aversively trained terrestrial snails.

Regular spontaneous oscillations were recorded both electro- and optophysiologically using a voltage-sensitive absorption dye in the olfactory part of the brain (procerebral lobe of the cerebral ganglia) of the gastropod mollusk Helix lucorum. Odor application caused transient changes in procerebral oscillations, and an odor-evoked potential was recorded in the procerebrum (PC). The wave of evoked potential originated near the place of olfactory nerve entrance into the PC and propagated via the procerebral neuropile toward the cell body layer. The spread of the odor-evoked potential corresponded roughly to the neuropile area, whereas the spontaneous oscillations were recorded in the cell body layer of the PC and were not observed in the neuropile. Evoked potential did not produce additional events intercalated into the ongoing spontaneous oscillations. Changes in parameters of spontaneous oscillations to the repeated presentations of the same odor were variable. To estimate the role of spontaneous oscillations in odor encoding, we trained the snail to avoid cineole, using paired presentations of cineole and electric shock. Elaboration of conditioned aversion to cineole applications resulted in distinct pairing-specific changes in behavior of the snails and procerebral activity. Responses to odor (cineole) applications were not different in amplitude or frequency of spontaneous oscillations in control and trained snails, whereas ratio of amplitudes of the same oscillation wave in proximal and distal regions of the procerebrum was significantly different in control and aversively trained snails, reflecting changes in neural firing in certain areas of the olfactory lobe.

Animals↗

Properties of external plexiform layer interneurons in mouse olfactory bulb slices.

In the external plexiform layer (EPL) of the main olfactory bulb, apical dendrites of inhibitory granule cells form large numbers of synapses with mitral and tufted (M/T) cells, which regulate the spread of activity along the M/T cell dendrites. The EPL also contains intrinsic interneurons, the functions of which are unknown. In the present study, recordings were obtained from cell bodies in the EPL of mouse olfactory bulb slices. Biocytin-filling confirmed that the recorded cells included interneurons, tufted cells, and astrocytes. The interneurons had fine, varicose dendrites, and those located superficially bridged the EPL space below several adjacent glomeruli. Interneuron activity was characterized by high frequency spontaneous excitatory postsynaptic potential/currents that were blocked by the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione and largely eliminated by the voltage-sensitive Na+ channel blocker, tetrodotoxin. Interneuron activity differed markedly from that of tufted cells, which usually exhibited spontaneous action potential bursts. The interneurons produced few action potentials spontaneously, but often produced them in response to depolarization and/or olfactory nerve (ON) stimulation. The responses to depolarization resembled responses of late- and fast-spiking interneurons found in other cortical regions. The latency and variability of the ON-evoked responses were indicative of polysynaptic input. Interneurons expressing green fluorescent protein under control of the mouse glutamic acid decarboxylase 65 promoter exhibited identical properties, providing evidence that the EPL interneurons are GABAergic. Together, these results suggest that EPL interneurons are excited by M/T cells via AMPA/kainate receptors and may in turn inhibit M/T cells within spatial domains that are topographically related to several adjacent glomeruli.

Action Potentials↗