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Catecholamine innervation of the basal forebrain. III. Olfactory bulb, anterior olfactory nuclei, olfactory tubercle and piriform cortex.

The catecholamine innervation of the olfactory bulb, anterior olfactory nuclei, olfactory tubercle and piriform cortex was studied in the rat using biochemical analysis and fluorescence histochemistry. Biochemical studies demonstrate a moderate norepinephrine (NE) content in all olfactory structures, a high dopamine (DA) content in the olfactory tubercle and a low DA content in the olfactory bulb, anterior olfactory nucleus and piriform cortex. Following locus coeruleus lesions NE content decreases 71% in the olfactory bulb, 82% in the anterior olfactory nucleus, 62% in olfactory tubercle and 77% in piriform cortex...

Amygdala

Mechanisms of augmented field potential responses in the rat olfactory bulb.

Olfactory bulb field potential and single unit responses to paired pulse stimulation of the lateral olfactory tract were observed in the rat. In agreement with previous reports, surgical levels of barbiturate anesthesia prolonged the period of diminution of the field potential responses to the test pulse relative to the effect seen under light anesthesia. Very deep barbiturate anesthesia (sufficient to depress the EEG) led to decreased responses to the primary pulse but augmented responses to the test pulse. Urethane and ether anesthesia produced augmented test responses without severely depressing the EEG and without producing a period of diminished test responses. Single unit recordings failed to show external plexiform layer units excited at the time of maximal augmentation of the field potential. Simultaneous single unit and field potential recordings showed that prolonged diminution of the field potential response was associated with prolonged suppression of unit responses to the test pulse, while augmented field potential responses were associated with decreased suppression of unit responses to the test pulse. These observations are most easily explained by the assumption of temporal facilitation in the synaptic output of mitral and tufted cells. This facilitation is masked by feedback inhibition under some conditions, particularly under barbiturate anesthesia.

Anesthesia, General

Efferent projections of the main and the accessory olfactory bulb in the tree shrew (Tupaia glis).

The projections of the main and the accessory olfactory bulb in the tree shrew (Tupaia glis) have been analyzed with anterograde degeneration and autoradiographic methods for identifying axonal projections, and with the horseradish peroxidase method for identifying the distribution of neurons from which these projections originate. The cytoarchitectonic features of the paleocortical areas which receive projections from the main and the accessory olfactory bulb have also been described. The efferent projections of the accessory olfactory bulb are distributed to the bed nucleus of the accessory olfactory tract, the medial amygdaloid area, the posteromedial cortical amygdaloid area, and to the caudal portion of the bed nucleus of the stria terminalis. In contrast, the efferent projections of the main olfactory bulb are distributed to the anterior olfactory nucleus, the tenia tecta, the olfactory tubercle, the pyriform cortex, the anterior cortical amygdaloid area, the posterolateral cortical amygdaloid area, and to the lateral entorhinal cortex. These observations are consistent with the notion that the olfactory system can be divided into at least two major subsystems: one related to the vomeronasal organ and accessory olfactory bulb, and another related to the main olfactory organ and main olfactory bulb. The paleocortical areas receiving olfactory projections have three basic layers: a superficially positioned plexiform layer (layer I), a pyramidal cell layer (layer II), and a polymorphic cell layer (layer III). The projections of both the main and the accessory olfactory bulb terminate in the outer portion of the plexiform layer (sublamina Ia). Sublamina Ia contains the distal segments of dendrites which originate from a heterogeneous population of neurons located in layer II and, to a lesser extent, layer III. Although the efferent projections of the main and the accessory olfactory bulb are segregated, evidence for a more refined topographical organization within these projections was not obtained. However, the distribution of retrogradely labeled neurons in the main olfactory bulb, following injections of horseradish peroxidase into its various paleocortical targets, indicates that the olfactory projections to these areas may not all originate from the same population of cells.

Amygdala

The locus and cytoarchitecture of the projection areas of the olfactory bulb in Macaca mulatta.

A study was made of the normal and experimental anatomy of the olfactory system of the young adult male rhesus monkey. The cytoarchitecture of the central olfactory areas was studied with cell and fiber stains, while the extent and pattern of the projections of the olfactory bulb were determined by the Fink-Heimer and autoradiographic methods. The brain of one animal that had sustained damage to the olfactory bulb two days prior to sacrifice, and of one that had a transection of the olfactory tract ten days prior to sacrifice, were processed with the Fink-Heimer technique. The first of these and four others received injections of 3H-proline or 3H-leucine into the olfactory bulb, and following a survival period of 18 hours, or 2, 4, 12, or 20 days, their brains were processed with the autoradiographic technique. The results were the same for both experimental methods and for all survival periods. The projections of the olfactory bulb in this microsmatic animal are entirely ipsilateral. All of the structures that receive direct olfactory afferents have a laminar organization except for the anterior olfactory nucleus, which is laminated only in its anterior, peduncular, portion. While the olfactory bulb projects to the entire extent and depth of the anterior olfactory nucleus, the olfactory afferents of all other structures are confined to layer IA of the plexiform layer. These structures are: all divisions of the olfactory tubercle; the frontal and temporal prepiriform cortices; the oral, medial, and dorsal divisions of the superficial amygdaloid nucleus; and polar and anterior entorhinal cortex. The rhesus monkey does not have a recognizable accessory olfactory bulb, and no projections were seen to one of its targets, the nucleus of the stria terminalis. Also, no projections were seen to the taenia tecta or the ventral division of the superficial amygdaloid nucleus. With these exceptions, the projections of the olfactory bulb in the rhesus monkey are similar to those in macrosmatic species.

Afferent Pathways

The postnatal development of the main olfactory bulb of the rat.

The postnatal development from birth to 1 year of the main olfactory bulb was examined quantitatively. The volume of the main olfactory bulb increased over seven-fold by day 30 and remained unchanged thereafter. During the same period the volume of the granular layer increased 18-fold and the mean areas of the olfactory glomeruli increased seven-fold. The mean areas of mitral cell perikarya doubled between the neonatal and juvenile periods. The total number of the mitral cells, however, declined during the first three postnatal weeks. In the internal granular layer of the main olfactory bulb, 89% of the granule cells were acquired postnatally. Much of the cellular gain occurred during the first 3 weeks, with the period of maximum acquisition between days 8 and 14. The number of subependymal cells, the precursors of granule cells, reached a peak at 12 days and gradually declined. But some primitive cells could still be found at one year of age and there was an increase in the total number of granule cells beyond day 30. The mean nuber of internal granular layer cells in a single main olfactory bulb of adult rats was about 5 X 10(6); the number of mitral cells about 4 X 10(4). In the animals injected with 3H-thymidine on day 20 and killed 2 h after injection a small but significant proportion of cells was labelled in the subependymal layer but few in the internal granular layer. In the animals killed 20 and 40 days after injection there was a 10--11-fold rise in the proportion of labelled internal granular layer cells. The proportion of labelled internal granular layer cells decreased in longer survival groups but the total number of labelled cells remained the same, even in year-old animals. However, the total number of internal granular layer cells in the sections examined increased with age.

Aging

Unilateral lesions of the olfactory tubercle modifying general arousal effects in the rat olfactory bulb.

The centrifugal control exerted by different arousal states on the rat olfactory bulb was investigated. The olfactory tubercle was unilaterally coagulated with either 1 mA or 3 mA current. The vigilance state parameters and multiunit mitral cell activity were recorded in freely moving rats, stimulated either by their usual food odor or by isoamyl acetate, in a hungry or a satiated state. In each animal, a unilateral lesion affected resting activity and the relative proportion of positive (excitatory) and negative (inhibitory) responses in the same way in both olfactory bulbs; these effects were proportional to the extent of the lesion. In wakefulness, a nutritional modulation of the bulb responses for food odor existed in spite of a slight decrease in the general arousal level. In slow wave sleep (SWS), when compared to a control group, the rats with lesions showed an increase of neocortical desynchronization induced by olfactory stimulation, and a large decrease of mitral cell excitability. Inhibition of the olfactory input, which normally develops during SWS, could involve mainly mesencephalic neurons reaching the olfactory tubercle and the bulb via the ventral part of the medial forebrain bundle.

Acetates

Changes in sensitivity of mice to anticonvulsant drugs following bilateral olfactory bulb ablations.

Changes in sensitivity to anticonvulsant drugs were investigated after bilateral olfactory bulb ablations in mice. The sensitivity to benzodiazepines and acetazolamide increased, whereas that to phenylacetylurea and dipropylacetic acid decreased, and sensitivity to phenobarbital, diphenylhydantoin and trimethadion was not significantly changes after olfactory bulb ablations. Increase in sensitivity to benzodiazepines was the most significant in both electroshock and pentetrazol convulsions. It was suggested that altered activities and denervation supersensitivity in the limbic system, hypothalamus and midbrain might account for these changes in sensitivity to anticonvulsant drugs after olfactory bulb ablations.

Acetazolamide

Water response of the frog olfactory epithelium as observed from the olfactory bulb.

The water response elicited by application of distilled water on the olfactory epithelium was recorded extracellularly from single olfactory bulb neurones. Characteristics of the water response in the frog olfactory epithelium were examined in comparison with those of the water response in the gustatory and palatal organs. 1. Effects of various electrolyte solutions on the generation of the water response were studied by dripping distilled water on the olfactory epithelium after adaptation to each of these electrolyte solutions. Number of the olfactory bulb cells responding to distilled water increased with increasing the charge of the adapting cations and also with decreasing the size of the cations with a few exceptions. 2. Magnitude of the 'water response' increased with decreasing concentration of salt in the solution which was dripped after adaptation to the isotonic solution of the same salt. 3. The water response was effectively depressed by an electrolyte solution but not by a non-electrolyte solution. An electrolyte also depressed effectively the water response which was produced after adaptation to an organic salt solution. 4. The water response was blocked by treatment of the olfactory epithelium with the uranyl ions which had high affinity for phospholipids. A tentative hypothesis on the generating mechanism of the water response in the frog olfactory epithelium is presented on the basis of the present experimental results and the water responses of the gustatory and palatal organs so far reported.

Action Potentials

[A simple operation method on removal of olfactory bulbs of small rodents (author's transl)].

A simple operation method was described for removing the olfactory bulbs of small rodents. The procedure was as follows: 1. Animals were anesthetized by intraperitoneal injection of pentobarbital sodium. 2. The hair of the head was clipped (Fig. 9). 3. The scalp was cut along the midline between the eyes (Fig. 10). 4. A hole was drilled in the skull overlying the olfactory bulbs (Fig, 14, 15). 5. The olfactory bulbs were aspirated with a glass pipette (Fig. 18). 6. After removing the bulbs by suction, the skin was closed by a single suture (Fig. 21).

Animals

Chronic ethanol exposure alters the expression of genes associated with GPCR-related signaling in the olfactory bulb of male mice.

Chronic ethanol exposure, a key feature of alcohol use disorder (AUD), can affect the nervous system, but its molecular impact on the olfactory bulb remains unclear. In this study, an intermittent two-bottle voluntary drinking model was established in male mice, and transcriptome sequencing was performed on olfactory bulb tissues. DESeq2 analysis identified 188 differentially expressed genes, including 68 upregulated and 120 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome pathway database (Reactome) analyses indicated that ethanol-responsive genes were predominantly enriched in receptor-mediated signaling pathways, particularly those linked to G protein-coupled receptor (GPCR) signaling. Protein-protein interaction analysis further identified eight core GPCR-related genes. Quantitative real-time PCR (qRT-PCR) validation revealed that Cxcl10, Grp, Pcp2, and Pdyn were markedly downregulated in the ethanol group. These results suggest that chronic ethanol exposure is associated with transcriptional alterations in the male mouse olfactory bulb and may selectively affect several GPCR-related signaling components. This study provides candidate molecular evidence for further investigation of ethanol-associated olfactory dysfunction.

Chronic ethanol exposure

Ligand binding studies in the mouse olfactory bulb: identification and characterization of a L-[3H]carnosine binding site.

Binding sites for the dipeptide L-carnosine (beta-alanyl-L-histidine) have been detected in membranes prepared from mouse olfactory bulbs. The binding of L-[3H]-carnosine was saturable, reversible and stereospecific and had a Kd of about 770 nM. The stereospecific binding of L-carnosine represented about 30% of the total binding at pH 6.8, and decreased markedly with increasing pH. Binding was stimulated by calcium, unaffected by zinc, magnesium or manganese and inhibited by sodium and potassium. Carnosine binding was sensitive to trypsin and phospholipases A and C, but not to neuraminidase. Nystatin and filipin, which interact with membrane lipids, also interferred with binding. Some peptide analogues of carnosine were potent inhibitors of binding, but a variety of drugs serving as potent inhibitors in other binding systems had no effect on carnosine binding. Carnosine binding to mouse olfactory bulb membranes was 15-fold higher than that seen in membranes prepared from cerebral hemispheres, 5-fold higher than that seen in membranes prepared from cerebral hemispheres, 5-fold higher than in cerebellum membranes and 3-fold higher than in membranes from spinal medulla and the olfactory tubercle-lateral olfactory tract area. Binding sites for 6 other radiolabeled receptor ligands were also detected in bulb membranes. Peripheral deafferentation of the olfactory bulbs by intranasal irrigation with ZnSO4 led to a loss greater than 90% of the L-[3H]carnosine binding in 4--5 days with much smaller losses in binding of the other 6 ligands over a 180-day observation period. This initial loss of carnosine binding after denervation was due to a loss of binding site stereo-specificity followed by a loss of binding sites. The characteristics of the carnosine binding site in olfactory bulb fulfil 6 of the 7 criteria considered relevant for a functional receptor.

Animals

Transmitter histochemistry of the rat olfactory bulb III. Autoradiographic localization of [3H]GABA.

The distribution of [3H]gamma-aminobutyric acid (GABA) labeled elements in rat olfactory bulb was studied by light and electron microscopic autoradiography. [3H]GABA was strongly taken up into glial cells and pericytes in all layers of the bulb. The neuronal uptake of [3H]GABA was mainly seen in certain types of nerve terminals. About one-third of the granule dendritic terminals, some nerve endings of short axon cells, and certain nerve endings of extrabulbar origin showed a strong labeling. Labeling was seen in a small population of the periglomerular, short axon and granule cell bodies. Most cell bodies of these 3 types as well as the mitral cells did not, however, accumulate any appreciable amo9nt of [3H]GABA. The labeling pattern seen after injection of [3H]glycine and [3H]leucine was clearly different from the pattern seen after [3H]GABA injection. The labeling was more uniformly distributed over the components of the neuropil with a considerably higher activity over certain cell somata such as the mitral cells. The present results demonstrate that neuronal uptake and accumulation of [3H]GABA occur into populations of olfactory bulb cells and processes, which from neurophysiological and/or immunohistochemical studies are supposed to use GABA as a neurotransmitter.

Aminooxyacetic Acid

Influence of the pineal gland, olfactory bulbs and photoperiod on surges of plasma prolactin in the female rat.

The role of the pineal gland, olfactory bulbs and photoperiod in the regulation of the two daily surges of plasma prolactin in the pseudopregnant rat has been investigated. Pinealectomy had no effect on the surges of prolactin in pseudopregnant rats maintained on either a long (14 h light : 10 h darkness; 14L : 10D) or a short (2L : 22D) photoperiod, but olfactory bulbectomy decreased the nocturnal surge in animals maintained on 14L : 10D. This effect of bulbectomy was eliminated if the rats maintained on 14L : 10D were also pinealectomized. After cervical stimulation, bulbectomized rats maintained on a 2L : 22D photoperiod had nocturnal-type prolactin surges similar to those of intact rats maintained on the same photoperiod. These results indicate that the pineal gland andlength of photoperiod are not involved in the regulation of the surges of plasma prolactin in pseudopregnant rats but that the olfactory bulbs may enhance the nocturnal surge.

Animals