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Subclasses of vomeronasal receptor neurons: differential expression of G proteins (Gi alpha 2 and G(o alpha)) and segregated projections to the accessory olfactory bulb.

Differential expression of G proteins (Gi alpha 2 and G(o alpha) and the separate central projections of Gi alpha 2- and G(o alpha)-immunoreactive (ir) vomeronasal receptor neurons were investigated in the mouse and rat using immunocytochemical methods. In the vomeronasal organ (VNO), receptor neurons with their cell bodies located in the middle layer (middle 1/3) of the vomeronasal sensory epithelium express Gi alpha 2. Axons of these Gi alpha 2-ir neurons can be followed from VNO to the anterior part, but not the posterior part, of the nerve-glomerular (N-GL) layer of the accessory olfactory bulb (AOB). Another population of receptor neurons, which are located in the deep layer (basal 1/3) of the vomeronasal sensory epithelium, express G(o alpha), and axons of the G(o alpha)-ir neurons can be traced to the posterior part, but not the anterior part, of the N-GL layers of the AOB. The axons of the two subclasses of receptor neurons are intermingled near the VNO and become segregated as they enter the AOB. Removal of the AOB results in retrograde degeneration of both Gi alpha 2-ir and G(o alpha)-ir receptor neurons in the VNO. These results suggest that at least two subclasses of receptor neurons exist in the VNO: the Gi alpha 2-ir neurons in the middle layer and the G(o alpha)-ir neurons in the deep layer of the VNO. The Gi alpha 2-ir neurons in the middle layer of the VNO project to the anterior part of the AOB, while the G(o alpha)-ir neurons in the deep layer of the VNO project to the posterior half of the AOB. These results are similar to our previous observations in the gray short-tailed opossum, suggesting that the existence of at least two subclasses of receptor neurons in the vomeronasal epithelium with differential projections to the AOB is a conserved feature among mammals.

Animals↗

Characterization of cat nasal afferents and brain stem neurones receiving ethmoidal input.

Stimulation of the nasal mucous membrane can initiate protective reflexes, particularly sneezing and apnea. Very little is known about the receptors in the nasal cavity responsible for initiating these reflexes, which are thought to be mediated by trigeminal rather than olfactory pathways. In the cat, the ethmoidal branch of the ophthalmic division of V carries this afferent information. The objectives of the present study were to determine a set of adequate nasal receptor stimuli capable of initiating these protective reflexes; to characterize the types of information carried in the ethmoidal nerve afferents, by recording from single fibers dissected from the nerve; to determine the conduction velocities of these afferents, by recording from the cell bodies of ethmoidal afferents in the trigeminal ganglion; and to study the second-order neurones in the brain stem on which these afferents make contact. Results of single fiber recording indicated that, in addition to being tactile, approximately half of the neurones studied also responded to noxious chemical stimuli. The conduction velocity of the majority of these afferents were found to be in the A-delta range. Second-order neurones in spinal trigeminal nucleus which had an input from the ethmoidal nerve could be divided into two classifications: (i) low threshold mechanoreceptive (LTM) neurones which received light tactile input and did not respond to noxious chemical or mechanical stimuli applied to the nasal cavity, and (ii) wide dynamic range (WDR) neurones which responded to both noxious and nonnoxious chemical or mechanical stimuli applied to the nasal cavity.

Afferent Pathways↗

Facilitation of copulatory behavior by pCPA treatments following stria terminalis transection but not medial amygdala lesion in the male rat.

In this study, we examined the effect of p-chlorophenylalanine (pCPA), a serotonin synthesis inhibitor, on masculine sexual behavior in male rats following bilateral medial amygdala lesions (MAL) or bilateral stria terminals cuts (STC). Males with MAL and STC both showed a suppression of sexual behavior, compared to that of sham-operated males, when injected with saline. On the other hand, intraperitoneal injections of pCPA increased mount and intromission frequencies in males with STC. Although pCPA injections also slightly increased mount frequency in males with MAL, the injections failed to affect intromission behavior in MAL males. These suggest that the stria terminalis is insufficient as pathway of a whole output of the medial amygdala in regulating copulatory behavior. The medial amygdala may be involved in some other function, such as erection, than that of the stria terminalis.

Amygdala↗

An ephemeral pheromone of female house mice: perception via the main and accessory olfactory systems.

Two experiments examined the chemosensory modalities by which males detect an ephemeral sex pheromone in the freshly voided urine of female mice. Experiment 1 examined the interaction of deafferenting the accessory olfactory system (vomeronasal organ removal) and subsequent sexual experience upon ultrasonic vocalizations by male mice to freshly voided female urine. In general, sexually experienced males vocalized substantially more than sexually naive males. In addition, males possessing a vomeronasal organ vocalized slightly more than those without. Nonetheless, a functioning vomeronasal organ clearly was not essential for vocalizing to fresh female urine. Experiment 2 examined the effects of deafferenting the main olfactory system (ZnSO4 nasal irrigation) and/or the accessory olfactory system (vomeronasal removal) in sexually experienced males. Males with both olfactory systems functioning vocalized at high levels to fresh urine, while males with only one functioning system vocalized at intermediate levels. Males with neither system functioning did not vocalize at all to fresh urine. In contrast, when female mice themselves served as stimuli, all groups of males vocalized at high levels. We conclude that adult male mice can detect the ephemeral pheromone via either the main olfactory system or the accessory olfactory system. However, vocalizations to the female herself can be mediated by other sensory systems as well.

Animals↗

The dorsal diencephalic conduction system: a review of the anatomy and functions of the habenular complex.

The first part of this paper is an attempt to sketch an outline of the anatomy of the dorsal diencephalic conduction system by reviewing experimental evidence establishing the afferent and efferent connections of the habenular complex. This system provides an alternative to the descending medial forebrain bundle for the conduction of information from the limbic forebrain to limbic midbrain areas. The second part is a critical examination of experiments using ablation or electrical and chemical stimulation techniques which are concerned with the behavioural functions of the habenular complex. The habenula has been shown to play an important role in a diverse set of behavioural systems, which include olfaction, ingestion, mating, endocrine function, aversive motivation, and brain stimulation. Anatomical and behavioural support is presented for the view that the dorsal diencephalic conduction system provides an opportunity for interaction of activity in motivational systems with movement systems in the striatum and midbrain.

Animals↗

Developmental plasticity of the rat olfactory receptor sheet as shown by complete recovery of surface area and cell number from extensive early hypothyroid growth retardation.

To assess the effects of early thyroid deficiency, and recovery from this condition on growth and development of olfactory epithelium (OE), male Sprague-Dawley rat pups were rendered hypothyroid by addition of propylthiouracil (PTU) to their drinking water from birth. At weaning some rats continued to receive PTU while others ere allowed to recover by withdrawal of PTU. Body weights and plasma thyroxine levels were determined in all groups. At the ages of 25, 50 and 90 days, the OE of these hypothyroid and 'recovery' rats were compared with age-matched controls for surface area, epithelial thickness, density and total number of olfactory receptor neurons, basal cells and supporting cells, using morphometric and cell counting methods. Normal rats showed marked and highly significant increases in the OE surface area and olfactory neuron number (2.6- and 2.3-folds) during the post-weaning period. In the hypothyroid rats, body growth and thyroxine levels were severely suppressed. The OE in the 25-day-old hypothyroid rats showed more than 40% reduction in surface area and cell number, compared to controls, but mean epithelial thickness and surface density of cells were unchanged. In the post-weaning hypothyroid rats, the expansion of surface area was severely retarded, and increase in cell number ceased entirely. In rats allowed to recover by PTU withdrawal, by 90 days of age, body weight and size had markedly increased but had not caught up completely; however, thyroxine levels were restored to normal and the surface area and cell number in the OE had increased in a compensatory manner, completely restoring the deficiencies in OE growth, including surface area, numbers of receptor neurons, basal cells and supporting cells. The results indicate marked growth plasticity of OE in the post-weaning rats. This pronounced ability to recover from early growth retardation contrasts with that seen in central neural structures, and indicates the great potential of OE for use as a model neural system for the study of recovery from early damage and growth retardation.

Animals↗

The principal pathway from the piriform cortex to the deep amygdaloid nuclei in the cat.

Pathways relaying olfactory information to the deep amygdaloid nuclei (AMYGd) were analyzed with electrophysiological techniques in anesthetized cats. Stimulation of the piriform cortex (PC) produced orthodromic spikes in some AMYGd neurons with a mean latency of 12.2 ms and antidromic responses in other neurons with a mean latency of 9.3 ms. Stimulation of the AMYGd produced antidromic spikes in some PC neurons with a mean latency of 11.5 ms. Some neurons in the entorhinal area (EA) were activated orthodromically from the PC with a mean latency of 22 ms, and a proportion of these cells was also activated antidromically from the AMYGd. Some neurons in the agranular insular cortex were activated orthodromically from the PC, but none of them responded antidromically to AMYGd stimulation. From these observations, it is suggested that olfactory information reaches the AMYGd directly from the PC or indirectly via the EA, and that the direct path conveys the major olfactory input from the PC to the AMYGd.

Amygdala↗

An integrative hypothesis concerning the pathogenesis and progression of Alzheimer's disease.

Observations, in Alzheimer's disease, in the pattern of nerve cell damage and loss, the pathology, microchemistry and immunology of senile plaques and neurofibrillary tangles and alterations in blood vessels are drawn together into a hypothesis that attempts to explain the pathogenesis and progression of the disorder. At the heart of this hypothesis lies a defect in blood brain barrier function and/or structure within the cerebral cortex and this defect may be the cause of the cerebral vessel amyloidosis common in many patients with Alzheimer's disease. Age-related alterations in blood brain barrier allow for damage to nerve terminals and limited formation of senile plaques within cerebral cortex; neurofibrillary tangles are formed within cortical and subcortical nerve cells which project to or near damaged vessels/senile plaques. Uptake of "neurotoxin" at affected terminals and retrograde transport to perikarya causes neurofibrillary tangles to be formed; their accumulation leads to perikaryal changes culminating in cell death and loss. Loss of cells in cortically projecting areas of subcortex such as nucleus basalis, locus caeruleus and dorsal raphe, which terminate on cerebral vessels, causes further blood brain barrier dysfunction, new plaque formation and continued cell loss in cortex and subcortex. Once started, such a process could be self-perpetuating and the initial site of damage could lie within the amygdala/hippocampus with putative pathogenic agent accessing the brain via the olfactory pathways.

Age Factors↗

Persistent changes in behaviour and brain serotonin during ageing in rats subjected to infant nasal virus infection.

Suckling rats were infected intranasally with the temperature-sensitive mutant G41 strain of vesicular stomatitis virus. The rats survived but demonstrated lifelong learning deficits in the Morris maze and impaired exploratory behaviour in the open field test. When examined at 18 months of age they had a severe loss of neurons in the medial and dorsal raphe nuclei in the brain stem and reduced levels of serotonin and its metabolite 5-hydroxyindole acetic acid in the cerebral neocortex and hippocampus. The levels of noradrenaline, dopamine, homovanillic acid, 3,4-dihydroxyphenylacetic acid, choline acetyltransferase and glutamate decarboxylase were largely unaffected. The permanent disturbance in brain serotonin metabolism did not cause any histological changes in the cerebral cortex. Thus there were no neurofibrillary tangles or amyloid plaques as has been reported as a late effect of chemically induced lesion to the cholinergic system in the rat brain. It is concluded that the brain serotonergic system is especially vulnerable to an episode of virus attack along olfactory pathways and that the neurochemical and behavioural alterations caused by such an episode persist during a major part of the animal's life span.

Aging↗

Interamygdaloid connections in the rat studied by the horseradish peroxidase method.

Neurons of the rat amygdaloid body were labeled with horseradish peroxidase following its injection into contralateral nuclei of the amygdala. The results strongly suggest that there is a contralateral amygdaloid projection from the basal (dorsal and ventral) nuclei of amygdala; it terminates in the medial, central and lateral nucleus. True commissural connections were found only between posterior parts of the cortical nuclei of amygdala and between homonymous areas of the piriform cortex.

Amygdala↗

The comparative distribution of forebrain receptors for neurohypophyseal peptides in monogamous and polygamous mice.

Several recent studies have suggested that the neurohypophyseal peptide oxytocin may have a role within the brain to mediate various forms of affiliative behavior. As the regulation of oxytocin function may be largely determined by the number and distribution of its membrane bound receptor, we investigated oxytocin receptor distribution in two Peromyscus species selected for differences in affiliative behavior. Using in vitro receptor autoradiography with the selective oxytocin receptor ligand [125I]d(CH2)5[Tyr(Me)2,Tyr-NH9(2)]OVT ([125I]OTA), we compared Peromyscus maniculatus, a polygamous species, to Peromyscus californicus, a monogamous species. Marked species differences in the distribution of [125I]OTA were apparent in several brain areas, including olfactory pathways, bed nucleus of the stria terminalis, amygdala, dorsal lateral septum, and several cortical regions. In addition, gender differences in the binding pattern were evident in several regions, mostly due to sexually dimorphic patterns in the polygamous species, P. maniculatus. To further compare these species, the binding of a [3H]arginine-vasopressin antagonist was assessed in alternate sections from those used for [125I]OTA. Relative to oxytocin receptors, binding to arginine-vasopressin receptors showed fewer species differences, although the monogamous species appeared to have more arginine-vasopressin receptors in the neocortex and lateral septum. The striking differences in oxytocin receptor distribution are consistent with earlier studies in other rodents, suggesting that oxytocin may have an important role for mediating species-typical patterns of social affiliation.

Animals↗

Immunohistochemical and behaviour pharmacological analysis of rats inoculated intranasally with vesicular stomatitis virus.

A temperature-sensitive mutant of vesicular stomatitis virus was inoculated intranasally into infant Sprague-Dawley rats aged 9 to 17 days. Rats receiving the virus at 9 days of age had an extensive spread of infection throughout the brain and the animals died after a few days. Rats inoculated at day 11 postnatally survived and the infection was limited to the olfactory pathways, hypothalamus, diagonal bands and the anterior raphe nuclei. Stereological measurements showed that the volume of infected neurons constituted 67 +/- 10% of the total neuronal volume in the dorsal raphe nucleus. Double-labelling experiments revealed that both 5-hydroxytryptamine- and substance P-immunoreactive neurons contained the virus antigen. The motor stimulant effect of amphetamine was studied at 3 months post infection. The increase in amphetamine-induced frequency and duration of rearing was significantly attenuated in infected rats and the amphetamine-induced locomotion was slightly reduced.

Administration, Intranasal↗

Implantation of a scaffold following bulbectomy induces laminar organization of regenerating olfactory axons.

Primary olfactory axons expressing different odorant receptors are interspersed within the olfactory nerve. However, upon reaching the outer nerve fiber layer of the olfactory bulb they defasciculate, sort out, and refasciculate prior to targeting glomeruli in fixed topographic positions. While odorant receptors are crucial for the final targeting of axons to glomeruli, it is unclear what directs the formation of the nerve fiber and glomerular layers of the olfactory bulb. While the olfactory bulb itself may provide instructive cues for the development of these layers, it is also possible that the incoming axons may simply require the presence of a physical scaffold to establish the outer laminar cytoarchitecture. In order to begin to understand the underlying role of the olfactory bulb in development of the outer layers of the olfactory bulb, we physically ablated the olfactory bulbs in OMP-IRES-LacZ and P2-IRES-tau-LacZ neonatal mice and replaced them with artificial biological scaffolds molded into the shape of an olfactory bulb. Regenerating axons projected around the edge of the cranial cavity at the periphery of the artificial scaffold and were able to form an olfactory nerve fiber layer and, to some extent, a glomerular layer. Our results reveal that olfactory axons are able to form rudimentary cytoarchitectonic layers if they are provided with an appropriately shaped biological scaffold. Thus, the olfactory bulb does not appear to provide any tropic substance that either attracts regenerating olfactory axons into the cranial cavity or induces these axons to form a plexus around its outer surface.

Animals↗

Behavioral analysis of olfactory coding and computation in rodents.

Behavioral analysis is essential to understand how the olfactory system transforms chemosensory signals into information that can be used to guide actions. Recent studies in rodents have begun to address the behavioral relevance of putative olfactory codes and computations including spatial maps, oscillatory synchrony, and evolving temporal codes. To date, these studies have failed to find support for a role of any of these mechanisms in odor discrimination. Progress calls for experiments using precise psychophysical methods in conjunction with neural recording or perturbation, in addition to ethologically minded exploration of more complex forms of odor-guided behavior.

Action Potentials↗

Neuronal development: specifying a hard-wired circuit.

The formation of neuronal circuits that relay distinct olfactory information is thought to depend on cues provided by pre-synaptic receptor neurons. But direct visualization of second order neurons in Drosophila now suggests that dendritic targeting occurs independently of interactions with incoming sensory neurons.

Animals↗

Molecular, anatomical, and functional organization of the Drosophila olfactory system.

BACKGROUND: Olfactory receptor neurons (ORNs) convey chemical information into the brain, producing internal representations of odors detected in the periphery. A comprehensive understanding of the molecular and neural mechanisms of odor detection and processing requires complete maps of odorant receptor (Or) expression and ORN connectivity, preferably at single-cell resolution. RESULTS: We have constructed near-complete maps of Or expression and ORN targeting in the Drosophila olfactory system. These maps confirm the general validity of the "one neuron--one receptor" and "one glomerulus--one receptor" principles and reveal several additional features of olfactory organization. ORNs in distinct sensilla types project to distinct regions of the antennal lobe, but neighbor relations are not preserved. ORNs grouped in the same sensilla do not express similar receptors, but similar receptors tend to map to closely appositioned glomeruli in the antennal lobe. This organization may serve to ensure that odor representations are dispersed in the periphery but clustered centrally. Integrated with electrophysiological data, these maps also predict glomerular representations of specific odorants. Representations of aliphatic and aromatic compounds are spatially segregated, with those of aliphatic compounds arranged topographically according to carbon chain length. CONCLUSIONS: These Or expression and ORN connectivity maps provide further insight into the molecular, anatomical, and functional organization of the Drosophila olfactory system. Our maps also provide an essential resource for investigating how internal odor representations are generated and how they are further processed and transmitted to higher brain centers.

Animals↗

Genetic and functional subdivision of the Drosophila antennal lobe.

Olfactory systems confer the recognition and discrimination of a large number of structurally distinct odor molecules. Recent molecular analysis of odorant receptor (OR) genes and circuits has led to a model of odor coding in which a population of olfactory sensory neurons (OSNs) expressing a single OR converges upon a unique olfactory glomerulus. Activation of the OR can thus be read out by the activation of its cognate glomerulus. Drosophila is a powerful system in which to test this model because the entire repertoire of 62 ORs can be manipulated genetically. However, a complete understanding of how fly olfactory circuits are organized is lacking. Here, we present a nearly complete map of OR projections from OSNs to the antennal lobe (AL) in the fly brain. Four populations of OSNs coexpress two ORs along with Or83b, and a fifth expresses one OR and one gustatory receptor (GR) along with Or83b. One glomerulus receives coconvergent input from two separate populations of OSNs. Three ORs label sexually dimorphic glomeruli implicated in sexual courtship and are thus candidate Drosophila pheromone receptors. This olfactory sensory map provides an experimental framework for relating ORs to glomeruli and ultimately behavior.

Animals↗