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Localization of cannabinoid receptor mRNA in rat brain.

Cannabinoid receptor mRNA was localized in adult rat brain by 35S-tailed oligonucleotide probes and in situ hybridization histochemistry. Labelling is described as uniform or non-uniform depending on the relative intensities of individual cells expressing cannabinoid receptor mRNA within a given region or nucleus. Uniform labelling was found in the hypothalamus, thalamus, basal ganglia, cerebellum and brainstem. Non-uniform labelling that resulted from the presence of cells displaying two easily distinguishable intensities of hybridization signals was observed in several regions and nuclei in the forebrain (cerebral cortex, hippocampus, amygdala, certain olfactory structures). Olfactory-associated structures, basal ganglia, hippocampus, and cerebellar cortex displayed the heaviest amounts of labelling. Many regions that displayed cannabinoid receptor mRNA could reasonably be identified as sources for cannabinoid receptors on the basis of well documented hodologic data. Other sites that were also clearly labelled could not be assigned as logical sources of cannabinoid receptors. The localization of cannabinoid receptor mRNA indicates that sensory, motor, cognitive, limbic, and autonomic systems should all be influenced by the activation of this receptor by either exogenous cannabimimetics, including marijuana, or the yet unknown endogenous "cannabinoid" ligand.

Animals↗

The development of olfactory and hippocampal pathways in the brain of the rat.

The temporal order of development of olfactory, hippocampal and thalamocortical connections has been determined by light microscopy. Scalpel lesions were made to interrupt these connections and the resulting terminal degeneration was stained by Eager's method (1970). A post-operative survival time of one to four days was used. Evidence of the development of these connections was first obtained at the following ages: Olfactory mucosa to olfactory bulb: axon fascicles by 16 days of gestation and terminals in glomeruli at birth; Olfactory bulb to prepyriform cortex at birth; Prepyriform to entorhinal cortex at 13 days after birth; Entorhinal cortex to hippocampus (the perforant path) at 9 days; Hippocampal dentate-Ammonic mossy fibres at 9 days; Hippocampal efferent projection to the septum at birth; Subicular projections to the anterior thalamus at birth and to the mammillary body at 6 days; Hippocampal commissural connections at birth; Corticothalamic and thalamocortical connections by 2 days. These results are discussed in relation to the question of how the development of brain connections is programmed.

Animals↗

Computational parallels between the biological olfactory pathway and its analogue 'the electronic nose': Part II. Sensor-based machine olfaction.

Over the last fifteen years, we have witnessed a rapid expansion in the development of artificial odour sensing systems, or so called 'electronic nose' systems. Whilst the power of this approach to flavour has undoubtedly been demonstrated by its recent application to various complex odours, it will be argued that the original research programme, aimed at developing a comparative model of the biological olfactory pathway, has degenerated into an attempt to obtain an ad hoc workable system, based around readily available sensor and pattern recognition (PARC) technologies. At the time, the first 'model' nose system reflected the limited understanding of sensory information processing carried out within the biological olfactory pathway. We are now presented with an opportunity to evaluate and re-assess the architecture for an electronic nose, in view of the recent advances in understanding the key processing principals exploited by the olfactory bulb and cortex in the identification and characterisation of molecular stimuli. In Part II of this paper, we examine the parallels that exist between the biological olfactory system and the electronic nose. It is shown that the two systems share many similarities in their architectures and other properties, such as odour delivery, nonspecific sensor/receptor response, sensor/receptor preprocessing and content addressable memory (CAM) function. Of particular importance, both systems need to overcome similar operating problems, such as sensor/receptor drift, degeneration and poisoning, limited sensor/receptor sensitivity, discrimination of odour quality invariant of intensity and also the identification of particular odour components within a mixture of background odours. Finally, a number of opportunities for improving the biological plausibility of electronic nose systems are suggested that may yield an improvement in performance.

Animals↗

Characterization of NMDA receptor subunit-specific antibodies: distribution of NR2A and NR2B receptor subunits in rat brain and ontogenic profile in the cerebellum.

Selective antisera for NMDA receptor subunits NR2A and NR2B have been developed. Each antiserum identifies a single band on an immunoblot at approximately 175 kDa that appears to be the appropriate subunit of the NMDA receptor. Using these antisera the relative densities of the subunits in eight areas of adult rat brain have been determined. The NR2A subunit was found to be at its highest level in hippocampus and cerebral cortex, to be at intermediate levels in striatum, olfactory tubercle, mid-brain, olfactory bulb, and cerebellum, and to be at lowest levels in the pons-medulla. The NR2B subunit was found to be expressed at its highest levels in the olfactory tubercle, hippocampus, olfactory bulb, and cerebral cortex. Intermediate levels were expressed in striatum and mid-brain, and low levels were detected in the pons-medulla. No signal for NR2B was found in the cerebellum. These regional distributions were compared with that for [3H]MK-801 binding sites. It was found that although the distribution of the NR2A subunit corresponds well with radioligand binding, the distribution of the NR2B subunit does not. The ontogenic profiles of NR2A and NR2B subunits in the rat cerebellum were also determined. Just following birth [postnatal day (P) 2] NR2A subunits are undetectable, whereas NR2B subunits are expressed at amounts easily measurable. Beginning at about P12 the levels of NR2A rise rapidly to reach adult levels by P22. At the same time (P12), levels of NR2B protein begin to decline rapidly to reach undetectable levels by 22 days after birth.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

The role of the olfactory tubercle in the effects of cocaine, morphine and brain-stimulation reward.

Using the quantitative 2-[14C]deoxyglucose autoradiographic method, local rates of glucose utilization were measured in rats after the administration of morphine or cocaine in the presence or absence of rewarding brain stimulation to the medial forebrain bundle. In animals that did not receive brain stimulation, cocaine significantly increased glucose utilization in the olfactory tubercle, medial prefrontal cortex and substantia nigra pars reticulata, whereas morphine significantly increased glucose metabolism in the olfactory tubercle only. Stimulation itself increased metabolic rates in a number of sites, such as the olfactory tubercle, nucleus accumbens, medial prefrontal cortex, ventral tegmental area and others. However, in self-stimulating animals both morphine and cocaine caused further increases in activity in the olfactory tubercle. Since the olfactory tubercle was the only structure to cause a significant increase in metabolic rate following each treatment, the results implicate this limbic structure in the rewarding effects of morphine, cocaine and brain-stimulation reward.

Animals↗

Eye closure in darkness animates olfactory and gustatory cortical areas.

In two previous fMRI studies, it was reported that eyes-open and eyes-closed conditions in darkness had differential effects on brain activity, and typical patterns of cortical activity were identified. Without external stimulation, ocular motor and attentional systems were activated when the eyes were open. On the contrary, the visual, somatosensory, vestibular, and auditory systems were activated when the eyes were closed. In this study, we investigated whether cortical areas related to the olfactory and gustatory system are also animated by eye closure without any other external stimulation. In a first fMRI experiment (n = 22), we identified cortical areas including the piriform cortex activated by olfactory stimulation. In a second experiment (n = 12) subjects lying in darkness in the MRI scanner alternately opened and closed their eyes. In accordance to previous studies, we found activation clusters bilaterally in visual, somatosensory, vestibular and auditory cortical areas for the contrast eyes-closed vs. eyes-open. In addition, we were able to show that cortical areas related to the olfactory and gustatory system were also animated by eye closure. These results support the hypothesis that there are two different states of mental activity: with the eyes closed, an "interoceptive" state characterized by imagination and multisensory activity and with the eyes open, an "exteroceptive" state characterized by attention and ocular motor activity. Our study also suggests that the chosen baseline condition may have a considerable impact on activation patterns and on the interpretation of brain activation studies. This needs to be considered for studies of the olfactory and gustatory system.

Attention↗

Olfactory bulb projections to the parahippocampal area of the rat.

Recent evidence suggests that the main olfactory bulb projects caudally beyond the prepiriform cortex and the cortical amygdaloid nuclei to the region of the piriform lobe called the parahippocampal area. Included within this area is the entorhinal cortex, which is composed of six major subdivisions. Since questions remain as to which of these subdivisions receives centripetal fibers from the bulb, we reexamined these projections using autoradiography and HRP histochemistry and correlated the sites of termination with the cytoarchitecture of the entorhinal cortex. The results indicate that olfactory bulb axons reach all parts of the parahippocampal area, including the cortex which forms the medial and lateral banks of the amygdaloid sulcus (area TR), and both subdivisions of the laterally located entorhinal cortex (28L' and 28L). Also, label is observed over the more medially located fields of the entorhinal cortex, including the cortex posterior to the cortical amygdaloid nucleus (28M'), as well as the ventrolateral parts of medial entorhinal cortex (28M). In addition, evidence of label occurs over the full extent of the transition zone (28i) which separates areas 28L and 28M. These results suggest that the olfactory bulb has a more extensive projection to the parahippocampal area in the rat than previously thought, and may provide at least some input to all of the parahippocampal areas which project to the hippocampal formation.

Animals↗

The differential projections of the olfactory bulb and accessory olfactory bulb in mammals.

Three species were studied, the rabbit, opossum and rat. Lesions of the main olfactory bulb caused terminal degeneration, assayed by the Fink-Heimer method, to occur in the ipsilateral olfactory tubercle, prepyriform cortex (including its periamygdaloid part), ventrolateral entorhinal area, and in anterior and posterolateral divisions of the cortical amygdaloid nucleus. The various parts of the ipsilateral anterior olfactory nucleus and the rostroventral end of the anterior continuation of the hippocampus (hippocampal rudiment) also received this projection. Lesions of the accessory olfactory bulb, which receives its sensory input from the vomeronasal (Jacobson's) organ, caused terminal degeneration to occur in the medial amygdaloid nucleus and in a posteromedial part of the cortical amygdaloid nucleus. This projection was conveyed by an accessory olfactory tract, which is accompanied in part of its course by a small nucleus, the bed nucleus of the accessory olfactory tract. The accessory olfactory tract is initially a part of the lateral olfactory tract but becomes increasingly indivuated at more posterior levels. It parts company with the lateral olfactory tract at the rostral end of the amygdaloid region, and, in addition to distributing to the medio-cortical amygdaloid region, it enters the stria terminalis to terminate in the bed nucleus of the stria terminalis in a small region bearing cytoarchitectonic resemblance to the medial amygdaloid nucleus. The topographic segregation of the areas of termination of the olfactory and accessory olfactory (vomeronasal) projections is suggestive of a functional dichotomy in the organization of the olfactory system...

Amygdala↗

Expression of the gene encoding the chemorepellent semaphorin III is induced in the fibroblast component of neural scar tissue formed following injuries of adult but not neonatal CNS.

This study evaluates the expression of the chemorepellent semaphorin III (D)/collapsin-1 (sema III) following lesions to the rat CNS. Scar tissue, formed after penetrating injuries to the lateral olfactory tract (LOT), cortex, perforant pathway, and spinal cord, contained numerous spindle-shaped cells expressing high levels of sema III mRNA. The properties of these cells were investigated in detail in the lesioned LOT. Most sema III mRNA-positive cells were located in the core of the scar and expressed proteins characteristic for fibroblast-like cells. Neuropilin-1, a sema III receptor, was expressed in injured neurons with projections to the lesion site, in a subpopulation of scar-associated cells and in blood vessels around the scar. In contrast to lesions made in the mature CNS, LOT transection in neonates did not induce sema III mRNA expression within cells in the lesion and was followed by vigorous axonal regeneration. The concomitant expression of sema III and its receptor neuropilin-1 in the scar suggests that sema III/neuropilin-1-mediated mechanisms are involved in CNS scar formation. The expression of the secreted chemorepellent sema III following CNS injury provides the first evidence that chemorepulsive semaphorins may contribute to the inhibitory effects exerted by scars on the outgrowth of injured CNS neurites. The vigorous regrowth of injured axons in the absence of sema III following early neonatal lesions is consistent with this notion. The inactivation of sema III in scar tissue by either antibody perturbation or by genetic or pharmacological intervention could be a powerful means to promote long-distance regeneration in the adult CNS.

Animals↗

Autoradiographic visualization of a calcium channel antagonist, [125I]omega-conotoxin GVIA, binding site in the brains of normal and cerebellar mutant mice (pcd and weaver).

An in vitro autoradiographic technique has been used to localize [125I]omega-conotoxin GVIA binding sites in the brains of normal and cerebellar mutant mice. In the brains of normal mice, the highest densities of binding sites were observed at glomeruli of the olfactory bulb, cerebral cortex, caudate nucleus-putamen, hippocampus, and the nucleus of the solitary tract. Moderate densities of the silver grains occurred on the granular layer of the olfactory bulb, the molecular layer of the dentate gyrus, the molecular layer of the cerebellum, and the cochlear nucleus. No specific binding appeared in the white matter or the deep nucleus of the cerebellum, the corpus callosum, the internal capsule and the external plexiform layer of the olfactory bulb. Autoradiographic studies of the cerebella of Purkinje cell degeneration (pcd) mice showed that the distribution of binding sites on the molecular layer of the cerebellum are not affected by the degeneration of Purkinje cells. However, only background levels of the silver grains occurred on the cerebella of agranular weaver mutant mice, suggesting that the receptors for omega-conotoxin GVIA in the cerebellum are predominantly distributed on the parallel fibers of granule cells.

Animals↗

Localization of trkB mRNA in postnatal brain development.

We investigated the localization of trkB mRNA, which encodes a putative component of high-affinity brain-derived neurotrophic factor (BDNF) or the neurotrophin-3 (NT-3) receptor, in the postnatal rat brain by in situ hybridization histochemistry. At birth, trkB mRNA was strongly expressed in various regions with the thalamus and cerebral cortex showing the strongest expression. As the rat grows, expression generally persisted or declined in most regions with the exception of the hippocampus where trkB mRNA expression increased during postnatal development. In the adult brain, trkB mRNA was detected in the olfactory system, cerebral cortex, hippocampal formation, amygdala, and cerebellar cortex. These findings, together with the developmental profiles of BDNF and NT-3 mRNA expressions, suggest that trkB product (gp145trkB) mainly transduces NT-3 signals early in the postnatal period, and BDNF signals later in the period.

Aging↗

Three-dimensional metabolic mapping of the freeze-trapped brain: effects of ischemia in the Mongolian gerbil.

The effects of unilateral carotid artery (left or right) occlusion on the three-dimensional metabolic mapping were studied in the Mongolian gerbil (Meriones unguiculatus). The brain was freeze-trapped by the liquid nitrogen funnel technique and was analyzed for the two-dimensional distribution of the reduced pyridine nucleotides and oxidized flavoprotein using a time-sharing micro-light guide surface fluorometer/reflectometer. The results could be summarized as follows. Bilateral carotid artery occlusion induced uniform effects in terms of redox state in the cerebral cortex and the olfactory bulbs. After unilateral carotid occlusion, the redox state of the ischemic ipsilateral hemisphere was higher and was not affected by blood volume changes as evaluated from the reflectance signal scanned in several brains. The narrow band of tissue near the midline of the ischemic hemisphere connected to the contralateral hemisphere blood supply appeared in a large number of gerbils. The anomaly in the redox state of the intracellular space also appeared in subcortical structures, as seen in all the two-dimensional mappings measured. It seems that blood circulation to the two olfactory bulbs is connected to the same blood vessel. It is suggested that unilateral carotid artery-occluded gerbils may show large variability inside the same hemisphere and therefore results must be evaluated very carefully.

Animals↗

Correlations between unit firing and EEG in the rat olfactory system.

The olfactory EEG of awake animals displays oscillatory bursts of activity in the gamma- (30-100 Hz) range. The bursts are correlated with inflow of air over the receptor layer in the nose. None of the inputs to the cortices that display these oscillations carries periodic signals in the gamma-range. Thus these bursts are generated locally, either by neuronal feedback interactions or by coupling of oscillatory neurons. In the first case if the oscillations are generated by negative feedback, then two classes of cells must exist: excitatory neurons and inhibitory neurons with the same frequency of oscillation but with a quarter cycle phase lag by the inhibitory cells from the excitatory cells. On the other hand, if the EEG's result from coupling of cells that are intrinsically oscillatory, there should be a broad but monomodal distribution of phase values. In order to determine the origin of these bursts, we performed simultaneous recordings of EEG and multi-unit spikes in the 4 parts of the olfactory system (olfactory bulb, anterior olfactory nucleus, prepyriform cortex and lateral entorhinal area) of awake and motivated rats. For each sample, the EEG and the multi-unit spikes were recorded from the same local neighborhood. The multi-unit electrode recorded pulses from the principal output neurons of the respective cortical areas. In all locations tested, the oscillations in pulse probabilities of firing were found to have the same frequency as the dominant EEG frequency. In all 4 structures two sets of cells were found. One set displayed pulses in phase with the EEG and the other set displayed pulses that led or lagged the EEG by approximately 1/4 cycle. These data confirm the negative feedback interaction model rather than the coupled oscillator model for the generation of the bursts in the olfactory system. The relevance of these findings to other cortical systems, in casu the visual cortex is discussed.

Animals↗

Differential distribution of four hyperpolarization-activated cation channels in mouse brain.

Hyperpolarization-activated cation currents, termed I(h), are observed in a variety of neurons. Four members of a gene family encoding hyperpolarization-activated cyclic-nucleotide-gated cation channels (HCN1-4) have been cloned. The regional expression and cellular localization of the four HCN channel types in mouse brain was investigated using in situ hybridization. The expression of HCN1 was restricted to the olfactory bulb, cerebral cortex, hippocampus, superior colliculus and cerebellum. In contrast, HCN2 transcripts were found at high levels nearly ubiquitously in the brain, and the strongest signals were seen in the olfactory bulb, hippocampus, thalamus and brain stem. HCN3 was uniformly expressed at very low levels throughout the brain. Finally, HCN4 transcripts were prominently expressed selectively in the thalamus and olfactory bulb. Some neurons expressed two or more HCN channel transcripts including hippocampal pyramidal neurons (HCN1, HCN2 and low levels of HCN 4) and thalamic relay neurons (HCN2 and HCN4). Our results demonstrate that each HCN channel transcript has a unique distribution in the brain. Furthermore, they suggest that the heterogeneity of neuronal I(h) may be, at least in part, due to the differential expression of HCN channel genes.

Amino Acid Sequence↗

Expression of the Per1 gene in the hamster: brain atlas and circadian characteristics in the suprachiasmatic nucleus.

Recent progress in study on the molecular component of mammalian clocks has claimed that mammals and Drosophila share the similar fundamental clock oscillating system. In the present study, we investigated expression of Per1, the first gene of the mammalian homolog of the Drosophila clock gene period, in the hamster brain, and we also examined its circadian expression pattern in the mammalian clock center, the suprachiasmatic nucleus (SCN). In situ hybridization using isotope-labeled cRNA probes revealed a wide and region-specific distribution of Per1 in the hamster brain and spinal cord. High levels of Per1 were found in the internal granular layer of the granular cells of the olfactory bulb, anterior olfactory nuclei, tenia tecta, olfactory tubercle, piriform cortex, suprachiasmatic nucleus, and gyrus dentatus of hippocampus. Moderate levels of expression were detected in many brain regions including the granular layer of the cerebellum, anterior paraventricular thalamic nucleus, caudate-putamen, inferior colliculus, pontine nuclei, inferior olive, and nucleus of the solitary tract. We examined the circadian profile of hamster Per1 mRNA in the SCN in constant darkness and found that Per1 expression showed a peak at subjective day (circadian time [CT] 4) and formed a trough at subjective night (CT16-CT20). A brief exposure of light at CT16 could acutely induce large quantities of Per1 mRNA in the hamster SCN, except for its dorsomedial subdivision. These findings suggest that the characteristics of Per1 gene expression in the mammalian circadian center (showing a peak in the daytime and a trough in the nighttime and a rapid inducibility by light) are common among mammalian species. Lastly, in hamster brain, Per1 gene is also inducible in extra-SCN brain nuclei, since light at night also elicited Per1 mRNA in neurons of the hypothalamic paraventricular nucleus.

Animals↗

Computational parallels between the biological olfactory pathway and its analogue 'the electronic nose': Part I. Biological olfaction.

Over the last fifteen years, we have witnessed a rapid expansion in the development of artificial odour sensing systems, or so called 'electronic nose' systems. Whilst the power of this approach to flavour analysis has undoubtedly been demonstrated by its recent application to various complex odours, it will be argued that the original research programme, aimed at developing a comparative model of the biological olfactory pathway, has degenerated into an attempt to obtain an ad hoc workable system, based around readily available sensor and pattern recognition (PARC) technologies. At the time, the first 'model' nose system reflected the limited understanding of sensory information processing carried out within the biological olfactory pathway. We are now presented with an opportunity to evaluate and re-assess the architecture for an electronic nose, in view of the recent advances in understanding the key processing principals exploited by the olfactory bulb and cortex in the identification and characterisation of molecular stimuli. In Part I of this paper, the rapid developments in the understanding of the information processing performed by the biological olfactory system are critically reviewed, and its relevance to current research in artificial olfaction is considered. Not only have the initial biochemical pathways involved in the transduction of odour stimuli been uncovered, but also computational models of the key synaptic circuits have advanced to the point where network simulations are clearly capable of odour discrimination. The key processing principles exploited in the olfactory pathway for overcoming operating constraints such as sensor drift/degeneration, limited sensitivity, and xenobiotic response are highlighted, so that their integration into the electronic analogue may be explored in Part II.

Computer Simulation↗

Changes in c-fos mRNA expression in rat brain during odor discrimination learning: differential involvement of hippocampal subfields CA1 and CA3.

Levels of c-fos mRNA were measured with in situ hybridization to test for behaviorally dependent changes in neuronal activity in three subdivisions of hippocampus and in components of the olfactory and visual systems. In rats that performed a well-learned nose-poke response for water reward, c-fos mRNA levels were broadly increased, relative to values in home cage-control rats, in visual cortex, superior colliculus, olfactory bulb, and, to comparable levels, regions CA3 and CA1 of hippocampus; hybridization was not increased in the dentate gyrus. In rats first trained on the nose-poke behavior and then required to discriminate between two odors for water reward, the increase in c-fos mRNA was generally not as great and was more regionally differentiated. Thus, in olfactory bulb, hybridization was more greatly elevated in lateral than medial fields, thereby exhibiting regional activation corresponding to the topographic representation of the predominant odor sampled in the discrimination task. In hippocampus of odor-discrimination rats, c-fos mRNA levels were far greater in the region CA3 than region CA1, but remained at cage control values in stratum granulosum. Interestingly, c-fos mRNA levels in each hippocampal subdivision were highly correlated with levels in other regions (e.g., visual cortex) for home cage controls but not for rats in the two behavioral groups. Thus, c-fos mRNA levels in cage-control rats appeared to be regulated by some generalized factor acting throughout much of the brain (e.g., arousal), while odor-discrimination performance changed the pattern of expression within hippocampus, and allowed for a differentiated response by olfactory regions to emerge. These findings suggest that hippocampus possesses multiple modes of functioning and makes contributions to behavior that vary according to task demands.

Animals↗