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The medial forebrain bundle of the rat. II. An autoradiographic study of the topography of the major descending and ascending components.

The medial forebrain bundle (MFB) is a complex fiber system that courses through and partly arises and partly terminates within the lateral preoptic and lateral hypothalamic areas. It consists mainly of thin fibers and may be comprised of as many as 50 descending and ascending components of varying lengths and of different origins and/or destinations (Nieuwenhuys et al., '82). With the aid of an an atlas of the MFB and the surrounding brain areas in the rat presented in the preceding paper (Nieuwenhuys et al., '82), the position and topographic relationships of some 21 components of the bundle have been analyzed in detail, in brains that had been prepared for autoradiography following injections of tritiated amino acids into a number of structures that are known to contribute fibers to the MFB. From this analysis it is clear that most of the labeled components occupy specific and rather constant positions within the MFB. For example, the ascending components are largely confined to the dorsal half of the bundle; those arising from the medial preoptic area and the various hypothalamic nuclei are distributed rather diffusely over much of the MFB; and the descending components that arise from the olfactory tubercle and the magnocellular preoptic nucleus are confined to restricted parts of the bundle. These findings indicate that the neurons which occupy different parts of the lateral hypothalamic area probably receive distinctive inputs, and to a first approximation these are likely to be determined principally by their position within the MFB.

Amygdala↗

Catecholaminergic innervation of the septal area in man: immunocytochemical study using TH and DBH antibodies.

The catecholaminergic innervation of the human septal area and closely related structures has been visualized by using tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH) as immunocytochemical markers. TH-like immunoreactivity with no corresponding DBH labelling was considered to be indicative of dopaminergic fibers. Catecholaminergic innervation offered the following similarities to that of rodents: moderate innervation in the medial septal division, with predominant DBH immunolabelling; dense dopaminergic innervation in the lateral septal nuclei, organized in a laminar pattern; presence of dopaminergic pericellular arrangements in the dorsal septum and bed n. of the stria terminalis; clustering of dopaminergic terminals in n. accumbens associated with a medioventral zone of DBH-like immunoreactive fibers; close overlap between dopaminergic fields and acetylcholinesterase-reactive zones in both the lateral septum and the n. of the stria terminalis. Differences with the catecholaminergic septal innervation of rodents consisted of general caudal extension of the dopaminergic fields, possibly accounted for by the vertical stretching and caudal displacement of the septal nuclei in man; complementary lateromedial topography of dopaminergic and DBH-immunoreactive inputs in the n. of the stria terminalis as opposed to their dorsoventral organization in rodents; presence of TH-immunolabelled cell group in the anterior olfactory nucleus and parolfactory cortex, which seems specific for primates. Precise topographical mapping of the catecholaminergic structures in this central region of the limbic forebrain seems to be a prerequisite for accurate tissue sampling in the biochemical investigations of pathological cases and should help in the interpretation of aminergic dysfunction in a variety of human diseases.

Acetylcholinesterase↗

Comparative aspects of the olfactory portion of the entorhinal cortex and its projection to the hippocampus in rodents, nonhuman primates, and the human brain.

The entorhinal cortex is a component of the hippocampal formation characterized by its projection to the dentate gyrus. Cortical input is relayed to the hippocampus through the entorhinal cortex. The entorhinal cortex can be recognized best in mammals, and it reaches its maximal importance in both human and nonhuman primates. In primates, but not in rodents, the olfactory projection from the olfactory bulb is the only direct sensory projection reaching the entorhinal cortex, where it defines a rostromedial subfield (olfactory subfield of the entorhinal cortex, EO). In this communication, we consider some comparative aspects of this projection in rodents, nonhuman primates, and human brains, in relation to the forward projection to the hippocampus. We base our analysis on the following three considerations: (1) Topology: Topologically, the entorhinal cortex lies medial to the rhinal fissure, well developed in all mammals except in humans. EO maintains the same topological relationship to the rhinal fissure in the three mammalian orders. (2) Cytoarchitecture: The rostromedial portion shows poorer lamination compared with caudomedial portions of the entorhinal cortex in the three mammalian orders examined. (3) Connectivity: In rodents, the olfactory projection covers the whole extent of the entorhinal cortex, whereas it decreases substantially in macaques, where it roughly covers 15% of the entorhinal cortex. The human EO as defined by topological and cytoarchitectonic criteria may be even less than in macaques. Interestingly, good evidence exists that all of these mammalian orders maintain the same topographical scheme of projection to the rostral end of the hippocampus.

Animals↗

The piriform cortex and the endopiriform nucleus in the rat reveal generally similar pattern of connections.

The afferent and efferent connections of the piriform cortex and the endopiriform nucleus in the rat were studied by the method of axonal transport of two fluorescent tracers: Fluoro-Gold and Fluoro-Ruby. The results indicate that both structures possess not only the connections with the olfactory system, but also the reciprocal connections with the limbic system (entorhinal cortex, amygdaloid body), thalamus (mediodorsal and midline nuclei), extrapyramidal system (ventral part of the nucleus accumbens). The topographic differences in the organization of the association connections between the anterior and posterior parts of the piriform cortex are reported. Additionally, the reciprocal and relatively numerous connections between the endopiriform nucleus and the piriform cortex may result in their modulatory function, which in some pathological circumstances may have a critical significance in epileptogenesis.

Afferent Pathways↗

Olfactory discrimination of amino acids in brown bullhead catfish.

Olfactory discrimination of amino acids was investigated in brown bullhead catfish (Ameiurus nebulosus). Based on the magnitude of the observed food search activity of catfish conditioned to single amino acids, the tested compounds were classified as being detected by the catfish as equal to, similar to, or different from the conditioned stimulus. L-Proline (L-Pro)-conditioned brown bullhead catfish discriminated all amino acids from L-Pro, but catfish conditioned to L-valine (L-Val) and L-isoleucine (L-Ile) did not discriminate L-Val from L-Ile nor L-Ile from L-Val; however, all other amino acids tested were always discriminated from these two compounds. Catfish conditioned to L-alanine (L-Ala) discriminated basic, acidic and several neutral amino acids with long side-chains (LCNs) from L-Ala; however, they did not always discriminate L-Ala from all neutral amino acids with short side-chains (SCNs). The L-norleucine (L-nLeu)-conditioned fish responded to L-norvaline (L-nVal), L-methionine (L-Met) and L-Ala similarly to L-nLeu, indicating that these amino acids are detected as similar or identical to L-nLeu. L-nLeu was, however, discriminated from L-Ala in L-Ala-conditioned catfish. Interestingly, L-leucine (L-Leu) was discriminated from the conditioned stimuli, L-Ala, L-Ile and L-Val, indicating independent receptors for L-Leu. Although conditioned catfish discriminated other amino acids from L-arginine hydrochloride (L-Arg), in some tests they were unable to discriminate L-Arg from L-lysine hydrochloride (L-Lys). These results imply the existence of independent olfactory receptive pathways for: (i) L-Pro; (ii) basic amino acids (L-Arg and L-Lys); (iii) L-Leu; (iv) other neutral amino acids with branched side-chains (L-Ile and L-Val); (v) neutral amino acids with long linear side-chains (L-nLeu, L-nVal and L-Met); (vi) neutral amino acids with short side-chains; and (vii) amino acids with sulfhydryl groups (L-Cys and L-homoCys).

Amino Acids↗

Cell migration from the transplanted olfactory placode in Xenopus.

The eye vesicle of Xenopus borealis has been replaced with the transplanted olfactory primordium from Xenopus laevis in an attempt to determine whether cells from the transplant could migrate along the regrowing olfactory nerve and become incorporated into the CNS of the host. The use of X. laevis and X. borealis pairs allowed us to distinguish the cells of the host from those of the donor at the cellular level by means of the characteristic fluorescent nuclear spots (Q bands) of X. borealis. Transplantation was performed on pairs of animals at stages 23/24. The olfactory anlage was readily incorporated into the host, often fusing with the host homolateral organ and inhibiting the regrowth of the eye vesicle. An olfactory nerve developed from the transplanted organ. In the majority of cases, the nerve reached the diencephalon at the level of entrance of the optic nerve. Along the nerve originating from the transplanted organ we observed a stream of cells with the characteristics of the donor. These cells penetrated the host's CNS and became incorporated into it. The nature of these cells has not been ascertained by specific neuronal markers. However, on the basis of their morphology and disposition, the hypothesis suggested is that some of the migrating cells are neurons.

Animals↗

Calretinin and calbindin-D28k in rat brain: patterns of partial co-localization.

Calretinin and calbindin-D28k are homologous calcium-binding proteins, each present in a variety of neurons in the brain. Their distributions in the rat brain have been compared at the cellular level to determine whether they tend to occur in the same or in different cells, and to determine whether calbindin-positive cells show any common features once crossreaction with calretinin has been eliminated. The results show great heterogeneity. Most cells which contain one of the proteins do not contain the other, but many cells do contain both; even in the ventral cochlear nucleus, where there is abundant calretinin and most calbindin-like immunoreactivity is due to crossreaction, a few cells contain both proteins. In the substantia nigra and ventral tegmental area, many cells are double-positive but some only contain one or the other protein. Only the triangular septal nucleus is uniformly positive for both proteins. Cells which look like local-circuit neurons in many forebrain areas (cortex, hippocampus, olfactory bulb, anterior olfactory nucleus) are exclusively positive for either calretinin or calbindin, in spite of their similar morphology. In the more heterogeneous parts of the brain (including hypothalamus central gray and substantia gelatinosa), there are mixtures of calretinin-positive, calbindin-positive, and double-positive cells. In comparison with previous data on the chick, some aspects of the distributions are conserved, but double-positive cells are more frequent in the rat. The degree of heterogeneity observed, even within comparatively well-defined neuronal populations, makes it difficult to infer in what neuronal properties these proteins could be involved.

Animals↗

Factor analysis of olfactory responses in Drosophila melanogaster enhancer-trap lines as a method for ascertaining common reception components for different odorants.

Olfactory information is transmitted to the brain using combinatorial receptor codes; consequently, a single reception element can be activated by different odorants. Several methods have been applied to describe from a functional point of view those odorants sharing olfactory reception components. A genetic approach in Drosophila melanogaster used correlation between behavioral responses to different odorants for deducing common olfactory pathway-genes. A factor analysis applied to behavioral responses to five odorants of 27 antennal enhancer-trap lines revealed three components, explaining 82.1% of the total observed variance. A first factor affects simultaneously the response to ethyl acetate, propionaldehyde, and acetone. A second factor was related to responses to ethyl acetate, ethyl alcohol, and acetone, and, finally, the third factor associates responses to acetic acid and ethyl acetate. They contribute by 35.1%, 36.9%, and 28%, respectively, to the explained variance.

Animals↗

Epileptiform activity in the piriform cortex of the in vitro isolated guinea pig brain preparation.

Brief intracerebral injections of bicuculline in the anterior portion of the piriform cortex of the isolated guinea pig brain preparation induce a transient, localized disinhibition confined to the site of drug application. The epileptiform activity generated at the bicuculline focus propagates and induces secondary excitability changes in remote cortical regions within the olfactory lobe. Long lasting changes in synaptic potentials were observed in the posterior piriform cortex, where long-projective cortico-cortical fibers originating from the site of bicuculline injection terminate. The activation of rhythmic, transient afterdischarges at the bicuculline focus was critical for the development of persistent 'epileptiform' associative synaptic potentials in remote cortical regions. When transient afterdischarges were prevented, epileptiform associative potentials in the posterior piriform cortex appeared only transiently and vanished within 20 min. The persistent secondary changes in synaptic excitability that occur in cortical regions distant from the acutely-induced primary epileptic focus may represent one of the transition mechanisms toward chronic epileptogenesis.

Animals↗

Ciliary cation conductances in olfactory receptor cells of the clawed toad Xenopus laevis.

One transduction pathway in olfactory receptor neurons is a cascade of receptors, a G-protein, adenylate cyclase, cAMP, and a cyclic nucleotide-activated cation conductance. Here, we show that this conductance is also present in olfactory cells of Xenopus laevis. With optical recordings from the cell's dendritic knob, we show that this conductance, when activated by odors, leads to an increase of intracellular calcium. It is further shown that there is a second cation conductance on the cilia of these cells which is modulated by calcium and can be activated by the application of odorants.

8-Bromo Cyclic Adenosine Monophosphate↗

Sox3 expression identifies neural progenitors in persistent neonatal and adult mouse forebrain germinative zones.

Neural precursors persist throughout life in the rodent forebrain subventricular zone (SVZ) and hippocampal dentate gyrus. The regulation of persistent neural stem cells is poorly understood, in part because of the lack of neural progenitor markers. The Sox B1 subfamily of HMG-box transcription factors (Sox1-3) is expressed by precursors in the embryonic nervous system, where these factors maintain neural progenitors in an undifferentiated state while suppressing neuronal differentiation. Sox2 expression persists in germinative zones of the adult rodent brain, but Sox3 expression in the postnatal brain remains largely unexplored. Here we examine Sox3 expression in the neonatal and adult mouse brain to gain insight into its potential involvement in regulating persistent neural stem cells and neurogenesis. We also investigate Sox3 expression during expansion and neural differentiation of postnatal mouse SVZ neural stem cell and human embryonic stem cell (hESC) cultures. We find that Sox3 is expressed transiently by proliferating and differentiating neural progenitors in the SVZ-olfactory bulb pathway and dentate gyrus. Sox3 immunoreactivity also persists in specific postmitotic neuronal populations. In vitro, high Sox3 protein expression levels in undifferentiated, SVZ-derived neurospheres decline markedly with differentiation. Sox3 immunoreactivity in hESCs appears upon differentiation to neural progenitors and then decreases as cells differentiate further into neurons. These findings suggest that Sox3 labels specific stages of hESC-derived and murine neonatal and adult neural progenitors and are consistent with a role for Sox3 in neural stem cell maintenance. Persistent Sox3 expression in some mature neuronal populations suggests additional undefined roles for Sox3 in neuronal function.

Animals↗

Complementary postsynaptic activity patterns elicited in olfactory bulb by stimulation of mitral/tufted and centrifugal fiber inputs to granule cells.

Main olfactory bulb (MOB) granule cells receive spatially segregated glutamatergic synaptic inputs from the dendrites of mitral/tufted cells as well as from the axons of centrifugal fibers (CFFs) originating in olfactory cortical areas. Dendrodendritic synapses from mitral/tufted cells occur on granule cell distal dendrites in the external plexiform layer (EPL), whereas CFFs preferentially target the somata/proximal dendrites of granule cells in the granule cell layer (GCL). In the present study, tract tracing, and recordings of field potentials and voltage-sensitive dye optical signals were used to map activity patterns elicited by activation of these two inputs to granule cells in mouse olfactory bulb slices. Stimulation of the lateral olfactory tract (LOT) produced a negative field potential in the EPL and a positivity in the GCL. CFF stimulation produced field potentials of opposite polarity in the EPL and GCL to those elicited by LOT. LOT-evoked optical signals appeared in the EPL and spread subsequently to deeper layers, whereas CFF-evoked responses appeared in the GCL and then spread superficially. Evoked responses were reduced by N-methyl-d-aspartate (NMDA) receptor antagonists and completely suppressed by AMPA receptor antagonists. Reduction of extracellular Mg(2+) enhanced the strength and spatiotemporal extent of the evoked responses. These and additional findings indicate that LOT- and CFF-evoked field potentials and optical signals reflect postsynaptic activity in granule cells, with moderate NMDA and dominant AMPA receptor components. Taken together, these results demonstrate that LOT and CFF stimulation in MOB slices selectively activate glutamatergic inputs to the distal dendrites versus somata/proximal dendrites of granule cells.

Action Potentials↗

[Structural organization and connections of cell ensembles of the olfactory bulb of the cat brain].

The cat cerebral olfactory tubercle (OT) includes into its composition certain cellular ensembles distinguished by means of morphological criteria. The ensembles consist of three cellular components: clusters of granular neurons (islands of Calleja), pyramid-like neurons of the layer II, situating along the periphery of the islands, and groupings made by polygonal and spindle-like neurons of the layer III. Morphometrical analysis of every of the three cellular complex components has been carried out. About 7-10 islands of Calleja are situated on the OT territory, which makes nearly 75% of the whole surface of the OT. Neuronal composition of the cellular ensembles has been studied by Golgi method. Varieties of long and short axonal neurons, included into the ensemble composition, have been characterized. Presence of projections of the macrocellular neurons of the layer III (a part of the third component of the ensemble) has been revealed in the posterior part of the lateral hypothalamus and in the field of Forel H1. Possible role of the cellular ensembles is discussed for ensuring various functions of the OT.

Animals↗

The topographical organization of neurons in the rat medial frontal, insular and olfactory cortex projecting to the solitary nucleus, olfactory bulb, periaqueductal gray and superior colliculus.

In 19 rats two different retrograde tracers (Fast Blue, Diamidino Yellow, Rhodamine-labeled latex microspheres, or wheat germ agglutinin conjugated with HRP) were injected into the solitary nucleus (NTS) and either the olfactory bulb (OB), periaqueductal gray (PAG) or superior colliculus (SC). The pattern of retrogradely labeled neurons in the medial frontal, insular and olfactory cortices was examined to determine the topographical organization of the cell populations projecting to these subcortical targets and the extent to which they overlapped. In the medial frontal cortex (MFC) SC projections originated most dorsally, while NTS and OB projections originated most ventrally and exhibited slight overlap. PAG projections originated from virtually the entire MFC and overlapped with cells projecting to the OB, NTS and SC. These results are consistent with the role of dorsal MFC as the rat's frontal eye field and the ventral MFC as a visceral motor area. Laterally, in the insular cortex there was virtually complete overlap between cells projecting to the NTS and PAG. The extensive overlap of PAG projections with NTS projections medially and laterally and with SC projections medially suggests the PAG is involved in a variety of brain visceral and somatic functions. In the piriform cortex there was overlap between cells projecting to the OB and cells projecting to the SC; the cells projecting to the SC were located in the endopiriform nucleus, and may provide a substrate for orienting responses to odors.

Animals↗

Calretinin expression in specific neuronal systems in the brain of an advanced teleost, the grey mullet (Chelon labrosus).

The distribution of calretinin (CR) in the brain of an "advanced" teleost, the grey mullet, was studied by using immunoblotting and immunocytochemical techniques. In immunoblots of protein extracts of rat and mullet brains, the CR antibody stained a single band of about 29 kDa. CR immunoreactivity was observed in specific neuronal populations of all brain regions. The primary olfactory system, the optic nerve fibers, and some sensory fibers of other cranial nerves exhibited strong CR immunoreactivity. In the forebrain, the CR-immunoreactive (CR-ir) populations were scarce in the telencephalon and hypophysiotrofic hypothalamus, but numerous in many specialized nuclei of the diencephalon (preglomerulosus complex, nucleus glomerulosus, anterior glomerular nucleus, nucleus diffusus) and pretectum (parvocellular and magnocellular superficial pretectal nuclei, central pretectal nucleus), which are related to sensory systems. The two main forebrain bundles, medial and lateral, contained numerous CR-ir fibers. The midbrain sensory centers (optic tectum and torus semicircularis) exhibited numerous CR-ir cells and fibers. Likewise, the secondary gustatory nucleus of the isthmus is one of the nuclei exhibiting more intense CR immunoreactivity. Characteristically, the efferent cerebellar system (eurydendroid cells and brachium conjunctivum) and some afferent cerebellar fibers were CR-ir. In the medulla oblongata, a number of reticular cells, the inferior olive, and the magnocellular octaval nucleus exhibited CR immunoreactivity. CR-ir motoneurons were also observed in the spinal cord and in the oculomotor nucleus. Together with results obtained in other vertebrates, present results suggest that neural systems using calretinin to maintain intracellular calcium concentration have been rather well conserved during vertebrate evolution.

Animals↗

Smelling of odorous sex hormone-like compounds causes sex-differentiated hypothalamic activations in humans.

The anatomical pathways for processing of odorous stimuli include the olfactory nerve projection to the olfactory bulb, the trigeminal nerve projection to somatosensory and insular cortex, and the projection from the accessory olfactory bulb to the hypothalamus. In the majority of tetrapods, the sex-specific effects of pheromones on reproductive behavior is mediated via the hypothalamic projection. However, the existence of this projection in humans has been regarded as improbable because humans lack a discernable accessory olfactory bulb. Here, we show that women smelling an androgen-like compound activate the hypothalamus, with the center of gravity in the preoptic and ventromedial nuclei. Men, in contrast, activate the hypothalamus (center of gravity in paraventricular and dorsomedial nuclei) when smelling an estrogen-like substance. This sex-dissociated hypothalamic activation suggests a potential physiological substrate for a sex-differentiated behavioral response in humans.

Adult↗

Similarity of granular-induced inhibitory periods in pairs of neighboring mitral/tufted cells.

1. Neighboring mitral/tufted cells have been previously shown to present temporal correlations of their firings related to the respiratory rhythm, particularly under odor stimulation. This occurs despite the existence of a powerful inhibitory control exerted by granule cells onto mitral/tufted cells. In the present study, we hypothesized that neighboring mitral cells can present granular induced inhibitory periods with similar latencies and durations and that such a similarity would preserve them from a possible suppression of their temporal correlations. 2. To test this hypothesis, we analyzed the latencies and durations of the inhibitory periods induced by granular activation in pairs of simultaneously recorded neighboring mitral cells. The activation of granule cells was achieved by electrical stimulation of the different pathways known to directly activate granule cells [lateral olfactory tract (LOT), anterior limb of the anterior commissure (AC), and piriform cortex (PC)]. Data from this group were compared with those of a control group composed of distant cells also recorded simultaneously. 3. Results first show that the latencies to onset of inhibition or to recovery were more frequently similar in neighboring cells than in control cells and that this similarity was enhanced by odor stimulation. Second, the probability that two cells exhibit similar inhibitory periods (i.e., similar latencies to both onset and to recovery) in response to electrical stimulation of LOT, AC, or PC was significantly higher in neighboring than in control cells. Third, only neighboring cells were found to present similar inhibitory periods in response to the stimulation of all of the three structures. 4. Granular activation was also found to modify the temporal patterns of individual mitral cells. However, although these patterns were not systematically modified similarly in neighboring mitral cells, they remained perfectly synchronized with zero delay if they were already synchronous without electrical stimulation. On the contrary, if patterns were spontaneously uncorrelated, electrical stimulation never produced a synchronization of their firings, even if their temporal relationships could be profoundly modified. 5. These results show that neighboring mitral cells can receive granular-induced inhibition with similar latencies and durations with a probability much higher than control cells. Such similarities allow neighboring mitral cells to preserve their temporal correlation despite the powerful inhibitory input from granule cells. Functional hypotheses about the role of the cortical feedback projections onto the bulb are discussed.

Animals↗