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Monosynaptic and disynaptic activation of pyriform cortex neurons by synchronous lateral olfactory tract volleys in the rabbit.

To elucidate the organization of synaptic inputs to pyriform cortex neurons, intracellular and extracellular responses of single units were analyzed in urethane-anesthetized rabbits. The lateral olfactory tract (LOT) or the olfactory bulb (OB) was electrically stimulated. Intracellular recordings revealed two types of cells (type I and type II cells), according to the types of EPSP evoked by the LOT or OB shock. The EPSP in the type I cells had shorter latencies (0.0 to 0.9 ms) from the onset of the component 2 (C2) wave of the field potential (which signals the onset of the synaptic depolarization of the apical dendrites of the pyramidal cells in the PC), and that in the type II cells had longer latencies (1.0 to 6.0 ms). A conditioning LOT or OB shock did not suppress the testing EPSP in the type I cells, whereas the conditioning stimulation greatly suppressed the testing EPSP in most of the type II cells. Extracellular recordings from units responding synaptically to the LOT or OB shock revealed a group of units which had short latencies (0.7 to 1.9 ms) of spike discharges. Those units, which were likely to be the same cells as the type I cells, are believed to mediate excitatory synaptic inputs to the type II cells. On the basis of these results, we concluded that type I cells are monosynaptically activated by LOT volleys, whereas type II cells are activated di- or polysynaptically by way of a relay from type I cells. The type I cells were recorded in both the superficial and the deep parts of the pyriform cortex, although they were recorded more frequently in the superficial part. On the other hand, most of the type II cells were recorded in the deep part of the PC. These results support and extend the previous model, in which the monosynaptically activated superficial pyramidal cells give rise to excitatory inputs to other pyramidal cells and neurons in deep layers.

Animals↗

Cellular events induced in the molecular layer of the piriform cortex by ablation of the olfactory bulb in the mouse.

Cellular events associated with degeneration of the projection of the olfactory bulb to the molecular layer of the piriform cortex of the mouse have been studied with rapid-Golgi and Fink-Heimer impregnations and with the electron microscope. Four classes of axon terminals: s-1, s-d, f-1, and f-d, are differentiated on the basis of whether the synaptic vesicles are spherical or flattened and whether the axoplasm is lightly or darkly stained. The majority of s-1 terminals, the predominant class in sublamina Ia of the molecular layer, degenerate after bulb ablation. Degeneration of axon terminals is associated with dilation and, eventually, degeneration of segments of dendrites in Ia. Both s-1 and s-d terminals contribute to a partial reconstitution of the neuropil of Ia during the weeks after bulb ablation.

Animals↗

Binary expression of olfactory bulb-protein tyrosine phosphatase in rat central nervous system: developmental gene regulation in neonate cerebral cortex and constitutive expression in olfactory-rhinencephalon.

Olfactory bulb-protein tyrosine phosphatase (OB-PTP) is a receptor type PTPase dominantly expressed in olfactory bulb. Previously, we isolated and molecularly cloned a rat OB-PTP cDNA from an embryonal brain cDNA library. In the present study, we investigated its temporal and spatial gene expression by Northern blot and in situ hybridization analysis. The expression of OB-PTP gene was firstly detected in day 16 post coitum embryo and significantly increased during the late-gestational stage, attaining the highest level in the first week of neonate. The OB-PTP transcript was then down-regulated postnatally and was detected barely in an adult brain. In situ hybridization analysis showed that the transcript was characteristically localized in the postmitotic neurons of cerebral cortex and subcortical structures, and was down-regulated by day 28 when the cortical and subcortical structures have been organized. In the olfactory-rhinencephalon system including olfactory bulb and piriform cortex, the OB-PTP was preferentially expressed in the postmitotic neurons, and in contrast continuously expressed in the matured brain. Based on the evidence that DPTP10D, the Drosophila homolog of OB-PTP, is localized in the axons of specific pioneer neurons in Drosophila embryo, the OB-PTP is presumably involved in the axonogenesis of cortical and subcortical neurons as well as olfactory neurons in mammalian central nervous system. The biological significance of transcriptional regulation in olfactory system is discussed in terms of continuous axonal connections by regenerating olfactory neurons.

Animals↗

GABA and opioid binding distribution in the brain of the seizure-resistant Proechimys guyannensis: an autoradiography study.

Proechimys guyannensis rodents present resistance to epilepsy. Autoradiography was used to map GABA(A) ((3)H-Muscimol), benzodiazepine ((3)H-Flunitrazepam), mu ((3)H-DAMGO), and delta ((3)H-DPDPE) opioid receptor binding in adult Proechimys guyannensis brain under normal conditions. Results were compared with values obtained from adult Wistar rats. Proechimys presented reduced (3)H-Muscimol binding in several cortices, thalamus, medial amygdala nucleus, and dorsal dentate gyrus. (3)H-Flunitrazepam binding was reduced in periaqueductal gray, frontal and entorhinal cortices, and enhanced in piriform cortex and ventral CA2 field of Ammons horn. Concerning (3)H-DAMGO binding, high values were found in several cortices, medial amygdala nucleus, dorsal dentate gyrus, and periaqueductal gray, whereas reduced binding was detected in anterior olfactory tubercle, cingulated cortex, thalamus, basolateral amygdala, substantia nigra, and dorsal and ventral CA fields. High (3)H-DPDPE binding was noticed in CA1 field from dorsal hippocampus, while reduced values were found in cingulate cortex, olfactory tubercle, thalamus, and substantia nigra. These findings provide the first description of receptor binding distribution of the Proechimys brain and suggest natural endogenous anticonvulsant mechanisms of theses rodents under normal conditions.

Animals↗

Light-microscopic immunocytochemical localization of tyrosine hydroxylase in prenatal rat brain. II. Late ontogeny.

The immunocytochemical localization of tyrosine hydroxylase is examined at embryonic (E) days 18 and 21 in rat brain in order to determine changes in the distribution and cytology of neurons showing immunoreactivity for the enzyme during late prenatal development. As compared with earlier stages of development, the distribution and morphology of the tyrosine hydroxylase-containing neurons at E18 and E21 more closely resemble catecholaminergic neurons in the adult brain. The changes occurring from the early to the late prenatal stages of development appear to be the result of an increase in number of cells and continued aggregation and migration of the labeled neurons. The major differences in the distribution of labeled perikarya between E18 and E21 are in the olfactory bulb and cerebral cortex. In the olfactory bulb, tyrosine hydroxylase-containing neurons are not detected until E21. In contrast in the cerebral cortex, a few neurons are transiently labeled for the enzyme at E18, but are not detected at E21 and have not been reported in the adult brain. The most striking change in the tyrosine-hydroxylase labeled structures in the late prenatal period is the increase in detectable immunoreactivity in bundles of axons and in terminal aborizations. The orderly appearance of tyrosine hydroxylase-labeled axons in the neostriatum and cortex are discussed in relation to the formation of these two contrasting regions innervated by catecholaminergic neurons.

Animals↗

Regional differences in glial-derived factors that promote dendritic outgrowth from mouse cortical neurons in vitro.

To determine whether glia from different CNS regions differ in their ability to support axons or dendrites, embryonic (E18) mouse cortical neurons were cocultured with early postnatal (P4) rat astroglial derived from cortex, retina, olfactory bulb, mesencephalon, striatum, and spinal cord. After 5 d in vitro, axon and dendrite outgrowth from isolated neurons was quantified with double-labeling immunohistochemical techniques. Whereas axonal growth was similar on the various monolayers, total primary dendritic outgrowth was nearly threefold greater on glia derived from the cortex, retina, and olfactory bulb than on glia derived from mesencephalon, striatum, or spinal cord. This effect was principally on the number of primary dendrites rather than the elongation of individual dendrites. Similar morphological differences were observed when cortical neurons were grown on polylysine in a noncontact coculture system with glia continuously conditioning the media. This selective promotion of dendrite growth was independent of neuron survival. These results indicate that there are regional differences in the ability of CNS glia to support dendritic growth and that this effect is due, in part, to release of a diffusable factor.

Animals↗

Developmental expression of 5-HT 5A receptor mRNA in the rat brain.

In the central nervous system, serotonin (5-HT) may function as a mitogen as well as a neurotransmitter; and its early appearance suggests a potential role in development. The present experiments were done to determine the localization of the mRNA coding for the 5-HT 5A receptor during development of the rat brain. 5-HT 5A gene transcription was assessed by in situ hybridization, from E18 and during postnatal (PN) development. An intense signal of 5-HT 5A mRNA was found in the cerebral cortex and olfactory nucleus at E18, PN0 and PN5. A sharp decrease at PN11 was followed by an increase until reaching the adult level in the cerebral cortex; whereas in the olfactory nucleus, transcription remained weak. In contrast, in the hippocampal formation the signal was weak in the CA1, CA2 and CA3 regions at E18 and PO; increased at P5 and then decreased at P11 before attaining the adult level. We conclude that the gene coding for the 5-HT 5A receptor is already active in the embryonic rat brain and is differentially expressed during development.

Animals↗

The zinc iodide-osmium (ZIO) reaction in the central nervous system: localization and relations to functional activity.

The fine structural characteristics of ZIO reaction was studied in the cerebral and cerebellar cortex and olfactory bulb of the rat and in synaptosomes prepared from rat cerebellar cortex. It was concluded that: 1. Organelles of different nerve cell types exhibit different ZIO reactions provided that the impregnation was carried out under standardized conditions. 2. 6...10 times more synaptic vesicles were stained by ZIO in the inhibitory terminals than in the excitatory ones. 3. ZIO positivity was found in all types of synaptosomes prepared from cerebral cortex. Following electrical or chemical (KCl) depolarization there was a decrease in the number of ZIO positive synaptic vesicles, which decrease was directly proportional to the parameters of stimulation. 4. By x-ray microanalysis Os, Zn and Ca were consistently detected in the ZIO precipitates. Iodine, however, could not always be found. After stimulation the presence of Ca was observed even in those synaptosomes in which the ZIO reaction product was absent. 5. On the basis of the staining characteristics the reaction, under standard conditions, can reflect certain functional states of the nerve terminals.

Animals↗

Structure-activity requirements of bombesin for gastrin-releasing peptide- and neuromedin B-preferring bombesin receptors in rat brain.

The pharmacological profile of [125I][Tyr4]bombesin binding to gastrin-releasing peptide- and neuromedin B-preferring sites has been investigated in rat cerebral cortex and olfactory bulb membranes, respectively. [125I][Tyr4]bombesin specific binding to cerebral cortex membranes was displayed biphasically by gastrin releasing peptide and [D-Phe6]bombesin-(6-13)-ethyl amide. In the presence of 10 mM neuromedin B, displacement curves for bombesin-related peptides were monophasic with gastrin releasing peptide displaying approximately 100-fold higher affinity than neuromedin B. In olfactory bulb membranes, [125I][Tyr4]bombesin binding was also displaced biphasically by gastrin releasing peptide, [D-Phe6]bombesin-(6-13)-ethyl amide and neuromedin B. In the presence of 10 microM [D-Phe6]bombesin-(6-13)-ethyl ester, displacement curves were monophasic with neuromedin B possessing approximately 10-fold higher affinity than gastrin-releasing peptide. Under these conditions, successive deletion of N-terminal amino acids from bombesin-(1-14) was well tolerated at both sites, with little loss in affinity up to bombesin-(5-14). A 5- to 10-fold drop in affinity was observed at both sites with bombesin-(6-14), whilst the octapeptide acetyl-bombesin-(7-14) displayed similar affinities to bombesin-(1-14). Bombesin-(8-14), -(9-14) and -(10-14) were essentially inactive (IC50 > 10 microM). C-terminal deletion of Met24 (bombesin-(1-13)) resulted in 100-fold loss of affinity at the gastrin-releasing peptide site and complete loss of affinity at the neuromedin B site. Fragments smaller than bombesin-(1-13) were virtually inactive at either site.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Localization of atrophy-prone areas in the aging mouse brain: comparison between the brain atrophy model SAM-P/10 and the normal control SAM-R/1.

Mouse inbred strain "SAM-P/10" (Senescence Accelerated Mouse) is a model of age-related brain atrophy. In this strain there is an earlier and more severe age-related deterioration in the conditional avoidance learning than the normal control inbred SAM-R/1 strain. The present study analysed age-related changes in brain area size using a computerized morphometric method. The region most vulnerable to age-related atrophy in SAM-P/10 was the frontal region of the cerebral cortex, including the prefrontal cortex. Other neocortical regions underwent diffuse atrophy. Posterior piriform cortex, entorhinal cortex, anterior olfactory nucleus, amygdala, caudate-putamen, nucleus accumbens and cerebellar cortex were atrophy-prone regions. The septum also underwent atrophy but other basal forebrain structures were intact. The hippocampus, diencephalon and brainstem structures showed no atrophic change. White matter structures did not change in size with aging except for the forceps minor of the corpus callosum, which showed age-related atrophy. On the contrary, SAM-R/1 showed a significant age-related atrophy only in a restricted part of the cerebral cortex, mainly in the parietal region. Other cortical regions, subcortical structures, diencephalon, brainstem structures, cerebellum and white matter were atrophy-resistant in SAM-R/1. The prefrontal cortex, entorhinal cortex, piriform cortex and striatum are closely interconnected and also connect with the amygdala which plays a key role in conditioning in the rodent. Age-related atrophy in all these structures in SAM-P/10 presumably accounts for the age-related deficits in conditional avoidance learning in this strain of mouse. Comparison between SAM-P/10 and SAM-R/1 or other well-known rodents indicates that SAM-P/10 is a unique rodent that spontaneously and rapidly develops progressive generalized cerebral atrophy, which is considered to be a pathological process rather than an accelerated aging process.

Aging↗

Expression of stef, an activator of Rac1, correlates with the stages of neuronal morphological development in the mouse brain.

STEF (Sif- and Tiam1-like exchange factor), a guanine nucleotide exchange factor, was identified as a candidate molecule in regulation of neural development. The STEF gene product specifically activates Rac1, a member of the Rho-like small G proteins. Here we report the detailed examination of the expression profile of the stef gene in the mouse brain. In situ hybridization revealed that the stef gene was expressed in a stage- and region-specific manner in the mouse brain; it was expressed during certain developmental stages in the cerebral cortex, the olfactory bulb, the rostral migratory pathway (RMP) and the hippocampus. In the cerebral cortex, stef transcripts were detected in migrating cells in the intermediate zone as well as neurons in the cortical plate. While the expression in the cerebral cortex was reduced at adult stages, considerable expression was found to be maintained in other regions (RMP, olfactory bulb, hippocampal formation), which are the tissues where neurons continue to undergo morphological remodeling including cellular migration, neurite extension and synapse formation even in adults. Thus, stef gene expression appears to correspond to neuronal morphological changes.

Animals↗

Olfactory bulb connections with basal rhinencephalon in the ferret: an evoked potential and neuroanatomical study.

The lateral olfactory tract (LOT) of the ferret has been shown to project to most of the pyriform lobe, as in the cat. Only a small medio-posterior region of the pyriform cortex (the angular cortex), which has a distinctly different appearance in Nissl stained sections, proves to be devoid of olfactory connections. Despite the fact that sub-areas can be recognized within olfactory cortex, there is an underlying constancy in design throughout. Latency measurements indicate that fine collateral branches of theLOT reach the posterior olfactory cortex, whereas mainly larger diameter (faster conducting) fibres comprise the contribution to anterior olfactory cortex. Thus in the present context it is sufficient to recognise just an anterior and posterior subdivisionof olfactory cortex, contingent on the above criterion and correlated with cytoarchitectural features, chiefly variations in cortical layers II and III of the regions concerned. Pyriform cortex directly in receipt of LOT fibres, and associated deeper cortical zones, generated potentials in the olfactory bulbs which can only be attributed to centrifugal input, thus further substantiating an olfactory role for this cortex. In addition, non-cortical regions in receipt of LOT fibres, namely the anterior olfactory nucleus, olfactory tubercle, cortical amygdaloid nucleus and nucleus of the lateral olfactory tract, also contribute centrifugal input to the bulbs. All these regions are thereby capable of providing a rather direct feedback on olfactory bulb activity. The exact pathways concerned have yet to be determined.

Animals↗

Developmental expression of neural cell adhesion molecules in the mouse neocortex and olfactory bulb.

Polyclonal antibodies to N-CAM and L1 and monoclonal antibodies to epitopes of N-CAM (designated 12F11, 8A2, and 12F8) were used to investigate the spatial and temporal distribution of these neural cell adhesion molecules during the development of mouse cortex and olfactory bulb. The aim of the study was to correlate developmental events such as cell migration, dendritic and axonal outgrowth, and synaptogenesis with the appearance and disappearance of specific molecules involved in cell-cell interactions. Western transfer studies indicated that 12F8 antibody recognized polysialic acid found on embryonic N-CAM; 8A2 antibody primarily recognized the 140 kD component of N-CAM while the 12F11 antibody recognized the 180 and the 140 kD forms. The study demonstrates a high degree of cell surface molecular specialization of different compartments in developing neocortex and olfactory bulb. L1 is found on a variety of unmyelinated fiber tracts including thalamocortical fibers, olfactory nerve, and inner plexiform layer of the olfactory bulb. In contrast, N-CAM epitope recognized by 12F11 antibody is present on olfactory nerve fibers but appears later and is much weaker than L1 on thalamocortical fibers and is absent from the olfactory lobe inner plexiform layer. Dendritic regions are best labeled by 12F8 antibody; the epitope becomes faint in adult cortex but remains strongly expressed in olfactory bulb. This study reveals that widespread N-CAM expression in the central nervous system is constituted by a diversity of local expression of different molecular forms of N-CAM; their different anatomical distributions suggest they may each have unique roles.

Aging↗

Differential effects of olfactory bulbectomy on beta-adrenoceptors in rat amygdala, hippocampus and cerebral cortex.

Rats were bilaterally olfactory bulbectomized. At 15 days post-lesion, olfactory bulbectomized (OB) rats exhibited significant deficits in the acquisition of passive avoidance learning compared to sham lesioned rats. beta-Adrenoceptor binding in the amygdala, hippocampus and cerebral cortex was assayed with (-)-[125I]iodocyanopindolol (ICYP). Scatchard analyses revealed no difference between OB and sham rats in maximal binding density (Bmax) in any of the three tissues. However, in the OB rats, the affinity of the beta-adrenoceptor for the ligand was significantly increased in the amygdala and hippocampus but not in the cortex. Bulbectomy did not affect the ratio of beta 1- to beta 2-adrenoceptor subtypes in the three brain tissues. In amygdala and hippocampus but not cerebral cortex, bulbectomy resulted in an increase in the proportion and the affinity of the high-affinity beta-adrenoceptor binding sites for isoproterenol. The affinity of the low-affinity sites in the hippocampus was also increased in the OB rats. The results suggest that olfactory bulbectomy causes supersensitivity of the amygdaloid and hippocampal beta-adrenoceptor by increasing the degree of coupling of the receptor with the stimulatory guanine nucleotide binding protein (Gs protein).

Amygdala↗

Cellular correlates of olfactory learning in the rat piriform cortex.

This review describes research that combines cellular physiology with behavioral neuroscience, to study the cellular mechanisms underlying learning and memory in the mammalian brain. Rats were trained with an olfactory conditioning paradigm, in which they had to memorize odors in order to be rewarded with drinking water. Such training results in rule learning, which enables enhanced acquisition of odor memory. Training results in the following learning-related physiological modifications in intrinsic and synaptic properties in olfactory (piriform) cortex pyramidal neurons: 1. increased neuronal excitability, indicated by reduced afterhyperpolarization, and 2. increased synaptic transmission, indicated by reduced paired-pulse facilitation. These modifications are correlated to enhanced learning capability rather than to storage of memory for specific odors. In addition, using a different paradigm of odor-training, it is shown that NMDA and betra-adrenergic receptors are involved at different stages of long-term memory consolidation.

Animals↗

Cellular and intracellular localization of epsilon-subspecies of protein kinase C in the rat brain; presynaptic localization of the epsilon-subspecies.

The cellular and intracellular localization of the epsilon-subspecies of protein kinase C (PKC) in the rat brain was demonstrated by immunocytochemistry using specific antibodies against epsilon-PKC. The epsilon-PKC-specific immunoreactivity was most abundant in the hippocampal formation, olfactory tubercle and Calleja's islands, was moderate in the cerebral cortex, anterior olfactory nuclei, accumbens nucleus, lateral septal nuclei and caudate-putamen and low in the thalamus and medulla. The epsilon-PKC-immunoreactivity was scanty in the perikarya, except for the pyramidal cells of CA3 region of the hippocampus and the immunoreactivity was mainly present in neuropils and nerve fibers. The distribution of epsilon-PKC immunoreactive neurons was consistent with that obtained by in situ hybridization histochemistry. Electron microscopic observations of the hippocampus revealed that the epsilon-PKC is predominantly present in the cytoplasm of axon and nerve terminals and that this enzyme is associated with mitochondrial membrane and vesicles. These results suggested that epsilon-PKC is probably involved in presynaptic functions in CNS, perhaps even neurotransmitter release.

Amino Acid Sequence↗