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Canine limbic system substance P receptors.

A canine limbic system preparation is utilized as a source of substance P receptor(s) to screen gradient RP-HPLC fractions for the presence of receptoractive-substance P activity, and to quantify endogenous receptoractive-substance p in biological extracts such as human tooth pulp. The binding characteristics, KD = 1.3 nM and Bmax = 11 fmol mg-1 protein, are similar to values obtained from receptors produced from other biological sources such as whole rat brain, minus cerebellum.

Animals↗

[Morphologic image of structures of the limbic system in rats after prolonged administration of antiepileptics].

Morphological changes in the anatomical structures of the limbic system induced by phenytoin and ethosuximide administered in effective doses for 1, 3 and 6 months in rats were analyzed. The clinical symptoms consisted of some vegetative and behavioral disorders, mostly transitory. Differences in gain of body weight depending on the sex of the animal and the drug administered were observed. The morphological changes in the parenchymal and mesenchymal nervous tissue elements were not specific with regard to their topography or to the type of pathological process. Some morphological differences depending on the drug and its period of administration were observed only in rats treated for 1 and 3 months. The neuropathological picture in rats treated with both drugs for 6 months showed a great similarity. The pathological process in the ganglionic cells had the character of degeneration. Morphological changes in the myelin sheaths were due to edema which appeared to be vasculogenic and situated in the white matter. Focal and diffuse proliferation of cellular glia appeared after administration of both drugs for 1 and 3 months while in those treated for 6 months the degenerative changes were seen. The anatomo-comparative study of the neuropathological picture in the rats treated with both drugs and the morphological picture of the limbic system structures in patients with chronic epilepsy indicates that the drugs examined could play some role in the pathogenesis of the limbic system lesions encountered in epileptics.

Animals↗

Analysis of facial displays and verbal report to assess subjective state in the non-invasive detection of limbic system activation by procaine hydrochloride.

The problem of scalp EEG as a measure of cortical or subcortical activity is particularly relevant to complex partial seizures as the abnormal discharging is frequently limbic in origin [14, 30]. Livingston [38] has suggested that administration of intravenous procaine as a limbic activator and cortical suppressor would be of utility in diagnosing limbic involvement in complex partial seizures. While there is considerable evidence derived from experimental animal models that procaine hydrochloride is a limbic system activator that acts preferentially on subcortical epileptic foci at lower doses than on less active epileptic foci or non-epileptic tissue [2, 4], it was necessary to demonstrate that procaine activates the human limbic system. The non-invasive approach taken in the present study was to compare the published effects of direct electrical stimulation of the human limbic system [31] to the behavioural and subjects effects of intravenous procaine administration. The areas in which we obtained the most robust procaine effects (hallucinations, emotions and alimentary sensations) were also Halgren et al.'s [31] most repeatable effects. The correspondence between electrical stimulation effects and procaine administration effects was striking - with verbal report by patients matching exactly in many instances. Furthermore, analysis of facial displays proved useful in providing access to subjects state fluctuations which would otherwise have gone undetected. The data provide strong evidence that procaine hydrochloride can be used as a human limbic system activator. Future research will investigate the clinical and diagnostic significance of differential response to procaine.

Diazepam↗

Developmental and regeneration-associated regulation of the limbic system associated membrane protein in explant cultures of the rat brain.

In the present study we have examined the topographic and temporal patterns of expression of the limbic system associated membrane protein by light and electron microscopic immunocytochemistry in organotypic cultures of the rat brain. The regional, cellular and subcellular distribution of staining in young cultures was similar to that in the intact brain of corresponding age. Since the tissue in vitro is isolated both from afferents and targets, short-term protein expression appears to be regulated by factors intrinsic to the neuron. In culture, the protein was present on the surface of neurons which are physiologically interconnected, such as neurons belonging to the septohippocampal system (cholinergic neurons in the septum and pyramidal and granule cells in the hippocampus). It was also present on the surface of axons and growth cones during process outgrowth. Thus, the limbic system associated membrane protein is expressed in an appropriate spatial and temporal pattern for mediating interactions between growing axons and their targets. The expression of the protein in culture showed some important differences as compared to the intact brain. With increasing age, there was an increasing scattering and disappearance of immunoreactivity in cultures fixed with paraformaldehyde/glutaraldehyde. The decreased immunoreactivity in aged cultures does not appear to reflect decreased protein synthesis, because unfixed and acetone-fixed explants continued to show immunostaining. Furthermore, dot-blot assays showed similar amounts of immunoreactivity in culture as in the intact brain of corresponding age. Thus, the age-dependent decrease of immunoreactivity may reflect altered insertion of the protein into the membrane or a modification of the epitope recognized by the antibody. There was a rapid increase (within 1 hour) of immunostaining on the surface of sprouting processes following mechanical lesion of mature, unstained axons. The altered distribution after tissue injury could be a means of ensuring specificity of connectivity during nerve fiber regeneration. On the basis of the reported findings, we suggest that system-specific membrane proteins, including the limbic system associated membrane protein, may mediate the formation of specific connections in the brain. Furthermore, we suggest that the reinnervation processes taking place after central nervous system injury may exhibit a similar molecular basis to the development of neural pathways.

Animals↗

The effect of drugs of abuse on NMDAR1 receptor expression in the rat limbic system.

An increasing body of evidence points to the role of N-methyl-D-aspartate (NMDA) receptors in the limbic system in the mechanism of drug dependence. We studied the influence of acute and repeated morphine (20 mg/kg i.p. or increasing dose for 10 days) and cocaine (3x20 mg/kg i.p. per day at hourly intervals, for 1 or 5 days) administration on the expression of glutamate NMDA receptor subunit 1 (NMDAR1) in the central and basolateral nuclei of the rat amygdala and hippocampal formation. Acute or chronic morphine and cocaine administration increased NMDAR1 mRNA level in the central and basolateral nuclei of the amygdala; morphine did so 3 h after the last dose and 48 h after withdrawal, cocaine 3 h after acute and last chronic dose. Morphine did not change the NMDAR1 mRNA level in the hippocampal formation, but chronic cocaine did decrease it in the dentate gyrus only. Our study suggests a possible link between the expression of NMDAR1 and changes in limbic system neuronal activity and behaviour after administration of morphine and cocaine. In summary, the present study demonstrated that morphine and cocaine influenced the expression of NMDAR1 in the structure of the limbic system which could be involved in dependence phenomena.

Amygdala↗

Increased limbic system symptomatology and sensitizability of young adults with chemical and noise sensitivities.

We previously hypothesized that individual differences in (a) limbic system reactivity and (b) central nervous system sensitizability underlie vulnerability to environmental stimuli, not only in the controversial clinical condition multiple chemical sensitivity (MCS), but also in the general population. Earlier research has shown overlaps in the characteristics of persons who report noise and air pollutant sensitivities. This study assessed questionnaire responses of 897 young adult college students who reported high versus low frequency of illness from several environmental chemical odors and concomitantly high versus low sensitivity to environmental noise. Subjects who reported increased rates of illness from chemical odors with or without noise sensitivity scored significantly higher (P < 0.0001) on a measure of limbic system symptomatology derived from ictal sensory, somatic, mnemonic, and behavioral manifestations of temporal lobe epilepsy. The group rating high both for illness from chemicals and for noise sensitivity had characteristics predictive of heightened sensitizability from the animal research on time-dependent sensitization (progressive response amplification to repeated, intermittent stimuli over time): i.e., higher female to male ratio (gender risk factor), increased rates of drug abuse problems in blood relatives (genetic risk factor), trait shyness (hyperreactivity to novelty), and increased carbohydrate craving. Despite the increased family histories of drug abuse and levels of personal anxiety and depression, the chemical- and noise-sensitive group reported the lowest rates of current smoking or personal drug abuse problems and the highest frequency of illness from drinking a small amount of alcohol. Taken together, the findings suggest that limbic system dysfunction associates more with chemical than with noise sensitivity; that individuals with both chemical and noise sensitivity may be the most sensitizable subset of the population for prospective studies, and that, in their substance use patterns, young adults with both chemical and noise sensitivity are more similar to MCS patients than are their peers with chemical or noise sensitivity alone.

Adolescent↗

The tegmental pedunculopontine nucleus: a brain-stem output of the limbic system critical for the conditioned place preferences produced by morphine and amphetamine.

The potent reinforcing properties of psychoactive drugs have been attributed to the activation of motivational processes localized to the limbic system. We investigated the role of 2 specific outputs of the forebrain limbic system, the tegmental pedunculopontine nucleus (TPP) and the periacqueductal gray (PAG) of the pons-midbrain, in the positive motivational effects of morphine and amphetamine. We now report that the TPP, but not the PAG nor other nearby regions, is a critical site in the neural system subserving the rewarding effects of both opiates and stimulants. Bilateral ibotenic acid lesions of the TPP blocked the positive reinforcing effects of both morphine and amphetamine in naive rats as measured in a conditioned place preference paradigm. However, TPP lesioned animals were still capable of acquiring a conditioned place preference to an environment paired with the peripheral opiate antagonist methylnaltrexone. This suggested that TPP lesions did not cause nonspecific deficits in the basic learning mechanisms underlying conditioned place preferences. Furthermore, while the TPP was critical for the acquisition of a conditioned preference to an environment paired with morphine in naive rats, rats that had acquired a morphine conditioned place preference prior to the lesions were capable of retaining and demonstrating these place preferences after lesions of the TPP. This again demonstrates that TPP lesions are producing an unconditioned deficit in motivation rather than a deficit in learning or memory. Finally, direct comparisons of the place preference data of individual animals with their correspondent TPP lesion sites indicated that the most effective lesions overlapped to a greater degree TPP perikarya with descending, rather than ascending, axons. This suggests that motivational information generated by drug stimuli acting at "upstream" neural structures flows in a descending direction through the TPP region of the brain stem. These results suggest that opiates and stimulants must ultimately activate a single brain-stem substrate in order to have a positive motivational impact. It is hypothesized that the neural circuits mediating the rewarding effects of drug stimuli acting at forebrain sites exit the limbic system in the TPP region of the brain stem, where motivation may ultimately influence or be isomorphic with the elicitation of motor responses subserving approach and exploration.

Amphetamine↗

Distribution of 1,25-dihydroxyvitamin D3 receptor immunoreactivity in the limbic system of the rat.

We used immunocytochemistry to obtain a complete cellular and subcellular mapping of the 1,25-dihydroxyvitamin D3 receptor protein (VDR) in the rat limbic system. We observed specific VDR immunostaining in the nucleus as well as in the perinuclear cytoplasm of neuronal cells. The limbic system consists of a variety of neuronal structures, and is known to have influence on memory, behavior, emotions and reproduction. In the hippocampal formation, we found strong nuclear staining as well as less distinguished cytoplasmic VDR staining in CA1, CA3 and CA4. The CA2 area showed a unique cytoplasmic predominance of VDR. The amygdala was found to exhibit specific patterns of VDR distribution in the various regions of the nucleus. We observed distinct differences of VDR localization within the limbic preoptic areas of the hypothalamus. Further parts of the brain we analyzed included the mammillary bodies, the indusium griseum and the cingulate cortex. The subcellular distribution of VDR in regions of the limbic system suggests a specific functional role of the receptor protein and indicates a role for calcitriol as a neuroactive steroid.

Amygdala↗

Corpus callosum and limbic system: neuroanatomic MR evaluation of developmental anomalies.

Agenesis of the corpus callosum is a complex malformation of the brain that has been associated with varying degrees of limbic system maldevelopment. We retrospectively reviewed the records of 11 patients with callosal agenesis (seven total, four partial) who underwent magnetic resonance (MR) imaging, with particular attention to the associated malformations of the limbic system. Comparison was made with selected images from MR examinations of healthy volunteers and with necropsy specimens from other patients with callosal agenesis. Ten of 11 patients demonstrated limbic anomalies (severe motion artifact precluded evaluation of these structures in one patient). MR depicted not only the abnormalities intrinsic to callosal agenesis but also the frequently associated malformations of the limbic system.

Agenesis of Corpus Callosum↗

Isolation and characterization of Bsk, a growth factor receptor-like tyrosine kinase associated with the limbic system.

Neuronal degeneration has been shown to be involved in various neurological disorders. Growth/trophic factors and their receptors are known to be important for the regeneration and survival of neurons. We report here the molecular cloning of a receptor-like protein tyrosine kinase, bsk, (for brain specific kinase). Bsk is highly related to the eph/elk receptor-like kinase family members. Northern blot analysis shows that it is expressed specifically in the brain, with no expression detected in adult heart, spleen, lung, liver, skeletal muscle, and kidney. In situ hybridization analysis of adult mouse brain sections indicates that bsk is expressed at high levels in the hippocampus, tenia tecta, indusium griseum, and the piriform cortex, major components of the limbic system that are important for learning and memory. In addition, elevated levels of expression are found in other areas of the limbic system such as the amygdala, medial septum, and nucleus of the diagonal band, and in the olfactory bulb, which has close connections to the limbic system. The highest level of expression is found in the CA3 region of the hippocampus and the pyramidal cell layer of the piriform cortex. In 16.5 day mouse embryos, bsk is expressed predominantly in the primordial cortex of the telencephalon. An antibody against a C-terminal peptide of bsk recognized a 105 kD protein in the 16.5 day embryonic head extract. Our analysis shows that bsk is a growth factor receptor-like protein tyrosine kinase and that its greatest expression in the adult brain is associated with components of the limbic system.

Amino Acid Sequence↗

Stress-induced sensitization of the limbic system in ovariectomized rats is partly restored by cyclic 17beta-estradiol administration.

Chronic stress induces neurobiological alterations which have consequences for subsequent stress handling. In the current experiment, ovariectomized rats were subjected daily to a stressor for 21 days. Thereafter, the rats were treated for 21 days with 17beta-estradiol benzoate (10 microg/250 g, once every 4 days) or mirtazapine (10 mg/kg, daily). In this way, we were able to evaluate the ability of these compounds to reverse chronic stress-induced changes in the activity of the limbic system. After 21 days of recovery and treatment, the rats were re-exposed to the adverse environment of the initial stressor and perfused 2 h later. Ovariectomized rats displayed increased numbers of c-Fos-positive nuclei, after re-exposure to the stressor, in the paraventricular nucleus of the hypothalamus, dentate gyrus, medial prefrontal cortex and central and medial amygdala. Cyclic estradiol treatment attenuated the sensitization of the paraventricular nucleus and central amygdala. Mirtazapine increased the number of c-Fos-positive nuclei in the central amygdala and dentate gyrus. Long-term transcriptional changes induced by chronic stress were determined with DeltaFosB immunoreactivity. The medial prefrontal cortex showed an increased number of DeltaFosB-positive nuclei after chronic stress and this was not affected by estradiol or mirtazapine administration during recovery. In conclusion, cyclic estradiol administration reversed chronic stress-induced sensitization in the limbic system, the paraventricular nucleus and central amygdala of female rats, output regions of the limbic system involved in fear responses. Mirtazapine did not achieve this reversal of stress-induced aberrations in the limbic system after 21 days of treatment.

Amygdala↗

Regional cerebral blood volume (rCBV) and trasversal relaxation time (T2) mapping of the rat limbic system during pre-puberal and adult age.

We analyzed modifications in transversal relaxation time (T2) and regional cerebral blood volume (rCBV) in two areas of the limbic system, i.e., olfactory bulb (OB) and amygdala (AMY), in pre-puberty and post-puberty female rats. The aim of this work was to extend the knowledge about physiological modifications of these MRI parameters at different developmental phases. No significant difference was observed in T2 values of the OB between the two groups (pre-puberty: T2 = 86.92 +/- 8.57 ms, post-puberty: T2 = 88.11 +/- 13.06 ms; mean +/- S.D.). On the contrary T2 values of the AMY were significantly different (P = 0.0001) between the two groups (pre-puberty 76.08 +/- 3.2, post-puberty 81.77 +/- 11.77 ms). rCBV values of OB were significantly different (P = 0.0025) between pre-puberty (0.38 +/- 0.12 a.u.) and post-puberty female rats (0.15 +/- 0.09 a.u.). A significant decrease in rCBV (P = 5.1 x 10(-13)) between pre-puberty and post-puberty females (pre-puberty: 0.36 +/- 0.12, post-puberty: 0.07 +/- 0.05 a.u.) was also observed in the AMY. These findings suggest that in the limbic system, microvascular plasticity parallels neuronal maturation and indicate the importance of an appropriate baseline study in experiments dealing with the limbic system performed at different time-points.

Aging↗

Relationship between glutamate in the limbic system and hypothalamus-pituitary-adrenal axis after middle cerebral artery occlusion in rats.

OBJECTIVE: To investigate the features of glutamate activity in the limbic system and the effects of glutamate on the activation of the hypothalamus-pituitary-adrenal (HPA) axis throughout both acute cerebral ischemia and reperfusion. METHODS: The changes in glutamate content in the nervous cell gap, in corticotrophin releasing hormone (CHR) mRNA expression level in brain tissue, and in adrenocorticotropic hormone in blood plasma at different time-points after middle cerebral artery occlusion (MCAO) in rats were determined respectively with high-performance liquid chomatography (HPLC) and in situ hybridization. RESULTS: Glutamate content in the hippocampus and the hypothalamus increased rapidly at ischemia 15 minutes, and reached peak value (the averages were 21.05 mg/g +/- 2.88 mg/g and 14.20 mg/g +/- 2.58 mg/g, respectively) at 1 hour after middle cerebral artery occlusion. During recirculation, it returned rapidly to the baseline level. At 24 hours after reperfusion, it went up once more, and remained at a relative high level until 48 hours after reperfusion, and then declined gradually. CRH mRNA expression levels in the temporal cortex, hippocampus and hypothalamus were enhanced markedly at 1 hour ischemia and were maintained until 96 hours after reperfusion. At the same time, adrenocorticotropic hormone level in plasma was relatively increased. In the peak stage of reperfusion injury, there was a significantly positive correlation (n = 15, r = 0.566, P < 0.05) of the glutamate contents in the hypothalamus with the number of cells positive for CRH mRNA expression level in the hypothalamus. CONCLUSION: It is probable that the CRH system in the central nervous system is mainly distributed in the limbic system, and glutamate might be one of the trigger factors to induce excessive stress response in the HPA axis.

Animals↗

Establishment of status epilepticus by limbic system stimulation in previously unstimulated rats.

A syndrome of convulsive status epilepticus developed in 4 of 18 rats which had been treated with continuous sine wave stimulation incrementally raised to 40 microA through limbic system electrodes. The syndrome was characterized by recurrent behavioral seizures, continuous EEG spiking, and marked neuropathology. In three other animals, the stimulation treatment produced a syndrome of nonconvulsive status epilepticus manifested by an electrophysiologic record of continuous seizure activity, without accompanying tonic-clonic movements. The poststimulatory effect was correlated with the animals' response to the stimulation, and was independent of whether the electrode was positioned in the hippocampus or amygdala. If an animal developed repeated convulsive seizures during the stimulation, such seizures were likely to persist after the stimulation offset. These results indicate that persistent limbic system activation can produce a syndrome of recurrent seizures similar to that caused by either neurotoxic drugs or by limbic system activation in kindled rats.

Amygdala↗

Limbic system, the main focus of dementia syndrome--a study with MRI and PET.

Alzheimer disease and multi-infarct dementia are two entirely different diseases producing almost the same abnormalities as dementia syndrome. The statistical studies with MRI to locate the focus of dementia syndrome in the neocortex was an absolute failure. With MRI there is drastic atrophy and destruction of the amygdala and hippocampus suggesting the limbic system as the focus of dementia syndrome. Destruction of the limbic system in particular amygdala and hippampus produced the functional obstruction brought about by the marked reduction in the glucose utilization with PET in the bilateral temporal, parietal and occipital association cortices. Although this type constitutes only about 1/5 of all dementia patients. It is considered the fundamental type of dementia syndrome. Aside from this, there is a type wherein simultaneous and symmetrical reductions in glucose uterization of the frontal association cortex and the motor association cortex in the anterior part of the neocortex. This is referred to as type II. It constitutes about 4/5 of all dementia patients which is far more than type I. Based on these results, it is thought that limbic system is the main forcus of dementia syndrome.

Aged↗

Peripubertal ontogeny and estrogen stimulation of cholecystokinin and preproenkephalin mRNA in the rat hypothalamus and limbic system.

The neuropeptide cholecystokinin (CCK) is expressed in limbic system and hypothalamic nuclei that form a circuit that regulates the display of the female rodent reproductive behavior, lordosis. CCK mRNA and peptide levels fluctuate across the estrous cycle and have been shown to be modulated by estrogen exposure. The objective of these experiments was to examine the expression of CCK mRNA during postnatal development of this limbic-hypothalamic, lordosis regulating circuit, and to determine the age at which CCK mRNA expression becomes responsive to estrogen stimulation, by using quantitative in situ hybridization histochemistry. CCK mRNA levels were below the level of detectability within the circuit during the postnatal period, but increased during the peripubertal period. Rats were injected with either estradiol benzoate (EB), EB and progesterone, progesterone, or oil, and were killed 48 hours later on postnatal day (PND) 15, 20, and 25. Alternate brain sections were processed for CCK and preproenkephalin (PPE) mRNA in situ hybridization histochemistry. EB treatment induced CCK mRNA expression in the central portion of the medial preoptic nucleus and posterodorsal medial amygdala at PND 20 and 25, respectively. However, EB treatment increased PPE mRNA levels within the nuclei of the circuit at all ages examined. Progesterone had neither an independent nor additive effect on the EB induction of these neuropeptide messages. The estrogenic induction of CCK mRNA appears to be dependent on estrogen sensitive pathways of neurotransmission, or components of second messenger pathways which regulate CCK mRNA expression in the adult limbic-hypothalamic lordosis regulating circuit, which are not functional before PND 18-25.

Animals↗