It is well known that the status epilepticus induced by the administration of pilocarpine to lithium-pretreated rats is a role model to study for the cholinergic system in epileptogenesis and the pathogenesis of status epilepticus. Although the researches for the expression of the c-fos proto-oncogene in seizure models have been studied, the exact role of the c-fos expression is still uncertain. To evaluate the characteristics of lithium-pilocarpine seizure model, we designed this study by following three steps; (1) the analysis of clinical manifestations by video, and electroencephalogram through implanted cortical electrodes during the course of status epilepticus induced by intraperitoneal administration of lithium chloride (5 mEq/kg) followed by pilocarpine(50 mg/kg) in Sprague-Dawley rats, (2) Fos expression and the time course of Fos appearance by the immunocytochemistry, (3) Neuropathologic change by cresyl violet stain. The presentation of clinical manifestations were cholinergic symptoms and signs, stereotyped behaviour, motor seizures, and status epilepticus in order. Electroencephalographic findings showed five patterns : (I) discrete seizure with interictal slowing; (ii) merging seizures with waxing and waning ictal discharges; (iii) continuous ictal discharges; (iv) continuous ictal discharges with flat periods; and (v) periodic epileptiform discharges on a flat background. The neuroanatomical sites of Fos expression were the taenia recta, anterior olfactory nucleus, olfactory tubercle, piriform cortex, entorhinal cortex, amygdala, septum, accumbens, caudate-putamen, hippocampus, dentate gyrus, thalamus, and cerebral neocortex. The Fos immunostaining appeared first in the taenia tecta, anterior olfactory nucleus, olfactory tubercle, and piriform cortex at 1 hour after pilocarpine injection, and was maximal in the most areas of cerebral cortex and limbic area between 4 and S hours. The thalamus and the caudate-putamen became stained after 4 hours. In the hippocampal formation, firstly, the Fos was stained maximally in the dentate gyrus at 3 hours followed by in the CA1, CA2, and CA3 regions. The Fos was disappeared in the dentate gyrus and CA2 region of hippocampus within 18 hours, but became stained sustainly in the CA3 and CA1 regions of hippocampus at 24 hours. Llght microscopic findings revealed widespread brain damage. The neuropathological changes were found within the anterior olfactory nucleus, piriform cortex, entorhinal cortex, thalamus, hippocampal formation, amygdaloid complex, lateral septum, neocortex and substantia nigra. There were only swollen and edematous change of neurons at 1 hour, but severely shrunken and darkened neuronal degeneration and neuronal loss at 72 hours. The neuronal degeneration and loss in hippocampal formation appeared severe in the CA1 and hilum, moderate in CA2, and mild in CA3 and dentate gyrus. In conclusion, it was suggested that cholinergic system (muscarinic receptor) played an important role in the induction of the seizure because Fos was expressed in the brain areas containing muscarinc receptor and the lithium-pilocarpine seizure was a good model to study for the status epilepticus.
Amygdala ; Animals ; Brain ; Cerebral Cortex ; Dentate Gyrus ; Electrodes ; Electroencephalography* ; Entorhinal Cortex ; Genes, fos ; Hippocampus ; Immunohistochemistry ; Lithium Chloride ; Neocortex ; Neurons ; Olfactory Pathways ; Pilocarpine ; Rats ; Rats, Sprague-Dawley ; Seizures* ; Status Epilepticus ; Substantia Nigra ; Taenia ; Thalamus ; Viola

Intranasal administration provides a method of bypassing the blood brain barrier, which separates the systemic circulating system and central interstitial fluid, and directly delivering drugs to the central nervous system. This method also circumvents first-pass elimination by the liver and gastrointestinal tract. In the present study, the authors investigated intranasal siRNA delivery efficiency by using FITC-labeled transfection control siRNA and a genespecific siRNA. The localization of fluorescence-tagged siRNA revealed that siRNA was delivered to cells in the olfactory bulb and that the level of the siRNA target gene (alpha B-crystallin) was significantly reduced in the same area. siRNA was delivered to processes as well as nuclei and cytoplasm. At 12 hrs after intranasal delivery, siRNA-mediated target gene reduction was observed in other more distally located brain regions, for example, in the amygdala, entorhinal cortex, and hypothalamus. Target gene knockdown was demonstrated by double immunohistochemistry, which demonstrated alpha B crystallin expression depletion in more than 70% of cells at 12 hrs after the intranasal delivery. siRNA-mediated target gene suppression was detected not only in neurons but in glia, for example, astrocytes. These results indicate that intranasal siRNA delivery offers an efficient means of reducing specific target genes in certain regions of the brain and of performing gene knockdown-mediated therapy.
Administration, Intranasal ; alpha-Crystallin B Chain ; Amygdala ; Astrocytes ; Blood-Brain Barrier ; Brain ; Central Nervous System ; Cytoplasm ; Entorhinal Cortex ; Extracellular Fluid ; Gastrointestinal Tract ; Gene Knockdown Techniques ; Hypothalamus ; Immunohistochemistry ; Liver ; Neuroglia ; Neurons ; Olfactory Bulb ; RNA, Small Interfering ; Transfection

OBJECTIVE: To explore the olfactory abilities in patients with allergic rhinitis (AR), analyze the correlation between olfactory bulb (OB) volume with depth of olfactory sulcus (OS) and olfactory function in patients with AR. METHOD: One hundred patients with AR were compared with one hundred controls in terms of olfactory function T&T testing, OB volume and depth of OS assessed with magnetic resonance imaging (MRI). T&T testing and MRI were done after a year in 100 AR patients,the results were compared with the initial results. RESULT: The OB volume in AR patients was (29.53±3.95) mm3 on the left, (29.67±14.21)mm3 on the right, (29.61±4.05) mm3 on average; The OB volume in controls was (48.93±6.73)mm3 on the left side, (48.81±7.43)mm3 on the right side, (48.85±7.11)mm3 on average; The OB volume in AR patients was less then the control group(t= 6.321, 6.141, 6.221, P<0.01). The OS depth had no statistical difference between AR patients and controls (t=1.032, 0.972, 0.991, P>0.05). Olfactory discriminate threshold was negatively correlated with OB volume in AR patients (r=-0.46, P<0.05); and it was no correlated with depth of OS (r=-0.012,P>0.05). Among 100 followed-up AR patients, 43 showed increased in OB volume and olfactory function after a year, but there was no statistical difference (t= 0. 811,0. 843, 0.826, P>0.05; Z=1.911, P>0.05) ,and the other 57 showed no significant changes of OB volume and olfactory function. CONCLUSION In AR patients, the OB volume and olfactory function decreased, but the depth of OS had no significant changes. The OB volume is correlated with olfactory function, while the depth of OS is no correlated with olfactory function. Conservative treatment had some clinical significance on the recovery of olfactory function in patients with AR.
Humans ; Magnetic Resonance Imaging ; Olfaction Disorders ; etiology ; Olfactory Bulb ; pathology ; physiopathology ; Prefrontal Cortex ; Rhinitis, Allergic ; complications ; Smell

We investigated a topographical distribution of managanese, and immunohistochemical density of tyrosine hydroxylase (TH), and histopathologic findings in globus pallidus and substantia nigra according to manganese dose and time course in the brain of rats which received MnCl2 intravenously. Topographical distribution of manganese was also investigated after injection of FeCl2. The manganese concentrations of brain in control and experimental group were highest in pituitary gland and thalamus, and lowest in the cerebral cortex. The manganese concentration of blood was increased proportionally to the dose administered, and the biological half-life of blood manganese was between 21 and 42 days. The manganese concentrations of brain were increased proportionally to the dose, and increase rate was highest in olfactory bulb, and the biological half-lives of brain manganese ranged from 42 days to 90 or more days; the longest were observed in pituitary gland, medulla oblongata and cerebral cortex. In case of administration of FeCl2, the manganese concentrations of brain were higher than that of control group in dose of 2.5 mg/kg, and decreased proportionally to the administered dose, resulting in lower level compared with control group in high dose of FeCl2 administered. Significantly decreased number of nerve cell and increased gliosis in globus pallidus were observed in experimental group, which were closely correlated with the duration after manganese injection, but no significant change of number of nerve cell expressing TH and gliosis were observed in substantia nigra. Density of immunohistochemical reaction for TH in globus pallidus made little difference between control and experimental group. These results suggest that pathology of manganese intoxication is caused by the loss of nerve cells in globus pallidus, and closely correlated with the duration after manganese exposure.
Animals ; Brain* ; Cerebral Cortex ; Gliosis ; Globus Pallidus ; Half-Life ; Manganese* ; Medulla Oblongata ; Neurons ; Olfactory Bulb ; Pathology ; Pituitary Gland ; Rats* ; Substantia Nigra ; Thalamus ; Tyrosine 3-Monooxygenase

OBJECTIVES: Peripheral benzodiazepine receptor has been suggested to be associated with the relief of anxiety response induced by stresses. This study was designed to observe the anxiolytic activity of peripheral benzodiazepine receptor. METHODS: Male Sprague-Dawley rats, weighing 200-250g were forced to suffer an immobilization stress for 2 hours. The level of anxiety by immobilization was performed by an elevated plus maze and was evaluated by the number of [3H]Ro5-4864 binding sites in the olfactory bulb. RESULTS: Saturation experiments followed by scatchard anlayses of the results showed that the density of peripheral benzodiazepine receptor increased and the affinity of the peripheral benzodiazepine receptor remained unchanged. It was found that there was no significant change in the cerebral cortex. Pretreatment with clonazepam, a central benzodiazepine receptor agonist, before an immobilization stress abolished the anxoius response on the performance of plus maze. In this group, upregulation of peripheral benzodiazepine receptor of olfactory bulb was not observed. Ro5-4864, a peripheral benzodiazepine receptor agonist, elicited an increase of anxiolytic response on the performance of plus maze. Progesterone, a precursor of neuroactive steroid, also increased anxiolytic response on the performance of plus maze. Pretreatment with PK11195, a peripheral benzodiazepine receptor antagonist, abolshed the anxiolytic effect of progesterone. CONCLUSIONS: From these results, it could be concluded that peripheral benzodiazepine receptor is closely associated with the relief of acute stress induced anxiety response via an increase of synthesis of neuroactive steroid.
Animals ; Anti-Anxiety Agents ; Anxiety* ; Benzodiazepines* ; Binding Sites ; Cerebral Cortex ; Clonazepam ; Humans ; Immobilization ; Male ; Olfactory Bulb ; Progesterone ; Rats* ; Rats, Sprague-Dawley ; Receptors, GABA-A* ; Up-Regulation

The frontal cortex of rat is innervated by dopaminergic pathway(mesocortical pathway) arising from ventral tegmental area. Several studies have suggested that mesocortical dopaminergic neurons may modulate the function of dopaminergic neurons at subcortical sites. The effect of lesions of the dopaminergic nerve terminals in the medial prefrontal cortex of the rat on dopamine D1 and D2 receptors within the striatum and olfactory tubercle has been investigated. Bilateral 6-hydroxy-dopamine lesions were stereotaxically placed in the medial prefrontal cortex. Animal were pretreated with desipramine to block the uptake of neurotoxin into noradrenergic terminals and to make it more selective for dopamine terminal. After 2weeks later, we examined the changes of D1 and D2 receptors in caudate-putamen and nucleus accumbens by quantitative autoradiography using the specific D1 antagonist [3H]SCH23390 and D2 antagonist [3H]spiperone. The results shows that D1 receptor at striatum was up regulated 2weeks after destruction of dopamine terminals within medial prefrontal vortex of the rat. This findings suggest that frontal cortical dopamine system may regulate the dopamine system in corpus striatum.
Animals ; Autoradiography ; Corpus Striatum ; Desipramine ; Dopamine* ; Dopaminergic Neurons ; Nucleus Accumbens ; Olfactory Pathways ; Oxidopamine* ; Prefrontal Cortex* ; Rats* ; Receptors, Dopamine* ; Ventral Tegmental Area

As it has been reported that the depolarization induced acetylcholine (ACh) release is modulated by activation of presynaptic A-1 adenosine heteroreceptor and various evidence suggest that indicate the A-2 adenosine receptor is present in the striatum, this study was undertaken to delineate the role of adenosine receptors on the striatal ACh release. Slices from the rat striatum were equilibrated with (3H)choline and then the release amount of the labelled product, (3H)ACh, which was evoked by electrical stimulation (rectangular pulses, 3 Hz, 2 ms, 24 mA, 5 Vcm-1, 2 min), was measured, and the influence of various agents on the evoked tritium outflow was investigated. And also, quantitative receptor autoradiography and drug-receptor binding assay were performed in order to confirm the presence and characteristics of A-1 and A-2 adenosine receptors in the rat striatum. Adenosine (10 ~ 100 micrometer) and N-6-cyclopentyladenosine (CPA, 1 ~ 100 micrometer) decreased the (3H)ACh release in a dose-dependent manner without changing the basal rate of release in the rat striatum. The reducing effects of ACh release by adenosine and CPA were abolished by 8-cyclopentyl-1,3-dipropy-lxanthine (DPCPX, 2 micrometer), a selective A-1 adenosine receptor antagonist, treatment. The effect of adenosine was potentiated markedly by 3,7-dimethyl-1-propargylxanthine (DMPX, 10 micrometer), a specific A-2 adenosine receptor antagonist. 2-P-(2-carboxyethyl)phenethylamimo-5'-N- ethylcarboxamidoadenosine hydrochloride (CGS-21680C), in concentrations ranging from 0.01 to 10 micrometer, a recently introduced potent A-2 adenosine receptor agonist, increased the(3 H)ACh release in a dose related fashion without changing the basal rate of release. These effects were completely abolished by DMPX (10 micrometer). In autoradiogaphy experiments, (3H)2-chloro-N-6-cyclopentyladenosine ((3 H)CCPA) bindings were highly localized in the hippocampus and the cerebral cortex. Additionally, lower levels of binding were found in the striatum. However, (3H)CGS-21680C bindings were highly localized in the striatal region with the greatest density of binding found in the caudate nucleus and putamen. Lower levels of binding were also found in the nucleus accumbens and olfactory tubercle. In drug-receptor binding assay, binding of (3H)CCPA to A-1 adenosine receptors of rat striatal membranes was inhibited by CPA (K-i = 1.6nM) and N-ethylcarboxamidoadenosine (NECA, K-i = 12.9 nM), but not by CGS-21680C (K-i = 2609.2 nM) and DMPX (K-i = 19,386 nM). In contrast, (3H)CGS-21680C binding to A-2 adenosine receptors was inhibited by CGS-21680C (K-i = 47.6 rim) and NECA (K-i = 44.9 nM), but not by CPA (K-i = 2099.2 nM) and DPCPX (K-i = 19,207 nM). The results presented here suggest that both types of A-1 and A-2 adenosine heteroreceptors exist and play an important role in ACh release in the rat striatal cholinergic neurons.
Acetylcholine* ; Adenosine* ; Adenosine-5'-(N-ethylcarboxamide) ; Animals ; Autoradiography ; Caudate Nucleus ; Cerebral Cortex ; Cholinergic Neurons ; Electric Stimulation ; Hippocampus ; Membranes ; Nucleus Accumbens ; Olfactory Pathways ; Putamen ; Rats* ; Receptors, Purinergic P1* ; Tritium

The Kallmanns syndrome is the most common form of isolated hypogonadotropic hypogonadism in which anosmia or hyposmia resulting from agenesis of hypoplasia of the olfactory lobes is associated with LHRH deficiency, This syndrome is genetically heterogeneous and can be trans-mitted as an X-linked, autosomal dominant or autosomal recessive trait. The hypogonadotropic hypogonadism results in absent or incomplete pubertal development and may be associated with anosmia or hyposmia, mid-line defect(color blindness, cleft-lip or
Blindness ; Cryptorchidism ; Epiphyses ; Femur Neck ; Gonadotropin-Releasing Hormone ; Growth Plate ; Head ; Humans ; Hypogonadism ; Kallmann Syndrome ; Male ; Olfaction Disorders ; Olfactory Cortex ; Slipped Capital Femoral Epiphyses

Using in situ hybridization technique with digoxigenin-labelled riboprobe, study on the expression of hsp 70 mRNA in the developing mouse brain was performed. The results obtained are as follows; 1. In embryonic day 16 group, cells with strong reactivity to hsp70 mRNA were found in spinal cord. In neuroepithelial layer lining fourth ventricle and external granular layer of cerebellum, moderate reactivity was observed. But the reactivity was weak in the forebrain including cerebral cortex, diencephalon and olfactory bulb. 2. In embryonic day 18 group, the regional pattern of hsp70 mRNA expression was similar to that of embryonic day 16 group. In medulla oblongata, however, stronger reactivity was found in the embryonic day 18 group. 3. In postnatal day 0 mice group, cells with moderate or strong reactivity to hsp70 mRNA were found in the overall area of central nervous system, Especially, cells with moderate reactivity were found in the dentate gyrus of hippocampus, and the supragranular cortical plate and subplate neocortex. 4. In postnatal day 2 mice group, cells with moderate or strong reactivity to hsp70 mRNA were found in the same pattern as in postnatal day 0 group. Further differentiation of cerebral cortex and cerebellum was found. 5. Strong expression of hsp70 mRNA was found in the areas with high rate of cell division. In general, the area of expression moved to more rostral area in central nervous system as development proceeds. Above results suggest that hsp70 play an important role in the development and differentiation of central nervous system.
Animals ; Brain* ; Cell Division ; Central Nervous System ; Cerebellum ; Cerebral Cortex ; Dentate Gyrus ; Diencephalon ; Fourth Ventricle ; Heat-Shock Proteins* ; Hippocampus ; Hot Temperature* ; HSP70 Heat-Shock Proteins* ; In Situ Hybridization ; Medulla Oblongata ; Mice* ; Neocortex ; Olfactory Bulb ; Prosencephalon ; RNA, Messenger ; Spinal Cord

Voltage dependent calcium channels mediate wide variety of physiological functions including neurotransmitter release, neurite outgrowth, and gene expression in neurons. omega-Conotoxin-sensitive N-type calcium channels are exclusively expressed in nervous system and involved in the control of neurotransmitter release from neurons. In this experiment, I have investigated human chromosomal location and rat neuronal distribution of N-type voltage dependent calcium channel alpha1, subunit [alpha1B]. I have localized human alpha1B subunit gene to the long arm of chromosome 9[9q34] by fluorescent in situ hybridization. The distribution of rat alphaB1 subunit mRNA has been examined in the rat brain by in situ hybridization histochemistry and high level of alpha1B subunit mRNA has been observed in olfactory bulb, anterior olfactory nucleus, cerebral cortex, piriform cortex, hippocampus, dentate gyrus, parabrachial nucleus, and cerebellum and low level of expression was also found in other areas of rat brain.
Animals ; Arm ; Brain* ; Calcium Channels* ; Calcium Channels, N-Type ; Calcium* ; Cerebellum ; Cerebral Cortex ; Dentate Gyrus ; Gene Expression ; Hippocampus ; Humans ; In Situ Hybridization ; In Situ Hybridization, Fluorescence ; Nervous System ; Neurites ; Neurons ; Neurotransmitter Agents ; Olfactory Bulb ; Rats ; RNA, Messenger