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Objective To study the effect of ethonal on the dopaminergic system by analyzing the altered expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the brain of ethanol-treated rats. Methods Sixty Wistar rats were selected and divided into control group and ethanol-treated group, 30 per group, the ethanol-treated rats were treated with 20% ethanol for 6 months. Immunohistochemistry, flow cytometry and Western blotting were used to analyze the altered expression of TH and DAT in the DA energic system in different brain regions of the ethanol treated rats. Results 1. Immunohistochemistry showed the mean gray value of TH in substantia nigra(SN)-ventrotegmental area (VTA), caudae putamen (Cpu) and nucleus accumbens (NACC), DAT in Cpu and NACC of the ethanol were smaller than those in control (P<0.05). 2. Flow cytometry showed the expression of TH in middle brain of the ethanol-treated rats increased significantly compared with the control(P<0.05). 3. Western blotting showed the ratio of IA of TH/β-actin and DAT/β-actin in different brain regions of the ethanol-treated rats were larger than those in control(P<0.05).Conclusion Ethanol treatment increases the expression of TH and DAT in rat brain.
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@#ObjectiveTo explore the effects of the conjugate prepared from the cholera toxin B subunit(CB) and nerve growth factor(NGF) on the spatial learning and memory abilities and cholinergic function.MethodsThe conjugate of CB-NGF was prepared by the improved sodium metaperiodate method and nasally administrated to the β-amyloid protein(Aβ25-35) induced amnesic mice for 7 days with 2 dosage (7-5 μg/d、15 μg/d). Spatial learning and memory abilities were evaluated by Morris water maze and cholinergic function was assessed with the choline acetyl transferase (ChAT) immunohistochemical methods.ResultsMorris water maze test showed that the escape latency in Aβ25-35-treated mice prolonged and the staying time reduced in the crossed first quadrant where the platform had been located, compared with the control mice (P<0-01). In addition, the number of ChAT positive neuron declined in the model mice(P<0-001). CB-NGF nasal administration significantly shortened the escape latency and elevated the staying time and number of ChAT positive neuron(P<0-01).ConclusionCB-NGF treatment can improve the spatial and memory performance which may involve the neuroprotection to cholinergic system.
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Objective To study serotonin transporters(SERT) in postmortem human brains of Parkinson's disease(PD).Methods Immunoautoradiography was used to show SERT distribution in postmortem PD brains.ResultsIn comparison with healthy group,SERT decreased in dorsal raphe nucleus,substantia nigra and striatum in PD brains.Quantitative analysis showed that 25.9%、31.1%、27.2% and 24.7% of labeling intensity of SERT decreased in putamen,caudate nucleus,substantia nigra and ventrotegmental area as compared with corresponding control respectively.Among the four regions studied of dorsal raphe nucleus,SERT labeling intensity significantly decreased by 38.9%,37.3%,16.5% and 37.0% of corresponding control respectively in ventral part,dorsal part,caudal part and interfascicular part.Median raphe nucleus did not show the decreased SERT labeling.ConclusionDecreased SERT expression in three regions of postmortem PD brains indicates that a dysfunction of serotonergic raphe system may contribute to the etiology of Parkinson's disease.
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Objective To study the altered expression of dopamine transporter (DAT) in substantia nigra and striatum in postmortem human brain of Parkinson’s disease (PD). Methods Immunoautoradiography was used to reveal DAT distribution in postmortem human brain. Results Strongly labeling signal of DAT was mainly found in the substantia nigra, the putamen and the caudate nucleus in controls. In contrast, it was drastically reduced in the putmen and the dorsolateral caudate nuclus in PD brains, but the ventromedial part of the caudate nucleus showed a significant sparing adjacent to the border of the lateral ventricle. In the substantia nigra, the ventral and the lateral parts of the substantia nigra showed an obvious decreasing of DAT and the reducing degree of DAT labeling signals in those regions is smaller than that in the putamen and the caudate nucleus. Quantitative analysis revealed that 90.9% and 66.7% of the labeling intensity of DAT were decreased in the putamen and the caudate nucleus as comparing with the corresponding controls respectively (P
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Objective Immunoreactive intensity of dopamine transporter(DAT)was quantitatively analyzed in the different regions of postmortem human brain in order to provide evidence in selecting an appropriate reference region for neuroimaging in measurement of the altered DAT.Methods The brain tissue blocks taken from 8 male cadavers were paraffin-embedded and sectioned,and immunoautoradiography was used to display the difference of DAT immunoreactive intensity in the substantia nigra,putamen and caudate nucleus,cingulate cortex,frontal cortex,occipital cortex and cerebellar cortex.Results The highest DAT density was found mainly in the substantia nigra,caudate nucleus and putamen,while the lowest density was only seen in cerebellar cortex.Quantitative analysis revealed that the intensity of DAT immunoreactivity(DAT-IR)in cerebellar cortex was respectively 1/3.50,1/3.72,1/1.28 of that in frontal cortex,cingulate cortex and occipital cortex,and it was only 1/8.33,1/11.67,1/8.56 that of substantia nigra,caudate nucleus and putamen.Conclusions The cerebellar cortex has the lowest DAT as compared to other examined brain regions,and it can be used for a reference region in neuroimaging to detect altered DAT.
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Objective To explore the effect of androgen on learning and memory ability and neurons in hippocampal CA1 region in senescence accelerated mouse prone strain/8(SAMP8).Methods Thirty 7-month-old male SAMP8 were randomly divided into sham-operation control group,castrated group and androgen replacement therapy after castration group.The dose of testosterone undecanoate(TU) was 37.4mg/(kg?15d).The capability of learning and memory was observed 45 days later through the Morris water maze(MWM) test and the change of neurons in hippocampal CA1 region was detected and analyzed by HE staining,immunohistochemal method and computer pathological image analysis system.Results 1.In the MWM test,the escape latency of castrated group were significantly prolonged(P0.05).2.With HE staining,neurons in hippocampal CA1 region of castrated group were found with diffused vacuolar degeneration,and sparse and disordered cellular arranpement.The cell nucleuses were karyochrome and karyopycnosis.The number and optical density of A? immune positive neurons were markedly higher than those of other groups(P