A simple and fast densitometric method for the analysis of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area

Xavier, Léder Leal; Viola, Giordano Gübert; Ferraz, Anete Curte; Cunha, Cláudio Da; Deonizio, Janyana M. D.; Netto, Carlos Alexandre; Achaval, Matilde

doi:10.1016/j.brainresprot.2005.10.002

Cited by 157 publications

(96 citation statements)

References 17 publications

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“…The expression of CD8, S100 and Foxp3 was observed under a microscope (Carl zeiss, Oberkochen, Germany). According to the method developed by Xavie et al (13), we quantified the results of the immunostaining with the Imag Pro Plus 6.0 software (media Cybernetic, Silver Spring, mD, uSA). The measurement parameter was integrated optical density (IOD).…”

Section: Flow Cytometric Analysis For Cytotoxic T Lymphocytesmentioning

confidence: 99%

Modulated electro-hyperthermia enhances dendritic cell therapy through an abscopal effect in mice

et al. 2014

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Abstract. The aim of this study was to assess whether modulated electro-hyperthermia (mEHT) can induce an abscopal effect and thereby enhance the antitumor effects of immunotherapy. We used an intratumoral dendritic cell (DC) injection and mEHT to treat C3H/He mice inoculated with squamous cell carcinoma SCCVII cells in the left leg, and we assessed the whole body antitumor effects. Tumors were examined every two or three days in order to assess growth inhibition. The tumor-draining lymph nodes were removed to enable flow cytometric analysis of CD3 + and CD8 + cells, whereas immunohistochemistry was used to assess CD8, S100 and Foxp3 expression in the tumors. Additionally, GP96 expression in the tumors from the different treatment groups was measured. In the control group, the mean tumor volume was larger than that in other groups. These results indicated that the combination therapy of an intratumoral DC injection and mEHT evoked systemic antitumor activity. A larger number of CD3 + and CD8 + cells were detected by flow cytometric analysis in the DC plus mEHT treatment group. Tumor tissue immunostaining showed that CD8 and S100 were more strongly expressed in the DC plus mEHT treatment group, although Foxp3 expression was much higher in the control group. The GP96 gene expression level in the mEHT group was significantly different from the expression level in the control group. An abscopal effect may be induced by mEHT, and the effect of immunotherapy with DCs was strongly enhanced by the overexpression of GP96. GP96 is thought to be one of the molecules explaining the abscopal effect. Direct intratumoral administration of DCs and mEHT may be a feasible future treatment strategy.

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Section: Flow Cytometric Analysis For Cytotoxic T Lymphocytesmentioning

confidence: 99%

Modulated electro-hyperthermia enhances dendritic cell therapy through an abscopal effect in mice

et al. 2014

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“…Cell loss in the STN and the correct location of NMDA injection were determined using two-dimensional images of Nissl-stained sections (28,29).…”

Section: Histological Evaluation Of Stn Lesionmentioning

confidence: 99%

“…Rats were deeply anesthetized with 200 mg/kg sodium thiopental, ip, and transcardially perfused with saline followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB), pH 7.4 (24,25,28). After perfusion, the brains were removed from the skulls, postfixed in the same solution at room temperature for 2 h and cryoprotected by immersion in 30% sucrose solution in PB at 4°C until they sank.…”

Section: Th Immunohistochemistrymentioning

confidence: 99%

“…The brains of all animals were fixed and postfixed for the same time in the same batch of solution rigorously processed at the same time, and the sections were incubated in an identical medium for the same period of time. This precaution was taken to avoid overreaction, differences in chromogen reaction, saturation of absorbance, and changes in background levels (24,25,28,30). Coronal sections of the SNpc were selected according to the atlas of Paxinos and Watson (26) and the readings were obtained between the following coordinates: interaural 4.2 mm, bregma -4.8 mm and interaural -2.7 mm, bregma -6.3 mm.…”

Section: Th Immunohistochemistrymentioning

confidence: 99%

“…To determine neuronal density, the sum of TH-ir neurons located inside this square or intersected by the lower and/ or right edge of the square were counted. The neurons that were intersected by the upper and/or left edge of the square were not counted (24,25,28).…”

Section: Th Immunohistochemistrymentioning

confidence: 99%

See 2 more Smart Citations

Lesion of the subthalamic nucleus reverses motor deficits but not death of nigrostriatal dopaminergic neurons in a rat 6-hydroxydopamine-lesion model of Parkinson's disease

Rizelio

Szawka

Xavier

et al. 2010

Braz J Med Biol Res

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Cellular components of the human medial amygdaloid nucleus

Dall'Oglio

Xavier

Hilbig³

et al. 2012

J of Comparative Neurology

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The medial nucleus (Me) is a superficial component of the amygdaloid complex. Here we assessed the density and morphology of the neurons and glial cells, the glial fibrillary acidic protein (GFAP) immunoreactivity, and the ultrastructure of the synaptic sites in the human Me. The optical fractionator method was applied. The Me presented an estimated mean neuronal density of 1.53 × 10⁵ neurons/mm³ (greater in the left hemisphere), more glia (72% of all cells) than neurons, and a nonneuronal:neuronal ratio of 2.7. Golgi-impregnated neurons had round or ovoid, fusiform, angular, and polygonal cell bodies (10-30 μm in diameter). The length of the dendrites varied, and pleomorphic spines were found in sparsely spiny or densely spiny cells (1.5-5.2 spines/dendritic μm). The axons in the Me neuropil were fine or coarsely beaded, and fibers showed simple or notably complex collateral terminations. The protoplasmic astrocytes were either isolated or formed small clusters and showed GFAP-immunoreactive cell bodies and multiple branches. Furthermore, we identified both asymmetrical (with various small, clear, round, electron-lucent vesicles and, occasionally, large, dense-core vesicles) and symmetrical (with small, flattened vesicles) axodendritic contacts, also including multisynaptic spines. The astrocytes surround and may compose tripartite or tetrapartite synapses, the latter including the extracellular matrix between the pre- and the postsynaptic elements. Interestingly, the terminal axons exhibited a glomerular-like structure with various asymmetrical contacts. These new morphological data on the cellular population and synaptic complexity of the human Me can contribute to our knowledge of its role in health and pathological conditions.

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A simple and fast densitometric method for the analysis of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area

Cited by 157 publications

References 17 publications

Modulated electro-hyperthermia enhances dendritic cell therapy through an abscopal effect in mice

Modulated electro-hyperthermia enhances dendritic cell therapy through an abscopal effect in mice

Lesion of the subthalamic nucleus reverses motor deficits but not death of nigrostriatal dopaminergic neurons in a rat 6-hydroxydopamine-lesion model of Parkinson's disease

Cellular components of the human medial amygdaloid nucleus

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