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Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/S1385-299X(04)00031-5

引用次数: 0

Nitric oxide synthase in rat visual cortex: an immunohistochemical study 大鼠视觉皮层一氧化氮合酶的免疫组织化学研究

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2004.01.004

Tagrid Yousef , Ute Neubacher , Ulf T Eysel , Maxim Volgushev

{"title":"Nitric oxide synthase in rat visual cortex: an immunohistochemical study","authors":"Tagrid Yousef , Ute Neubacher , Ulf T Eysel , Maxim Volgushev","doi":"10.1016/j.brainresprot.2004.01.004","DOIUrl":"10.1016/j.brainresprot.2004.01.004","url":null,"abstract":"<div><p><span>The aim of the present study was to identify the distribution of two isoforms of the nitric oxide synthase (NOS), the neuronal (nNOS) and the endothelial (eNOS) form, in rat </span>visual cortex. Immunohistochemical localisation of each NOS isoform was studied with three tissue-processing protocols. In the first one, immunohistochemical reactions were made on 30-μm-thick sections with membrane detergents, Triton or Saponin, used to increase the permeability of the tissue for the antibodies. In the second protocol, we excluded these detergents from all solutions to avoid a destruction of the cellular membrane. In the third protocol, we used thin paraffin sections (5 μm thick) to assure delivery of the antibodies to intracellular structures. Our data demonstrate, that both neuronal and endothelial isoforms of the NOS are present in the visual cortex. Among the neurones labelled by the antibodies against eNOS or nNOS, some excitatory cells were definitely present. nNOS immunopositive were neurones and a dense network of fibres, presumably axons. Some of the neurones were heavily labelled in a Golgi-like manner, while others showed only weak labelling. eNOS immunopositivity was found in the blood vessels and in neurones. eNOS positive neurones were much more numerous than nNOS-containing cells, and represent about 60% of the cortical cells. However, with antibodies against eNOS, we never observed neurone-specific cell features. The NOS-containing cells found in our present study represent a possible morphological substrate for production of nitric oxide (NO).</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 57-67"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2004.01.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 22

In vitro model of the outer blood–retina barrier 体外血视网膜屏障模型的建立

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2003.12.002

H Steuer, A Jaworski, D Stoll, B Schlosshauer

{"title":"In vitro model of the outer blood–retina barrier","authors":"H Steuer, A Jaworski, D Stoll, B Schlosshauer","doi":"10.1016/j.brainresprot.2003.12.002","DOIUrl":"10.1016/j.brainresprot.2003.12.002","url":null,"abstract":"<div><p><span>The outer blood–retina barrier (BRB) is formed by the retinal pigment epithelium<span> (rpe) and functions similarly to the blood–brain barrier (BBB). In contrast to the BBB, which is composed of a myriad of capillaries, the rpe can in principle be prepared as an intact planar tissue sheet without disruption of its barrier and carrier functions. Both a rapid and gentle procedure to isolate porcine rpe and a method to implement the harvested rpe in drug penetration testing are presented. Enucleated eyes were flat-mounted and the RPE/choroid tissue sheets with or without the retina were isolated. Fluorescence microscopy based on double-labeling with propidium iodide/calcein and scanning electron microscopy revealed well-preserved cell and tissue architecture. For drug evaluation, specimens were immobilized as the interface between test compartments in a dual-chamber device. Ten different test agents were added to one chamber at defined concentrations. After an incubation time of 30 min at 37 °C permeated drug levels in both compartments were quantified by HPLC-tandem mass spectrometry or HPLC with fluorescence detection. Sodium </span></span>fluorescein used as a barrier marker indicated that the rpe model had excellent seal integrity. The use of a representative subset of pharmaceuticals with known BBB permeability characteristics demonstrated that the rpe model had a large permeability dynamic range (factor >350). These findings showed that the model represents a valuable tool for the investigation of the blood barrier penetration of test compounds.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 26-36"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.12.002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 59

Two immunocytochemical protocols for immunofluorescent detection of c-Fos positive neurons in the rat brain 大鼠脑c-Fos阳性神经元免疫荧光检测的两种免疫细胞化学方案

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2004.01.003

Peter Kobelt , Johannes J Tebbe , Ines Tjandra , Hi-Gung Bae , Jens Rüter , Burghard F Klapp , Bertram Wiedenmann , Hubert Mönnikes

{"title":"Two immunocytochemical protocols for immunofluorescent detection of c-Fos positive neurons in the rat brain","authors":"Peter Kobelt , Johannes J Tebbe , Ines Tjandra , Hi-Gung Bae , Jens Rüter , Burghard F Klapp , Bertram Wiedenmann , Hubert Mönnikes","doi":"10.1016/j.brainresprot.2004.01.003","DOIUrl":"10.1016/j.brainresprot.2004.01.003","url":null,"abstract":"<div><p>The immediate-early-gene product c-Fos is a well known marker of neuronal activation in the central nervous system. Thus, immunocytochemical methods to detect c-Fos in the brain are important tools in experimental studies that aim to map activated brain areas on a cellular level. Accordingly, we describe here two alternative protocols for c-Fos detection which are based on an indirect immunofluorescence technique. In fact, both methods allow an excellent and specific visualisation of c-Fos immunoreactive neurons in brain areas, e.g. the hypothalamus. The first protocol is more economical and faster in its execution and useful for observing brain sections using a confocal laser scanning microscope with the intention to perform doublestaining, since in all optical magnification steps (10×–63×) only a low unspecific background staining is visible. Furthermore, this method yields even fluorescent signals which are not detectable with a conventional fluorescence-microscope at lower magnification (10×). The second protocol contains an additional signal amplification step and allows signal detection also with a conventional fluorescence-microscope at lower magnification (10×); it is useful for rapid quantification of c-Fos immunoreactive neurons in the rat brain, but because of moderate unspecific background staining at higher magnification it is less suitable for doublestaining.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 45-52"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2004.01.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 34

Free colour illustrations in the online version of articles 免费彩色插图在文章的在线版本

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/S1385-299X(04)00030-3

引用次数: 0

Contents of Brain Research Protocols Volume 12 脑研究协议第12卷的内容

Brain research. Brain research protocols Pub Date : 2004-02-01 DOI: 10.1016/S1385-299X(04)00010-8

引用次数: 0

Volumetric evaluation of the ischemic lesion size with serial MRI in a transient MCAO model of the rat: comparison of DWI and T1WI 瞬态MCAO大鼠模型的连续MRI体积评价:DWI与T1WI的比较

Brain research. Brain research protocols Pub Date : 2004-02-01 DOI: 10.1016/j.brainresprot.2003.11.004

Simon Nagel , Simone Wagner , James Koziol , Britta Kluge , Sabine Heiland

{"title":"Volumetric evaluation of the ischemic lesion size with serial MRI in a transient MCAO model of the rat: comparison of DWI and T1WI","authors":"Simon Nagel , Simone Wagner , James Koziol , Britta Kluge , Sabine Heiland","doi":"10.1016/j.brainresprot.2003.11.004","DOIUrl":"10.1016/j.brainresprot.2003.11.004","url":null,"abstract":"<div><p>Magnetic resonance imaging (MRI) is applied in many studies on experimental cerebral ischemia in rodents to monitor the temporal evolution of ischemic damage. We report a protocol to evaluate the infarct size after middle cerebral artery occlusion with reperfusion (MCAO/R) in male Wistar rats. Imaging was performed with a 2.35 T scanner and we focused on diffusion-weighted imaging (DWI), T2-weighted imaging (T2WI) and postcontrast T1-weighted imaging (T1WI). We show the detailed procedure of volumetry, the contrast-to-noise ratio (CNR) and the intraindividual variability of infarct and hemispheric volumes at different reperfusion times. The presented method is of low variability if image contrast between ischemic and nonischemic tissue is very high, which is the case not only for all sequences at 8 and 12 h of reperfusion but also for DWI after 3 and 5 h of reperfusion. Furthermore, we describe the so-called mismatch region of lesion sizes depicted on DWI and postcontrast T1WI that suffers from cytotoxic edema but lacks contrast enhancement.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"12 3","pages":"Pages 172-179"},"PeriodicalIF":0.0,"publicationDate":"2004-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.11.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40835261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

Middle cerebral artery occlusion during MR-imaging: investigation of the hyperacute phase of stroke using a new in-bore occlusion model in rats 磁共振成像时大脑中动脉闭塞:用一种新的脑内闭塞模型研究大鼠脑卒中超急性期

Brain research. Brain research protocols Pub Date : 2004-02-01 DOI: 10.1016/j.brainresprot.2003.08.006

Tibo Gerriets , Erwin Stolz , Maureen Walberer , Clemens Müller , Alexander Kluge , Manfred Kaps , Marc Fisher , Georg Bachmann

{"title":"Middle cerebral artery occlusion during MR-imaging: investigation of the hyperacute phase of stroke using a new in-bore occlusion model in rats","authors":"Tibo Gerriets , Erwin Stolz , Maureen Walberer , Clemens Müller , Alexander Kluge , Manfred Kaps , Marc Fisher , Georg Bachmann","doi":"10.1016/j.brainresprot.2003.08.006","DOIUrl":"10.1016/j.brainresprot.2003.08.006","url":null,"abstract":"<div><p>Magnetic resonance imaging (MRI) provides insights into the dynamics of focal cerebral ischemia. Usually, experimental stroke is induced outside the magnet bore, preventing investigators from acquiring pre-ischemic images for later pixel-by-pixel comparisons and from studying the earliest changes in the hyperacute phase of ischemia. Herein, we introduce a new and easy to apply in-bore occlusion protocol based on the intraarterial embolization of ceramic macrospheres.</p><p><span>PE-50 tubing, filled with saline and six macrospheres (0.315–0.355 mm in diameter), was placed into the internal carotid artery (ICA) of anesthetized Sprague–Dawley rats. The animals were transferred into an MRI scanner (7.0 T) and baseline diffusion-weighted imaging (DWI) and T2-imaging was performed. Then the macrospheres were injected into the internal artery to occlude the </span>MCA. Post-ischemic DWI and T2-imaging was started immediately thereafter. The apparent diffusion coefficient (ADC) (a marker for cytotoxic brain edema) and T2-relaxation time (a marker for vasogenic brain edema) were determined in the ischemic lesions and compared to the unaffected hemisphere.</p><p>ADC significantly declined within the first 5–10 min after stroke onset. T2-relaxation time increased as early as at the first T2-imaging time-point (20–35 min after embolization). After 150 min of ischemia, the lesions covered 18.0±7.4% of the hemispheres. The model failed in one out of nine animals (11%).</p><p>This model allows MR-imaging from the initial minutes after permanent middle cerebral artery (MCA) occlusion. It does not permit reperfusion. This technique might provide information about the pathophysiological processes in the hyperacute phase of stroke.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"12 3","pages":"Pages 137-143"},"PeriodicalIF":0.0,"publicationDate":"2004-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.08.006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40835324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 54

Free colour illustrations in the online version of articles 免费彩色插图在文章的在线版本

Brain research. Brain research protocols Pub Date : 2004-02-01 DOI: 10.1016/S1385-299X(04)00008-X

引用次数: 0

EEG-dependent ERP recording: using TMS to increase the incidence of a selected pre-stimulus pattern 脑电依赖性ERP记录:使用经颅磁刺激增加选定的预刺激模式的发生率

Brain research. Brain research protocols Pub Date : 2004-02-01 DOI: 10.1016/j.brainresprot.2003.10.002

Gregory W Price

{"title":"EEG-dependent ERP recording: using TMS to increase the incidence of a selected pre-stimulus pattern","authors":"Gregory W Price","doi":"10.1016/j.brainresprot.2003.10.002","DOIUrl":"10.1016/j.brainresprot.2003.10.002","url":null,"abstract":"<div><p>EEG dependent event-related potential (ERP) recording (interactive ERP) is an extension to ERP paradigms whereby stimuli are initiated in response to a selected pattern of background EEG. This form of recording is critically dependent upon the incidence of the particular pattern of interest. We introduce here a process that modifies the EEG in a predictable manner so as to increase the incidence of a particular pattern. Transcranial magnetic stimulation (TMS) stimuli are applied in response to a selected pattern of pre-TMS activity, and the post-TMS response is characterized by the incidence of a defined pattern of EEG activity. Analysis of validation test results obtained with the TMS modification part of the process verifies an increased incidence of the response pattern after TMS stimuli, compared with placebo stimuli. The TMS modification procedure is then combined with interactive ERP recording in a two step process to affect the ERP response to sensory stimuli. The post-TMS pattern from the first step becomes the pre-stimulus pattern of the interactive ERP recording in the second step. The TMS modified interactive ERP (TMIERP) process is illustrated here using an auditory oddball paradigm. The amplitude of the P300 peak obtained using this process was significantly higher than that obtained using the standard auditory oddball paradigm.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"12 3","pages":"Pages 144-151"},"PeriodicalIF":0.0,"publicationDate":"2004-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.10.002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40835325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10