Brain research. Brain research protocols最新文献_第9页

Measuring membrane potential and electric field of brainstem neurons in vitro by confocal microscopy 用共聚焦显微镜测定体外脑干神经元膜电位和电场

Brain research. Brain research protocols Pub Date : 2004-06-01 DOI: 10.1016/j.brainresprot.2004.02.002

J Ma, F.Z Cui

引用次数: 2

A combined in situ hybridization and RT-PCR method to detect spatial and temporal patterns of Noggin gene expression in embryonic and postnatal rat hippocampus 采用原位杂交和RT-PCR相结合的方法检测胚胎和出生后大鼠海马中Noggin基因表达的时空格局

Brain research. Brain research protocols Pub Date : 2004-06-01 DOI: 10.1016/j.brainresprot.2004.01.005

Xiaotang Fan , Haiwei Xu , Yunjian Huang , Wenqin Cai

{"title":"A combined in situ hybridization and RT-PCR method to detect spatial and temporal patterns of Noggin gene expression in embryonic and postnatal rat hippocampus","authors":"Xiaotang Fan , Haiwei Xu , Yunjian Huang , Wenqin Cai","doi":"10.1016/j.brainresprot.2004.01.005","DOIUrl":"10.1016/j.brainresprot.2004.01.005","url":null,"abstract":"<div><p>Recent studies indicate that Noggin<span><span> not only plays an important role in the early development of the nervous system, but may also plays a role in postnatal central nervous system (CNS) development. In this study, we examined the relative levels and localization of Noggin mRNA in the hippocampus of rats of different developmental stages with reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization<span>. RT-PCR showed that the peak level of expression of Noggin mRNA was observed at embryonic day 13 (E13), subsequently gradually declined at 1–3 months (P1-3M) postnatal, and was detected only at a low level at P18M. In situ hybridization revealed that at embryonic stages, Noggin mRNA was localized throughout all hippocampal regions, whereas at early postnatal ages, Noggin mRNA was primarily localized in the anterior subiculum. At later postnatal ages, Noggin mRNA expression was obviously observed in the </span></span>dentate gyrus and in the CA1-CA3 pyramidal cell layers. Taken together, our results demonstrate that Noggin is expressed in embryonic and postnatal hippocampus, and that the temporal and spatial patterns of its expression is developmentally regulated.</span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 2","pages":"Pages 99-105"},"PeriodicalIF":0.0,"publicationDate":"2004-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2004.01.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24544155","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

Long double-stranded RNA-mediated RNA interference as a tool to achieve site-specific silencing of hypothalamic neuropeptides 长双链RNA介导的RNA干扰作为实现下丘脑神经肽位点特异性沉默的工具

Brain research. Brain research protocols Pub Date : 2004-06-01 DOI: 10.1016/j.brainresprot.2004.03.003

Aditi Bhargava , Mary F Dallman , David Pearce , SuJean Choi

{"title":"Long double-stranded RNA-mediated RNA interference as a tool to achieve site-specific silencing of hypothalamic neuropeptides","authors":"Aditi Bhargava , Mary F Dallman , David Pearce , SuJean Choi","doi":"10.1016/j.brainresprot.2004.03.003","DOIUrl":"10.1016/j.brainresprot.2004.03.003","url":null,"abstract":"<div><p><span><span>RNA interference (RNAi) has become a popular tool to silence gene expression in a variety of in vitro and in vivo systems. However, it has met with limited success in inhibiting gene expression in adult mammals. Here we demonstrate that long double-stranded RNA (dsRNA) can be used to create a “site-specific”, transient knockdown of genes in a fashion that is phenotypically akin to genetically manipulated organisms. Corticotropin-releasing factor (CRF) and arginine vasopressin<span> (AVP) that regulate a variety of physiological processes including the hypothalamic–pituitary–adrenal axis (HPA axis), energy and water homeostasis were used as model systems. Stereotaxic injections of dsRNA against CRF and AVP in the PVN specifically abolished the expression of these genes in the PVN leaving expression in other loci intact. Control dsRNA did not affect CRF or AVP expression in any brain region, suggesting that dsRNA did not shut down global protein synthesis. ANOVA showed significant main effects of silencing of CRF on dampening of the stress-activated release of </span></span>adrenocorticotrophin hormone (ACTH) (</span><em>F</em><sub>(2,7)</sub>=4.87; <em>p</em><0.047). Silencing of AVP resulted in increased water consumption, increased urine output and decreased urine osmolality as compared to control dsRNA-treated rats. Furthermore, dsRNA had no obvious deleterious effects on body weight or food consumption, variables considered essential in ruling out adverse physiologic effects in animal models. Thus, using long dsRNA, we were able to ascertain site-specific roles of CRF and AVP in adult rats without any developmental compensation and in a wild-type background.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 2","pages":"Pages 115-125"},"PeriodicalIF":0.0,"publicationDate":"2004-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2004.03.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24544157","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: 42

An in vivo method for recording single unit activity in lumbar spinal cord in mice anesthetized with a volatile anesthetic 用挥发性麻醉剂麻醉小鼠腰椎脊髓单单位活动的体内记录方法

Brain research. Brain research protocols Pub Date : 2004-06-01 DOI: 10.1016/j.brainresprot.2004.03.002

Jason M Cuellar , Joseph F Antognini , Earl Carstens

引用次数: 12

Completion by the 10th generation of the bidirectional selective breeding of GHB-sensitive and GHB-resistant rats 完成第10代ghb敏感大鼠和ghb抗性大鼠的双向选择性育种

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2004.02.001

Carla Lobina , Giancarlo Colombo , Gian Luigi Gessa , Mauro A.M. Carai

引用次数: 7

Self-injury behaviour induced by intraplantar carrageenan infiltration: a model of tonic nociception 足底卡拉胶浸润诱导的自伤行为:强直性伤害感觉模型

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2004.01.001

Alberto López-Avila , Gabriela Rodrı́guez-Manzo , Ulises Coffeen , Rosendo del Ángel , Francisco Pellicer

引用次数: 8

Themes and Topics 主题及专题

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/S1385-299X(04)00032-7

引用次数: 0

An intravital microscopy study of radiation-induced changes in permeability and leukocyte–endothelial cell interactions in the microvessels of the rat pia mater and cremaster muscle 活体显微镜研究辐射诱导大鼠脑膜和肌微血管通透性变化和白细胞-内皮细胞相互作用

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2003.11.005

M.Waleed Gaber , Hong Yuan , John T Killmar , Michael D Naimark , Mohammad F Kiani , Thomas E Merchant

{"title":"An intravital microscopy study of radiation-induced changes in permeability and leukocyte–endothelial cell interactions in the microvessels of the rat pia mater and cremaster muscle","authors":"M.Waleed Gaber , Hong Yuan , John T Killmar , Michael D Naimark , Mohammad F Kiani , Thomas E Merchant","doi":"10.1016/j.brainresprot.2003.11.005","DOIUrl":"10.1016/j.brainresprot.2003.11.005","url":null,"abstract":"<div><p><span>Using intravital microscopy and a closed window method, we measured irradiation-induced changes in the vascular permeability and cell interactions in microcirculation networks of the rat pia mater; the same effects were monitored in the </span>cremaster muscle as a control. The closed cranial window has many advantages, including long-term direct visualization of microcirculation. The method allows for repeated testing of the same vessel or network, thereby reducing variability. The method also allows for measurement of permeability changes and the accompanying leukocyte–endothelial cell interactions in the same network or vessel, which permits correlative studies of these phenomena. However, this method is not without challenges. The optical conditions are difficult, because the brain is three-dimensional and its parenchyma is more complex than the thinner, flatter peripheral tissues. To overcome this limitation, we performed a dynamic background subtraction. The background is dynamically related to vessel intensity, and changes in intensity were determined by eliminating the effects of neighboring and underlying vessels. We applied this method to studying the effects of ionizing radiation on the blood–brain barrier (BBB) permeability and cell interactions and the modulation of these effects by anti-ICAM-1 antibodies. Our results demonstrate that this method is sensitive to changes in these properties of the BBB.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 1-10"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.11.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451991","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: 68

Gene expression profiles derived from single cells in human postmortem brain 人类死后大脑单细胞的基因表达谱

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2003.12.003

Lixia Lu , Frauke Neff , Zhou Dun , Bernhard Hemmer , Wolfgang H Oertel , Jürgen Schlegel , Andreas Hartmann

{"title":"Gene expression profiles derived from single cells in human postmortem brain","authors":"Lixia Lu , Frauke Neff , Zhou Dun , Bernhard Hemmer , Wolfgang H Oertel , Jürgen Schlegel , Andreas Hartmann","doi":"10.1016/j.brainresprot.2003.12.003","DOIUrl":"10.1016/j.brainresprot.2003.12.003","url":null,"abstract":"<div><p>The study of postmortem human brain tissue remains the basis for the understanding of many CNS disorders and to verify data obtained in experimental studies. So far, however, gene expression profiling in cellular sub-populations derived from human postmortem brain was hampered by several technical drawbacks. Here, we describe a method that allows the generation of mRNA expression profiles from single neurons. Dopaminergic neurons from different midbrain areas including substantia nigra, central gray substance and ventral tegmental area were identified and isolated by immuno-laser capture microscopy (LCM). Expression profiles were generated from microdissected cells using a modified RNA fingerprinting protocol. Using this approach, we were able to generate specific RNA fingerprints at a high resolution from phenotype-specific single neurons. Polymorphic fragments were isolated from gels and differential gene expression was confirmed by real-time PCR using gene-specific primer pairs and hybridization probes. The method described here is easy to use and reliable for profiling gene expression at the single cell level in human postmortem brain. It could therefore be valuable to open new insights into the molecular pathogenesis of CNS disorders.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 18-25"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.12.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451993","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: 23

An optimized triphenyltetrazolium chloride method for identification of cerebral infarcts 一种优化的三苯四氮氯化铵鉴别脑梗死的方法

Brain research. Brain research protocols Pub Date : 2004-04-01 DOI: 10.1016/j.brainresprot.2003.12.001

Chintamani Narasinh Joshi , Swatantra Kumar Jain , Puvvada Sri Ramachandra Murthy

{"title":"An optimized triphenyltetrazolium chloride method for identification of cerebral infarcts","authors":"Chintamani Narasinh Joshi , Swatantra Kumar Jain , Puvvada Sri Ramachandra Murthy","doi":"10.1016/j.brainresprot.2003.12.001","DOIUrl":"10.1016/j.brainresprot.2003.12.001","url":null,"abstract":"<div><p><span><span>2,3,5-Triphenyltetrazolium chloride (TTC) staining is a convenient procedure for detection of brain infarcts but no standardized procedure is available. We report here an optimized and economic procedure of staining with TTC. Rats were subjected to reversible </span>middle cerebral artery (MCA) occlusion (2-h ischemia and 24-h reperfusion). At the end of reperfusion, brain was isolated and sliced rostro-caudally into serial 2-mm-thick slices. Sets of three serial slices from each brain were incubated for 30 min at 37 °C in three different concentrations of TTC—the first slice of the set in 1%, the second in 0.05% and the third in 0.1% TTC—in phosphate-buffered saline. Staining characteristics, optical density (OD) and infarct size were compared between juxtaposing cut surfaces of the slices stained with the three concentrations of TTC. After the first use, 0.05% TTC solution was stored at 4–8 °C and reused on the same day or on subsequent days. TTC at 0.05% concentration provided high contrast staining with clear demarcation between normal and infarct tissue. The infarct size in 0.05% TTC-stained slices correlated well with that in 0.1% TTC (</span><em>r</em>=0.92)- and 1% TTC (<em>r</em><span><span>=0.93)-stained preparations. ‘Nonspecific’ staining of corpus callosum and the </span>anterior commissures was minimal with the method. Once-used 0.05% TTC solution could be stored at 4–8 °C and reused. In conclusion, staining with 0.05% TTC provided improved delineation of brain infarcts, reduced ‘nonspecific’ staining of white matter and the infarct size correlated well with that measured after 1% TTC staining; the method also reduces the costs to 1/20.</span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"13 1","pages":"Pages 11-17"},"PeriodicalIF":0.0,"publicationDate":"2004-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2003.12.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24451992","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: 165