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Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/S1385-299X(05)00055-3

引用次数: 0

In vivo preparation and identification of mitral cells in the main olfactory bulb of the mouse 小鼠主嗅球二尖瓣细胞的体内制备与鉴定

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.05.001

Thomas G. Mast, Edwin R. Griff

{"title":"In vivo preparation and identification of mitral cells in the main olfactory bulb of the mouse","authors":"Thomas G. Mast, Edwin R. Griff","doi":"10.1016/j.brainresprot.2005.05.001","DOIUrl":"10.1016/j.brainresprot.2005.05.001","url":null,"abstract":"<div><p>The mouse main olfactory bulb<span> (MOB) is commonly used as a mammalian model to study olfactory processing. The genetic techniques available with the mouse make its MOB a powerful model for analysis of neuronal circuitry. The mouse has been used as a mammalian model for all types of MOB neurons, but especially to study the activity of mitral cells. However, mouse mitral cell activity is most commonly studied in vitro. Therefore, we aimed to develop a protocol to record the activity of antidromically identified mitral cells in mouse in vivo. Currently, such a protocol does not exist. Using extracellular techniques, we report a protocol that is able to record neurons from all mouse MOB layers. Specifically, mitral cell single-units were identified by antidromic activation from the posterior piriform cortex<span>, and their spontaneous activity was recorded for more than 30 min. This protocol is stable enough to record from single-units while buprenorphine was applied both topically to the surface of the MOB and injected systemically.</span></span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 2","pages":"Pages 105-113"},"PeriodicalIF":0.0,"publicationDate":"2005-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.05.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25152087","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: 3

An examination of MS candidate genes identified as differentially regulated in multiple sclerosis plaque tissue, using absolute and comparative real-time Q-PCR analysis 使用绝对实时和比较实时Q-PCR分析，检测多发性硬化症斑块组织中被鉴定为差异调节的MS候选基因

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.04.003

L. Tajouri , A.S. Mellick , A. Tourtellotte , R.M. Nagra , L.R. Griffiths

{"title":"An examination of MS candidate genes identified as differentially regulated in multiple sclerosis plaque tissue, using absolute and comparative real-time Q-PCR analysis","authors":"L. Tajouri , A.S. Mellick , A. Tourtellotte , R.M. Nagra , L.R. Griffiths","doi":"10.1016/j.brainresprot.2005.04.003","DOIUrl":"10.1016/j.brainresprot.2005.04.003","url":null,"abstract":"<div><p><span>In our laboratory, we have developed methods in real-time detection and quantitative-polymerase chain reaction (Q-PCR) to analyse the relative levels of gene expression in post mortem brain tissues. We have then applied this method to examine differences in gene activity between normal white matter (NWM) and plaque tissue from multiple sclerosis (MS) patients. Genes were selected based on their association with pathology and through identification by previously conducted global gene expression analysis. Plaque tissue was obtained from secondary progressive (SP) patients displaying chronic active, as well as acute pathologies; while NWM from the same location was obtained from age- and sex-matched controls (normal patients). In this study, we used both SYBR Green I </span><em>supplementation</em> and commercially available mixes to assess both comparative and absolute levels of gene activity. The results of both methods compared favourably for four of the five genes examined (<em>P</em><span> < 0.05, Pearsons), while differences in gene expression between chronic active and acute pathologies were also identified. For example, a >50-fold increase in osteopontin (</span><em>Spp1</em><span>) and inositol 1-4-5 phosphate 3 kinase B (</span><em>Itpκb</em>) levels in acute plaques contrasted with the 5-fold or less increase in chronic active plaques (<em>P</em> < 0.05, unpaired <em>t</em> test). By contrast, there was no significant difference in the levels of the MS marker and calcium-dependent protease (Calpain, <em>Capns1</em>) in MS plaque tissue. In summary, Q-PCR analysis using SYBR Green I has allowed us to economically obtain what may be clinically significant information from small amounts of the CNS, providing an opportunity for further clinical investigations.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 2","pages":"Pages 79-91"},"PeriodicalIF":0.0,"publicationDate":"2005-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.04.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25120529","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: 30

Quantification of neurons expressing androgen receptor and volume estimation of the basolateral nuclear group of the canine amygdaloid body 雄激素受体表达神经元的定量和犬杏仁体基底外侧核群的体积估计

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.04.004

C. Jacobs, W. Van Den Broeck, P. Simoens

{"title":"Quantification of neurons expressing androgen receptor and volume estimation of the basolateral nuclear group of the canine amygdaloid body","authors":"C. Jacobs, W. Van Den Broeck, P. Simoens","doi":"10.1016/j.brainresprot.2005.04.004","DOIUrl":"10.1016/j.brainresprot.2005.04.004","url":null,"abstract":"<div><p><span>A protocol was developed for the stereological quantification of neurons expressing androgen receptor (AR) in the basolateral nuclear group (BNG) of the canine amygdaloid body. The Cavalieri method was used to estimate the BNG volume and the physical disector technique was applied for assessing the numerical densities and total numbers of both ordinary and AR-positive BNG neurons. The overall number of BNG neurons and the BNG volume were assessed on Nissl-stained sections, while AR was visualised using indirect </span>immunohistochemistry. The morphological differentiation between neurons, astrocytes, and oligodendrocytes in these immunohistochemical sections was hampered by the cytoplasmic localisation of AR in these cells. Therefore, an additional criterion was developed based on the nuclear diameters of these cells. With the cutoff value of 7.4 μm, a sensitivity of 97.7% and specificity of 97.6% were obtained. A negative correlation was found between the BNG volume and the numerical density of its neurons, implicating that a large BNG will not necessarily have a higher number of neurons. Therefore, the numerical density or BNG volume should always be assessed in addition to the total number of neurons, justifying the use of the physical disector instead of the fractionator technique in the present study. However, higher coefficients of error were obtained for the total number of neurons with the physical disector method because of the indirect measurement of cell numbers. Therefore, the precision of the estimates must be high enough when using the disector method to compensate the precision loss caused by this indirect calculation of the total cell number.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 2","pages":"Pages 92-104"},"PeriodicalIF":0.0,"publicationDate":"2005-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.04.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41014294","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: 9

A procedure for culturing astrocytes from white matter and the application of the siRNA technique for silencing the expression of their specific marker, S100A4 一种从白质中培养星形胶质细胞的方法，以及应用siRNA技术沉默其特异性标记物S100A4的表达

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.03.005

Elena N. Kozlova , Keizo Takenaga

{"title":"A procedure for culturing astrocytes from white matter and the application of the siRNA technique for silencing the expression of their specific marker, S100A4","authors":"Elena N. Kozlova , Keizo Takenaga","doi":"10.1016/j.brainresprot.2005.03.005","DOIUrl":"10.1016/j.brainresprot.2005.03.005","url":null,"abstract":"<div><p>White matter astrocytes have physiological functions which are distinct from those of astrocytes in gray matter. White matter becomes highly non-permissive to neurite growth<span><span><span> after injury, but the role of white matter astrocytes in this process is incompletely understood. Current protocols for making primary astroglial cultures are inadequate for exploring the specific properties of white matter astrocytes in vitro. We describe a procedure for obtaining cultures of white matter astrocytes from the rodent corpus callosum. In this procedure, we take advantage of our previous finding that white, but not gray matter astrocytes express the calcium-binding </span>protein S100A4. S100A4 expressing astrocytes are abundant in the corpus callosum, and we show that cultures, highly enriched in S100A4 expressing white matter astrocytes, can be reproducibly generated from this area. Key factors for successful cultures are (i) meticulous dissection of the corpus callosum from 4-day-old rats, and (ii) Percoll density gradient centrifugation to purify astrocytes. As a means of exploring the possible role of S100A4 in white matter astrocytes, we describe the use of the </span>siRNA technique to eliminate the expression of S100A4 in our in vitro system.</span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 2","pages":"Pages 59-65"},"PeriodicalIF":0.0,"publicationDate":"2005-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.03.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25152086","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: 13

Gene profiling of laser-microdissected brain regions and sub-regions 激光显微解剖脑区及亚区基因谱分析

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.04.002

Pietro Paolo Sanna, Alvin R. King, Lena D. van der Stap, Vez Repunte-Canonigo

引用次数: 16

Withdrawal syndrome from γ-hydroxybutyric acid (GHB) and 1,4-butanediol (1,4-BD) in Sardinian alcohol-preferring rats 撒丁岛嗜酒大鼠γ-羟基丁酸和1,4-丁二醇戒断综合征

Brain research. Brain research protocols Pub Date : 2005-07-01 DOI: 10.1016/j.brainresprot.2005.04.001

Mauro A.M. Carai , Lawrence S. Quang , Sergio Atzeri , Carla Lobina , Paola Maccioni , Alessandro Orrù , Gian Luigi Gessa , Timothy J. Maher , Giancarlo Colombo

{"title":"Withdrawal syndrome from γ-hydroxybutyric acid (GHB) and 1,4-butanediol (1,4-BD) in Sardinian alcohol-preferring rats","authors":"Mauro A.M. Carai , Lawrence S. Quang , Sergio Atzeri , Carla Lobina , Paola Maccioni , Alessandro Orrù , Gian Luigi Gessa , Timothy J. Maher , Giancarlo Colombo","doi":"10.1016/j.brainresprot.2005.04.001","DOIUrl":"https://doi.org/10.1016/j.brainresprot.2005.04.001","url":null,"abstract":"<div><p><span>γ-hydroxybutyric acid (GHB) and its precursors, 1,4-butanediol (1,4-BD) and γ-butyrolactone (GBL), are recreational drugs widely abused in the US, Europe and Australasia. A severe withdrawal syndrome from GHB, 1,4-BD and GBL has been increasingly documented over the last years, necessitating the development of a reliable animal model for investigations of potential therapeutic approaches. The present study describes the induction and occurrence of audiogenic seizures as a sign of withdrawal from GHB and 1,4-BD in selectively bred Sardinian alcohol-preferring (sP) rats, treated with escalating doses of GHB (1.5–3.5 g/kg, twice daily; i.g.) or 1,4-BD (500–1000 mg/kg, twice daily; i.g.) for 9 consecutive days. Acute administration of the selective GABA</span><sub>B</sub><span> receptor antagonist, SCH 50911, dramatically increased seizure occurrence. We propose that the inherent sensitivity of sP rats to different GHB-associated responses may have contributed to the unraveling of a phenomenon which was otherwise not recognizable in other rat strains.</span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 2","pages":"Pages 75-78"},"PeriodicalIF":0.0,"publicationDate":"2005-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.04.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"92013382","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: 7

Differences in the magnitude of long-term potentiation produced by theta burst and high frequency stimulation protocols matched in stimulus number 波爆发和高频刺激方案产生的长期增强幅度的差异在刺激数量上是一致的

Brain research. Brain research protocols Pub Date : 2005-05-01 DOI: 10.1016/j.brainresprot.2005.02.003

Ruben V. Hernandez , Mary M. Navarro , Ward A. Rodriguez , Joe L. Martinez Jr. , Richard G. LeBaron

{"title":"Differences in the magnitude of long-term potentiation produced by theta burst and high frequency stimulation protocols matched in stimulus number","authors":"Ruben V. Hernandez , Mary M. Navarro , Ward A. Rodriguez , Joe L. Martinez Jr. , Richard G. LeBaron","doi":"10.1016/j.brainresprot.2005.02.003","DOIUrl":"10.1016/j.brainresprot.2005.02.003","url":null,"abstract":"<div><p>Theta-burst stimulation (TBS: four pulses at 100 Hz repeated with 200 ms inter-burst-intervals) and another commonly used high-frequency stimulation protocol (HFS: 1 s burst of equally spaced pulses at 100 Hz) were compared for the magnitude of LTP produced in rat hippocampal slices. The total number of pulses applied during tetanus (TET) was either 40, 100, 200, or 300. In a conventional analysis of the last 10 min of the post-TET period, a two-way ANOVA revealed no difference either in LTP of the field excitatory post-synaptic potential (fEPSP) between TBS and HFS or differences across pulse number at 40, 100, or 200 pulses. At 300 pulses, there was a significant main effect by pulse number but not by protocol. A linear regression analysis showed that stimulation protocol accounted for only about 10% of the change in magnitude while pulse number contributed to 30% of the change. However, when an extended analysis of the same data was performed across the entire post-TET period with a repeated-measure ANOVA, a small but persistent increase in TBS over HFS at 200 pulses was significant. A difference between TBS and HFS at 300 pulses that occurred only during the early phase of LTP was also significant. These results suggest that, over a range of stimuli, the number of pulses in an induction protocol, rather than the pattern of stimulation, determines the magnitude of late phase LTP, while TBS produces greater potentiation than HFS in the early phase of LTP with higher TET number.</p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 1","pages":"Pages 6-13"},"PeriodicalIF":0.0,"publicationDate":"2005-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.02.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25096126","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: 55

A sequential fluorescence method for neurotransmitter-specific retrograde tracing in the central nervous system of the rat; utilizing True Blue and immunohistochemistry in combination with computer-assisted photography 序列荧光法用于大鼠中枢神经递质特异性逆行示踪利用True Blue和免疫组织化学结合计算机辅助摄影

Brain research. Brain research protocols Pub Date : 2005-05-01 DOI: 10.1016/j.brainresprot.2005.03.001

Anders Nylén , Bengt Larsson , Gunnar Skagerberg

{"title":"A sequential fluorescence method for neurotransmitter-specific retrograde tracing in the central nervous system of the rat; utilizing True Blue and immunohistochemistry in combination with computer-assisted photography","authors":"Anders Nylén , Bengt Larsson , Gunnar Skagerberg","doi":"10.1016/j.brainresprot.2005.03.001","DOIUrl":"10.1016/j.brainresprot.2005.03.001","url":null,"abstract":"<div><p><span>Aiming to map the distribution of spinally projecting, hypothalamic neurons containing neuronal nitric oxide synthase (nNOS), True Blue (TB) is injected into the rat spinal cord. After survival times of 7–14 days the animals are anaesthetized and perfused transcardially with a solution containing paraformaldehyde and sucrose. After dissection, the injection site is further fixed for 4–8 h, cut in a </span>cryostat<span>, and documented by computer-assisted digital photography. The brain region of interest is fixed for 4 h, rinsed in phosphate buffer for 48 h, sectioned, and photographically documented utilizing filter settings for visualization of TB. The brain sections are then immunohistochemically processed using a primary antibody against nNOS and a Texas Red (TR)-labelled secondary antibody and once again photographically documented, now using filter settings for visualization of TB and TR, respectively. Utilizing the Photoshop program, the TB containing cells can then be exactly aligned and the presence of TB and/or TR fluorescence in the same cell bodies are evaluated. This method for neurotransmitter-specific retrograde tracing derives its high sensitivity from the optimization of fixation/rinsing parameters, the use of appropriate fluorophores, and sequential digital microphotography.</span></p></div>","PeriodicalId":79477,"journal":{"name":"Brain research. Brain research protocols","volume":"15 1","pages":"Pages 30-37"},"PeriodicalIF":0.0,"publicationDate":"2005-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.brainresprot.2005.03.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25096129","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: 1

Themes and Topics 主题及专题

Brain research. Brain research protocols Pub Date : 2005-05-01 DOI: 10.1016/S1385-299X(05)00041-3

引用次数: 0