Brain cell biology最新文献_第2页

eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications. eNpHR:一种用于光遗传学应用的碱单胞菌紫红质。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-08-02 DOI: 10.1007/s11068-008-9027-6

Viviana Gradinaru, Kimberly R Thompson, Karl Deisseroth

{"title":"eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications.","authors":"Viviana Gradinaru, Kimberly R Thompson, Karl Deisseroth","doi":"10.1007/s11068-008-9027-6","DOIUrl":"https://doi.org/10.1007/s11068-008-9027-6","url":null,"abstract":"Temporally precise inhibition of distinct cell types in the intact nervous system has been enabled by the microbial halorhodopsin NpHR, a fast light-activated electrogenic Cl(-) pump. While neurons can be optically hyperpolarized and inhibited from firing action potentials at moderate NpHR expression levels, we have encountered challenges with pushing expression to extremely high levels, including apparent intracellular accumulations. We therefore sought to molecularly engineer NpHR to achieve strong expression without these cellular side effects. We found that high expression correlated with endoplasmic reticulum (ER) accumulation, and that under these conditions NpHR colocalized with ER proteins containing the KDEL ER retention sequence. We screened a number of different putative modulators of membrane trafficking and identified a combination of two motifs, an N-terminal signal peptide and a C-terminal ER export sequence, that markedly promoted membrane localization and ER export defined by confocal microscopy and whole-cell patch clamp. The modified NpHR displayed increased peak photocurrent in the absence of aggregations or toxicity, and potent optical inhibition was observed not only in vitro but also in vivo with thalamic single-unit recording. The new enhanced NpHR (eNpHR) allows safe, high-level expression in mammalian neurons, without toxicity and with augmented inhibitory function, in vitro and in vivo.","PeriodicalId":72445,"journal":{"name":"Brain cell biology","volume":" ","pages":"129-39"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11068-008-9027-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27576218","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 476

Real-time monitoring of cyclic nucleotide signaling in neurons using genetically encoded FRET probes. 利用基因编码FRET探针实时监测神经元中的环核苷酸信号。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-10-22 DOI: 10.1007/s11068-008-9035-6

Pierre Vincent, Nicolas Gervasi, Jin Zhang

引用次数: 20

Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP. 使用改进的非辐射YFP对单个树突棘进行高灵敏度和定量的FRET-FLIM成像。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-05-30 DOI: 10.1007/s11068-008-9024-9

Hideji Murakoshi, Seok-Jin Lee, Ryohei Yasuda

{"title":"Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP.","authors":"Hideji Murakoshi, Seok-Jin Lee, Ryohei Yasuda","doi":"10.1007/s11068-008-9024-9","DOIUrl":"https://doi.org/10.1007/s11068-008-9024-9","url":null,"abstract":"Two-photon fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue. We evaluated and optimized the FRET pair of mEGFP (monomeric EGFP with the A206K mutation) and REACh (non-radiative YFP variants) for TPFLIM. We characterized several mutants of REACh in terms of their \"darkness,\" and their ability to act as a FRET acceptor for mEGFP in HeLa cells and hippocampal neurons. Since the commonly used monomeric mutation A206K increases the brightness of REACh, we introduced a different monomeric mutation (F223R) which does not affect the brightness. Also, we found that the folding efficiency of original REACh, as measured by the fluorescence lifetime of a mEGFP-REACh tandem dimer, was low and variable from cell to cell. Introducing two folding mutations (F46L, Q69M) into REACh increased the folding efficiency by approximately 50%, and reduced the variability of FRET signal. Pairing mEGFP with the new REACh (super-REACh, or sREACh) improved the signal-to-noise ratio compared to the mEGFP-mRFP or mEGFP-original REACh pair by approximately 50%. Using this new pair, we demonstrated that the fraction of actin monomers in filamentous and globular forms in single dendritic spines can be quantitatively measured with high sensitivity. Thus, the mEGFP-sREACh pair is suited for quantitative FRET measurement by TPFLIM, and enables us to measure protein-protein interactions in individual dendritic spines in brain slices with high sensitivity.","PeriodicalId":72445,"journal":{"name":"Brain cell biology","volume":" ","pages":"31-42"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11068-008-9024-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27468305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 140

A heterogeneous "resting" pool of synaptic vesicles that is dynamically interchanged across boutons in mammalian CNS synapses. 在哺乳动物中枢神经系统突触中，一种异质的突触囊泡“静息”池在钮扣间动态交换。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-10-22 DOI: 10.1007/s11068-008-9030-y

Tomas Fernandez-Alfonso, Timothy A Ryan

引用次数: 91

Reporting neural activity with genetically encoded calcium indicators. 用基因编码的钙指标报告神经活动。

Brain cell biology Pub Date : 2008-08-01 DOI: 10.1007/s11068-008-9029-4

S Andrew Hires, Lin Tian, Loren L Looger

引用次数: 138

Optimizing the spatial resolution of Channelrhodopsin-2 activation. 通道视紫红质-2活化的空间分辨率优化。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-07-25 DOI: 10.1007/s11068-008-9025-8

Philipp Schoenenberger, Asa Grunditz, Tobias Rose, Thomas G Oertner

{"title":"Optimizing the spatial resolution of Channelrhodopsin-2 activation.","authors":"Philipp Schoenenberger, Asa Grunditz, Tobias Rose, Thomas G Oertner","doi":"10.1007/s11068-008-9025-8","DOIUrl":"https://doi.org/10.1007/s11068-008-9025-8","url":null,"abstract":"Over the past few years, the light-gated cation channel Channelrhodopsin-2 (ChR2) has seen a remarkable diversity of applications in neuroscience. However, commonly used wide-field illumination provides poor spatial selectivity for cell stimulation. We explored the potential of focal laser illumination to map photocurrents of individual neurons in sparsely transfected hippocampal slice cultures. Interestingly, the best spatial resolution of photocurrent induction was obtained at the lowest laser power. By adjusting the light intensity to a neuron's spike threshold, we were able to trigger action potentials with a spatial selectivity of less than 30 microm. Experiments with dissociated hippocampal cells suggested that the main factor limiting the spatial resolution was ChR2 current density rather than scattering of the excitation light. We conclude that subcellular resolution can be achieved only in cells with a high ChR2 expression level and that future improved variants of ChR2 are likely to extend the spatial resolution of photocurrent induction to the level of single dendrites.","PeriodicalId":72445,"journal":{"name":"Brain cell biology","volume":" ","pages":"119-27"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11068-008-9025-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27561485","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: 50

Optogenetic probes. 光遗传学探针。

Brain cell biology Pub Date : 2008-08-01 DOI: 10.1007/s11068-008-9037-4

Ryohei Yasuda, George J Augustine

引用次数: 3

Imaging synaptic inhibition throughout the brain via genetically targeted Clomeleon. 成像突触抑制整个大脑通过基因靶向Clomeleon。

Brain cell biology Pub Date : 2008-08-01 Epub Date: 2008-10-11 DOI: 10.1007/s11068-008-9031-x

Ken Berglund, Wolfram Schleich, Hong Wang, Guoping Feng, William C Hall, Thomas Kuner, George J Augustine

{"title":"Imaging synaptic inhibition throughout the brain via genetically targeted Clomeleon.","authors":"Ken Berglund, Wolfram Schleich, Hong Wang, Guoping Feng, William C Hall, Thomas Kuner, George J Augustine","doi":"10.1007/s11068-008-9031-x","DOIUrl":"https://doi.org/10.1007/s11068-008-9031-x","url":null,"abstract":"Here we survey a molecular genetic approach for imaging synaptic inhibition. This approach is based on measuring intracellular chloride concentration ([Cl(-)](i)) with the fluorescent chloride indicator protein, Clomeleon. We first describe several different ways to express Clomeleon in selected populations of neurons in the mouse brain. These methods include targeted viral gene transfer, conditional expression controlled by Cre recombination, and transgenesis based on the neuron-specific promoter, thy1. Next, we evaluate the feasibility of using different lines of thy1::Clomeleon transgenic mice to image synaptic inhibition in several different brain regions: the hippocampus, the deep cerebellar nuclei (DCN), the basolateral nucleus of the amygdala, and the superior colliculus (SC). Activation of hippocampal interneurons caused [Cl(-)](i) to rise transiently in individual postsynaptic CA1 pyramidal neurons. [Cl(-)](i) increased linearly with the number of electrical stimuli in a train, with peak changes as large as 4 mM. These responses were largely mediated by GABA receptors because they were blocked by antagonists of GABA receptors, such as GABAzine and bicuculline. Similar responses to synaptic activity were observed in DCN neurons, amygdalar principal cells, and collicular premotor neurons. However, in contrast to the hippocampus, the responses in these three regions were largely insensitive to antagonists of inhibitory neurotransmitter receptors. This indicates that synaptic activity can also cause Cl(-) influx through alternate pathways that remain to be identified. We conclude that Clomeleon imaging permits non-invasive, spatiotemporally precise recordings of [Cl(-)](i) in a large variety of neurons, and provides new opportunities for imaging synaptic inhibition and other forms of neuronal chloride signaling.","PeriodicalId":72445,"journal":{"name":"Brain cell biology","volume":" ","pages":"101-18"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11068-008-9031-x","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27724322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 59

Brain Cell Meetings 脑细胞会议

Brain cell biology Pub Date : 2008-01-01 DOI: 10.1007/s11068-009-9041-3

引用次数: 0

Welcome to Brain Cell Biology! 欢迎来到脑细胞生物学!

Brain cell biology Pub Date : 2007-03-10 DOI: 10.1007/S11068-006-9006-8

G. Augustine

引用次数: 0