作者: 聲音刺耳 時(shí)間: 2025-3-21 23:49 作者: 同音 時(shí)間: 2025-3-22 01:59
Francis A. Gunther,Jane Davies Gunther pancreatic β-cells. The presynaptic capacitance measurements provide a unique alternative to measuring transmitter release and presynaptic endocytosis. Here, we describe this method at the calyx of Held in acute brain slices and provide a practical guide to obtaining high quality capacitance measur作者: fatty-acids 時(shí)間: 2025-3-22 08:07 作者: 浮夸 時(shí)間: 2025-3-22 11:44 作者: 使堅(jiān)硬 時(shí)間: 2025-3-22 14:49
Correction to: Clathrin-Mediated Endocytosis: Methods and Protocols,作者: 使堅(jiān)硬 時(shí)間: 2025-3-22 18:16 作者: archetype 時(shí)間: 2025-3-22 23:34
https://doi.org/10.1007/978-1-4615-8495-7ndamental to study the biochemistry and the physiology of the nervous system. This chapter describes the isolation and purification of intact synaptosomes from rodent brain tissue that can be used to further characterize synaptic structure and function and to examine the molecular mechanisms of neurotransmission.作者: 得意人 時(shí)間: 2025-3-23 02:29
Gerald A. Pollock,Wendell W. Kilgorethe use of FLP/FRT-mediated mitotic recombination to generate . having homozygous mutant eyes while the rest of their body is heterozygous. We then present a detailed protocol for assessing the consequences of these loss-of-function mutations on endocytosis in the photoreceptors of living fruit flies by recording electroretinograms.作者: 小歌劇 時(shí)間: 2025-3-23 08:53 作者: Amplify 時(shí)間: 2025-3-23 10:33 作者: Crayon 時(shí)間: 2025-3-23 14:48
Preparation of Synaptosomes from Mammalian Brain by Subcellular Fractionation and Gradient Centrifundamental to study the biochemistry and the physiology of the nervous system. This chapter describes the isolation and purification of intact synaptosomes from rodent brain tissue that can be used to further characterize synaptic structure and function and to examine the molecular mechanisms of neurotransmission.作者: trigger 時(shí)間: 2025-3-23 19:31 作者: BRAND 時(shí)間: 2025-3-23 23:20 作者: Between 時(shí)間: 2025-3-24 03:08
Cryo-Electron Tomography of the Mammalian Synapse, unstained, fully hydrated cellular structures at molecular resolution. The protocol focuses on purified synaptic terminals (“synaptosomes”), currently the most suitable preparation to analyze mammalian synaptic architecture by cryo-ET.作者: 裹住 時(shí)間: 2025-3-24 10:18 作者: 精密 時(shí)間: 2025-3-24 13:51
,Imaging “Hot-Wired” Clathrin-Mediated Endocytosis,d in our system. In this chapter, we describe in detail how to use the hot-wiring system to trigger endocytosis in human cell lines and how to image the vesicles that form using microscopy and finally, how to analyze those images.作者: 非秘密 時(shí)間: 2025-3-24 15:23 作者: 知道 時(shí)間: 2025-3-24 19:06 作者: 連詞 時(shí)間: 2025-3-25 01:10 作者: adhesive 時(shí)間: 2025-3-25 06:34
https://doi.org/10.1007/978-1-4613-9407-5cribe a methodology to follow adaptor-induced clathrin assembly in real-time using fluorescence microscopy on a facile model membrane assay system of .upported .emb.ane .ubes (SMrT). Results from such assays can be conveniently run through routine image analysis procedures to extract kinetic parameters of the clathrin assembly reaction.作者: Conspiracy 時(shí)間: 2025-3-25 11:18 作者: Generalize 時(shí)間: 2025-3-25 14:37 作者: 共同時(shí)代 時(shí)間: 2025-3-25 17:12
SMrT Assay for Real-Time Visualization and Analysis of Clathrin Assembly Reactions,cribe a methodology to follow adaptor-induced clathrin assembly in real-time using fluorescence microscopy on a facile model membrane assay system of .upported .emb.ane .ubes (SMrT). Results from such assays can be conveniently run through routine image analysis procedures to extract kinetic parameters of the clathrin assembly reaction.作者: mastoid-bone 時(shí)間: 2025-3-25 21:07
Francis A. Gunther,Jane Davies Gunthered by mRNA interference or dominant-negative mutant overexpression affect endocytosis long before cells are being assayed. Here, we describe simple experimental procedures to assay endocytosis along the cell cycle with minimal perturbations.作者: Hypopnea 時(shí)間: 2025-3-26 01:59
Glenn C. Miller,Richard G. Zepp,5)P. spatial distribution, and semi-quantification of PI(4,5)P. levels in the plasma membrane using fluorescence microscopy. Depending on the probe selected for lipid detection, this simple assay can be modified to study other plasmalemmal phospholipids and/or proteins.作者: 整潔 時(shí)間: 2025-3-26 04:46 作者: Libido 時(shí)間: 2025-3-26 09:57
Spatial and Temporal Aspects of Phosphoinositides in Endocytosis Studied in the Isolated Plasma Mem,5)P. spatial distribution, and semi-quantification of PI(4,5)P. levels in the plasma membrane using fluorescence microscopy. Depending on the probe selected for lipid detection, this simple assay can be modified to study other plasmalemmal phospholipids and/or proteins.作者: 飛鏢 時(shí)間: 2025-3-26 15:49
Annette E. McCann,D. Roy Cullimored in our system. In this chapter, we describe in detail how to use the hot-wiring system to trigger endocytosis in human cell lines and how to image the vesicles that form using microscopy and finally, how to analyze those images.作者: 我們的面粉 時(shí)間: 2025-3-26 19:43 作者: Lipohypertrophy 時(shí)間: 2025-3-26 22:10
Preparation of Synaptosomes from Mammalian Brain by Subcellular Fractionation and Gradient Centrifuommunication. Among other techniques, the isolation of nerve terminals [or synaptosomes (Whittaker et al. Biochem J, 90(2):293–303, 1964)] has been fundamental to study the biochemistry and the physiology of the nervous system. This chapter describes the isolation and purification of intact synaptos作者: BOON 時(shí)間: 2025-3-27 05:04
Probing Endocytosis During the Cell Cycle with Minimal Experimental Perturbation, during infection. Endocytosis activity is known to vary during the cell cycle, in particular during mitosis. Importantly, different experimental conditions can lead to opposite results and conclusions, thereby emphasizing the need for a careful design of protocols. For example, experiments using se作者: 可能性 時(shí)間: 2025-3-27 07:10
Assaying the Contribution of Membrane Tension to Clathrin-Mediated Endocytosis, Since the implementation of this methodology to the field of clathrin-mediated endocytosis (CME), this approach has revolutionized our molecular understanding of clathrin-driven cellular uptake. Conventional live cell microscopy approaches are used to determine the precise functions of specific pro作者: 為敵 時(shí)間: 2025-3-27 11:59
Identifying Small-Molecule Inhibitors of the Clathrin Terminal Domain,ncluding receptors for nutrients and signaling molecules, as well as synaptic vesicle reformation. Multiple genetic and chemical approaches have been developed to interfere with this process. However, many of these tools do not selectively block CME, for example by targeting components shared with c作者: 陳舊 時(shí)間: 2025-3-27 16:52 作者: FEMUR 時(shí)間: 2025-3-27 18:22 作者: giggle 時(shí)間: 2025-3-27 23:12 作者: Ptosis 時(shí)間: 2025-3-28 03:22 作者: Kernel 時(shí)間: 2025-3-28 07:35
Reconstitution of Clathrin Coat Disassembly for Fluorescence Microscopy and Single-Molecule Analysi recruitment of auxilin and Hsc70, an ATP-driven molecular clamp. Here, we describe the preparation of reagents and the single-particle fluorescence microscopy imaging assay in which we visualize directly the Hsc70-driven uncoating of synthetic clathrin coats or clathrin-coated vesicles.作者: HPA533 時(shí)間: 2025-3-28 10:54
Spatial and Temporal Aspects of Phosphoinositides in Endocytosis Studied in the Isolated Plasma Memembrane phospholipids have key regulatory functions in endocytosis and membrane traffic. I have previously described an in vitro assay based on the isolated, substrate-attached plasma membrane to study the spatial distribution and levels of phosphoinositides, in particular phosphatidylinositol-4,5-b作者: Ambiguous 時(shí)間: 2025-3-28 17:27 作者: BIPED 時(shí)間: 2025-3-28 21:55
Real-Time Monitoring of Clathrin Assembly Kinetics in a Reconstituted System,iously described an assay of reconstituting ccp assembly on sheets of basal plasma membranes. Here, we describe a workflow to adapt this system for monitoring the assembly of ccps over time using total internal reflection fluorescence (TIRF) microscopy.作者: 無(wú)節(jié)奏 時(shí)間: 2025-3-28 23:28
Stimulated Emission Depletion (STED) Imaging of Clathrin-Mediated Endocytosis in Living Cells,ular processes with unprecedented details. Here we describe how to image endocytic events at the plasma membrane of living cells using commercial (Leica, Abberior Instruments) or custom built STED microscopes.作者: NEX 時(shí)間: 2025-3-29 04:29
Measuring Clathrin-Coated Vesicle Formation with Single-Molecule Resolution,ty information, imaging experiments can be rendered quantitative, yielding insights into the stoichiometry and kinetics of the components of a molecular assembly. Here, we describe the experimental and analytical steps needed to study the assembly of clathrin-coated vesicles with single-molecule res作者: 可以任性 時(shí)間: 2025-3-29 07:53 作者: Euthyroid 時(shí)間: 2025-3-29 15:09
Using FM Dyes to Monitor Clathrin-Mediated Endocytosis in Primary Neuronal Culture,ne bound are intensely fluorescent, combined with the flexibility of different emission wavelengths make these dyes an excellent choice for investigating clathrin-mediated endocytosis, among other membrane trafficking and recycling pathways.作者: CANON 時(shí)間: 2025-3-29 16:18 作者: 大廳 時(shí)間: 2025-3-29 22:46
978-1-4939-9374-1Springer Science+Business Media, LLC, part of Springer Nature 2018作者: 十字架 時(shí)間: 2025-3-30 00:15 作者: 變形詞 時(shí)間: 2025-3-30 06:31 作者: ACRID 時(shí)間: 2025-3-30 09:49 作者: 金絲雀 時(shí)間: 2025-3-30 14:13 作者: 宴會(huì) 時(shí)間: 2025-3-30 17:18
Francis A. Gunther,Jane Davies Gunther during infection. Endocytosis activity is known to vary during the cell cycle, in particular during mitosis. Importantly, different experimental conditions can lead to opposite results and conclusions, thereby emphasizing the need for a careful design of protocols. For example, experiments using se作者: RALES 時(shí)間: 2025-3-30 23:20 作者: 巡回 時(shí)間: 2025-3-31 04:36
https://doi.org/10.1007/978-1-4613-9388-7ncluding receptors for nutrients and signaling molecules, as well as synaptic vesicle reformation. Multiple genetic and chemical approaches have been developed to interfere with this process. However, many of these tools do not selectively block CME, for example by targeting components shared with c作者: fetter 時(shí)間: 2025-3-31 07:59 作者: 協(xié)迫 時(shí)間: 2025-3-31 10:07 作者: uncertain 時(shí)間: 2025-3-31 16:02 作者: 創(chuàng)造性 時(shí)間: 2025-3-31 18:38
Gerald A. Pollock,Wendell W. Kilgorey not viable. Different approaches have been developed to circumvent this limitation, including resorting to mosaic model organisms. We here describe the use of FLP/FRT-mediated mitotic recombination to generate . having homozygous mutant eyes while the rest of their body is heterozygous. We then pr作者: Vertical 時(shí)間: 2025-4-1 01:32
Soil-parathion surface interactions, recruitment of auxilin and Hsc70, an ATP-driven molecular clamp. Here, we describe the preparation of reagents and the single-particle fluorescence microscopy imaging assay in which we visualize directly the Hsc70-driven uncoating of synthetic clathrin coats or clathrin-coated vesicles.作者: Defense 時(shí)間: 2025-4-1 02:08
Glenn C. Miller,Richard G. Zeppembrane phospholipids have key regulatory functions in endocytosis and membrane traffic. I have previously described an in vitro assay based on the isolated, substrate-attached plasma membrane to study the spatial distribution and levels of phosphoinositides, in particular phosphatidylinositol-4,5-b作者: slipped-disk 時(shí)間: 2025-4-1 06:56
https://doi.org/10.1007/978-1-4613-9407-5g this process, discrete sets of adaptor proteins recognize specific classes of membrane proteins, which recruit and assemble clathrin lattices on the membrane. An important determinant to the success of this vesicular transport reaction is the intrinsic ability of adaptors to polymerize clathrin on作者: Adulterate 時(shí)間: 2025-4-1 12:04
