Name: measuring near plasma membrane and global intracellular calcium dynamics in astrocytes
Uploaded: 2009-04-26T23:27:00
Description: Watch this Scientific Journal Video about Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes at JoVE.com

Bu çalışma, Japonya Uehara Memorial Foundation (ES) yanı sıra Whitehall Vakfı, Ulusal Nörolojik Bozukluklar ve İnme ve Stein-Oppenheimer Endowment Ödülü (BSK) Enstitüsü tarafından desteklendi.

DENEYSEL PROSEDÜRLER Deneysel işlemin bir adım daha akıllıca bir şekilde aşağıda açıklanan iki ana bölümden oluşmaktadır. Bölüm 1: HAZIRLAMA hipokampal astrosit KÜLTÜRLER Kısaca, karışık, hipokampal astrosit nöron kültürleri iyi kurulmuş bir protokol 1,2,3 kullanılarak hazırlanmıştır. Prosedürü, sağlıklı kültürlü astrositler verim için optimize edilmiş. Aşağıda listelenen tüm işlem...

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Astrositler hücre içi kalsiyum yükselmeler görüntüler çok iyi kurulmuştur. Bu, kendiliğinden meydana gelen nöronal aktivite veya 11 astrosit yüzey reseptörleri aktive agonistleri uygulama tarafından tetiklenebilir. Bir önemli ve tartışmalı bir konu olmadığını astrosit hücre içi kalsiyum yükselmeleri, nöronlar 11, 12 reseptörleri aktive sinyal molekülleri serbest bırakılması tetikleyebilir. Yorum Haydon 7, 13 ve McCarthy 11 laboratuvarları tarafından vurgulanan bu g?...

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<td>VWR&#174; Micro Cover Slips, Round, No. 1</td>
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<td>VWR</td>
<td>48380-068</td>
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<td>Poly-D-lysine hydrobromide</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P0899</td>
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<td>Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane</td>
<td>Reagent</td>
<td>Sigma</td>
<td>L2020</td>
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<td>Earle&#x2019;s Balanced Salt Solution (EBSS) (1X), liquid</td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>14155-063</td>
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<td>Minimum Essential Medium (MEM) (1X), liquid Contains Earle's salts, but no L-glutamine or phenol red</td>
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<td>Invitrogen</td>
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<td>Penicillin-Streptomycin liquid</td>
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<td>Invitrogen</td>
<td>15140-122</td>
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<td>Sodium pyruvate solution</td>
<td>Reagent</td>
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<td>S8636</td>
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<td>HEPES solution 1 M</td>
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<td>N-2 Supplement (100X), liquid</td>
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<td>Invitrogen</td>
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<td>Horse Serum, Heat-Inactivated</td>
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<td>Invitrogen</td>
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<td>PAPAIN-022</td>
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<td>Worthington</td>
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<td>Neurobasal&#x2122; Medium (1X) Liquid without Phenol Red</td>
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<td>Invitrogen</td>
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<td>B-27 Serum-Free Supplement (50X), liquid</td>
<td>Reagent</td>
<td>Invitrogen</td>
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<td>Invitrogen</td>
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<td>Cell Strainers</td>
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<td>BD Biosciences</td>
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<td>P3911</td>
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<td>21108</td>
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<td>M2670</td>
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<td>H3375</td>
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<td>D-(+)-glucose</td>
<td>Reagent</td>
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<td>G7528</td>
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<td>Fluo-4, AM 1 mM solution in DMSO</td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>F-14217</td>
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<td>Pluronic&#174; F-127 20% solution in DMSO</td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>P-3000MP</td>
<td>&nbsp;</td>
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<td>Immersion Oil TYPE DF</td>
<td>Microscope</td>
<td>Cargille</td>
<td>16242</td>
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<td>Open chamber for 25 mm round coverslips, 100 &mu;l volume</td>
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<td>WARNER Instruments</td>
<td>64-0362 (RC-21BDW)</td>
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<td>P-2 platform for Series 20 chambers, non-heater</td>
<td>Tool</td>
<td>WARNER Instruments</td>
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measuring near plasma membrane and global intracellular calcium dynamics in astrocytes

Beyin glial hücreler içerir. Astrositler, glial hücre tipi, uzun nöronlar için pasif bir destekleyici bir rol sağlamak için bilinmektedir. Ancak, artan kanıtlar astrositler nöronlar ile fonksiyonel etkileşim yoluyla da aktif beyin fonksiyonu katılmak olabileceğini düşündürmektedir. Ancak, birçok temel yönleri astrosit biyoloji, tartışmalı, belirsiz ve / veya deneysel keşfedilmemiş kalır. Bir önemli konu astrositlerde hücre içi kalsiyum Transientlerin dinamikleri. Kalsiyum önemli bir ikinci haberci olarak iyi kurulmuş ve çünkü astrosit kalsiyum yükselmeler astrositler gelen vericileri serbest bırakılması tetikleyebilir ileri sürülmektedir çünkü bu geçerlidir. Ancak, yakın plazma membranı kalsiyum astrositler sinyal herhangi bir ayrıntılı ve tatmin edici bir açıklama olmamıştır. Toplam iç yansıma floresan (TIRF) mikroskopi canlı hücrelerinin plazma membranı yaklaşık 100 nm içinde önemli fizyolojik sinyalleme olaylarını analiz etmek için güçlü bir araçtır. Burada, TIRF mikroskobu kullanımı ve neredeyse aynı anda, plazma membranı ve küresel hücre içi kalsiyum dinamikleri yakın izlemek nasıl tarif. Bu yaklaşımın daha rafine ve sistematik bir uygulama astrosit kalsiyum sinyal kesin detaylar hakkında bilgi potansiyeline sahiptir. Astrosit kalsiyum dinamiklerinin ayrıntılı bir anlayış, nasıl, ne zaman ve neden astrositler ve nöron kalsiyum-bağımlı fonksiyonel etkileşimleri tabi anlamak için bir temel sağlayabilir.

Watch this Scientific Journal Video about Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes at JoVE.com

Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes

Biz toplam iç yansıma ve Epifloresans mikroskopi kullanılarak kültürlü astrositler membran ve küresel hücre içi kalsiyum dinamikleri yakın ölçmek için nasıl açıklar.

Astrositler Plazma Membran ve Küresel Hücre içi kalsiyum Dynamics civarında Ölçüm

The brain contains glial cells.  Astrocytes, a type of glial cell, have long been known to provide a passive supportive role to neurons. However, ...

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Biology

Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation

Tersinir biyotinilasyon Plazma Membran Protein endositik Oranları Ölçüm

Plasma membrane proteins are a large, diverse group of proteins comprised of receptors, ion channels, transporters and pumps.  Activity of these proteins ...

Measuring Fast Calcium Fluxes in Cardiomyocytes

Hızlı Kalsiyum Dekapanları kardiyomiyositlerde Ölçme

Cardiomyocytes have multiple Ca2+ fluxes of varying duration that work together to optimize function 1,2. Changes in Ca2+ activity in response to ...

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing (MTT)

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT

Çoklu Hedef İzleme (MTT) tarafından Plazma Membran Moleküler Difüzyon Haritalama

Our goal is to obtain a comprehensive description of molecular processes occurring at cellular membranes in different biological functions. We aim at ...

Intravital Microscopy for Imaging Subcellular Structures in Live Mice Expressing Fluorescent Proteins

Floresan Proteinler ifade Canlı Farelerde Görüntüleme hücrealtı Yapıların Intravital Mikroskopi

Here we describe a procedure to image subcellular structures in live rodents that is based on the use of confocal intravital microscopy. As a model organ, ...

Imaging Plasma Membrane Deformations With pTIRFM

PTIRFM ile görüntüleme Plazma Membran deformasyonlar

To gain novel insights into the dynamics of exocytosis, our group focuses on the changes in lipid bilayer shape that must be precisely regulated during ...

Functional Characterization of Na+/H+ Exchangers of Intracellular Compartments Using Proton-killing Selection to Express Them at the Plasma Membrane

Functional Characterization of Na+/H+ Exchangers of Intracellular Compartments Using Proton-killing Selection to Express Them at the Plasma Membrane

Na fonksiyonel karakterizasyonu<sup&gt; +</sup&gt; / H<sup&gt; +</sup&gt; Seçim Proton-öldürme Kullanma hücre içi Bölmeler ve Eşanjörleri Plazma Membran Them Express

Endosomal acidification is critical for a wide range of processes, such as protein recycling and degradation, receptor desensitization, and ...

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

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This method provides a fast approach for the determination of plasma membrane partitioning of any fluorescently-tagged peripherally-associated protein ...

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis

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Necroptosis is a programmed cell death pathway triggered by activation of receptor interacting protein kinase 3 (RIPK3), which phosphorylates and ...

Simultaneous Measurements of Intracellular Calcium and Membrane Potential in Freshly Isolated and Intact Mouse Cerebral Endothelium

Hücre içi kalsiyum ve taze izole ve olduğu gibi fare serebral endotel membran potansiyeli eşzamanlı ölçümler

Cerebral arteries and their respective microcirculation deliver oxygen and nutrients to the brain via blood flow regulation. Endothelial cells line the ...

Imaging Calcium Dynamics in Subpopulations of Mouse Pancreatic Islet Cells

Fare Pankreas Adacık Hücrelerinin Alt Popülasyonlarında Görüntüleme Kalsiyum Dinamiği

Pancreatic islet hormones regulate blood glucose homeostasis. Changes in blood glucose induce oscillations of cytosolic calcium in pancreatic islet cells ...