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Related Concept Videos

Colloidal precipitates01:09

Colloidal precipitates

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The high insolubility of some precipitates can result in an unfavorable relative supersaturation. This can lead to colloidal particles with a large surface-to-mass ratio, where adsorption is promoted. For instance, in the precipitation of silver chloride, silver ions are adsorbed on the surface of the colloidal particles, forming a primary layer. This layer attracts ions of opposite charge (such as nitrate ions), forming a diffuse secondary layer of adsorbed ions. This electric double layer...
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The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
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Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
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Related Experiment Video

Updated: May 21, 2025

In Vitro Aggregation Assays Using Hyperphosphorylated Tau Protein
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Intracellular anionic substances cause tau liquid-liquid phase separation.

Kazuki Muguruma1, Tetsuya Takahashi2, Yuichiro Tagane1

  • 1Department of Clinical Neuroscience and Therapeutics, Hiroshima University Graduate School of Biomedical and Health Sciences, 1-2-3, Kasumi, Minami-ku, Hiroshima-shi, Hiroshima, 734-8551, Japan.

Biochemical and Biophysical Research Communications
|March 19, 2025
PubMed
Summary

Negatively charged substances and vesicles in cells can trigger tau protein liquid-liquid phase separation, a key step in neurodegenerative disease aggregation. This finding offers new therapeutic targets for tauopathies like Alzheimer's disease.

Keywords:
Alzheimer's diseaseAnionic substancesLiquid-liquid phase separationTau protein

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Cell Biology

Background:

  • Tau protein aggregation is central to neurodegenerative diseases such as Alzheimer's disease.
  • Liquid-liquid phase separation (LLPS) is an early mechanism in tau aggregation.
  • In vitro studies show tau forms liquid droplets with heparin, but in vivo evidence is lacking.

Purpose of the Study:

  • To investigate cellular conditions that induce tau liquid droplet formation in vivo.
  • To explore the role of anionic substances and intracellular vesicles in tau phase separation.
  • To understand the transition of tau droplets from liquid to solid states.

Main Methods:

  • Tau-expressing human embryonic kidney 293T cells were treated with anionic substances or agents inducing vesicle accumulation.
  • Niemann-Pick disease type C1 protein suppression and negatively charged dextran introduction were employed.
  • Tau-positive puncta formation and their dynamic state (liquid vs. solid) were observed over time.

Main Results:

  • Suppression of Niemann-Pick C1 or introduction of anionic dextran increased tau droplet formation in a dose-dependent manner.
  • These tau puncta exhibited liquid-like characteristics initially.
  • Over time, tau droplets transitioned to a more solid-like, aggregated state.

Conclusions:

  • Intracellular accumulation of negatively charged substances or vesicles promotes tau phase separation in vivo.
  • This process contributes to the pathological aggregation of tau protein in tauopathies.
  • Findings suggest potential therapeutic strategies targeting cellular conditions that drive tau aggregation.