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Related Concept Videos

Autophagy01:27

Autophagy

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Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
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Renewal of Intestinal Stem Cells01:23

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The intestinal epithelial lining rapidly renews every 4 to 5 days. The renewal is facilitated by intestinal stem cells (ISCs) located at the base of the crypt– a gland located at the bottom of each villus. ISCs divide asymmetrically to form new stem cells and progenitor daughter cells. The daughter cells are called transit-amplifying (TA) cells which move upwards along the crypt and either differentiate into absorptive cells– the enterocytes or secretory cells– including the...
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Delivery Pathways to the Lysosome01:36

Delivery Pathways to the Lysosome

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Eukaryotic cells use different mechanisms to eliminate toxic waste obsolete and worn-out substances. Lysosomes play a pivotal role in this, and hence, these substances are carried to the lysosome from other parts of the cell and extracellular space through different pathways. The most elaborately studied pathways to the lysosome are the endocytic pathways.
Endocytosis
In endocytosis, the cell membrane takes up macromolecules and particles from the surrounding medium. Clathrin-mediated...
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Role Of Notch Signalling In Intestinal Stem Cell Renewal01:12

Role Of Notch Signalling In Intestinal Stem Cell Renewal

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Notch signaling was first discovered in Drosophila melanogaster, where it is involved in cell lineage differentiation. Notch signaling regulates the maintenance and differentiation of intestinal stem cells or ISCs by controlling the expression of atonal homolog 1 or Atoh1. Atoh1 directs cells to differentiate into secretory cells.
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
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Related Experiment Video

Updated: Dec 14, 2025

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells
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In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Published on: June 12, 2017

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Autophagy Detection in Intestinal Stem Cells.

Jumpei Asano1,2, Taku Sato1, Toshiaki Ohteki3

  • 1Department of Biodefense Research, Medical Research Institute, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|July 25, 2020
PubMed
Summary

This study details methods to detect autophagy and mitophagy in intestinal stem cells (ISCs). These techniques are crucial for understanding stem cell homeostasis and related diseases.

Keywords:
Atg5AutophagosomeAutophagyGFP-LC3Intestinal epithelial cells (IECs)Intestinal stem cells (ISCs)Lgr5MitochondriaReactive oxygen species (ROS)p62

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Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry

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Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond
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In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells
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Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
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Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond
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Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

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Area of Science:

  • Cell Biology
  • Gastroenterology
  • Stem Cell Biology

Background:

  • Autophagy is a vital lysosomal degradation pathway involved in maintaining cellular balance and disease progression.
  • Intrinsic autophagy within intestinal stem cells (ISCs) plays a critical role in their homeostasis.

Purpose of the Study:

  • To present detailed methodologies for assessing autophagy in ISCs.
  • To describe techniques for evaluating mitophagy within ISCs.

Main Methods:

  • Autophagosome detection in GFP-LC3 transgenic mice.
  • Quantification of p62 protein levels.
  • Assessment of mitochondrial transmembrane potential using MitoTracker.
  • Measurement of reactive oxygen species (ROS) levels using CellROX solution.

Main Results:

  • Established protocols for visualizing and quantifying autophagy in ISCs.
  • Developed methods for assessing mitophagy, including mitochondrial health and oxidative stress markers.

Conclusions:

  • The described methods provide a robust framework for studying autophagy and mitophagy in ISCs.
  • These techniques are essential for future research into ISC function, homeostasis, and disease.