Related Experiment Video
Updated: Mar 16, 2026
08:35
In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells
Published on: June 12, 2017
10.8K
Muscle LIM protein/CSRP3: a mechanosensor with a role in autophagy
M M Rashid1, A Runci1, L Polletta1
1Department of Experimental Medicine, University of Rome , Sapienza, Rome, Italy.
Cell Death Discovery
|August 24, 2016
Summary
Muscle LIM protein (MLP) regulates muscle cell autophagy and differentiation. MLP silencing impairs autophagosome formation and flux, leading to reduced differentiation and increased cell death.
Area of Science:
- Muscle biology
- Cellular autophagy
- Protein interactions
Background:
- Muscle LIM protein (MLP) is crucial for muscle development and maintenance.
- Muscle cells require autophagy for differentiation and structural integrity.
Purpose of the Study:
- To investigate the role of MLP in muscle cell autophagy.
- To elucidate the mechanism by which MLP influences autophagosome formation and flux.
Main Methods:
- C2C12 mouse myoblasts were used, with MLP silenced or overexpressed.
- Co-immunoprecipitation and proximity ligation assay (PLA) were employed to study MLP/LC3 interaction.
- Myoblast differentiation markers (MyoD1, MyoG1, myosin heavy chain) and autophagic flux were assessed.
- Ultrastructural analysis and apoptosis assays (caspase-3, PARP cleavage) were performed.
Main Results:
- MLP silencing decreased LC3-II levels and impaired long-lived protein degradation, indicating reduced autophagic flux.
- MLP deficiency hindered myoblast differentiation and led to abnormal autophagosome structures.
- MLP-silenced cells exhibited increased susceptibility to apoptotic cell death.
Conclusions:
- MLP is essential for proper autophagosome formation and flux in muscle cells.
- MLP interacts with LC3-II, regulating autophagy during muscle differentiation and maintenance.
- Disruption of MLP function compromises muscle cell integrity and survival.
Related Concept Videos
01:27Autophagy
6.0K
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,...
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
6.0K
01:22PI3K/mTOR/AKT Signaling Pathway
6.1K
The mammalian target of rapamycin (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast, mTORC2 consists of a...
6.1K
01:36Delivery Pathways to the Lysosome
10.4K
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...
Endocytosis
In endocytosis, the cell membrane takes up macromolecules and particles from the surrounding medium. Clathrin-mediated...
10.4K
03:03mTOR Signaling and Cancer Progression
5.0K
The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
The mTOR pathway or the...
5.0K
01:07MAPK Signaling Cascades
9.0K
Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
9.0K
01:18Autophagic Cell Death
4.9K
Christian de Duve discovered “autophagy,” a process in which cellular components are engulfed by membrane-bound organelles called autophagosomes. The autophagosomes then fuse with lysosomes to digest the enclosed contents. Autophagy is generally activated in cells to prevent cell death. However, cell death is triggered when the damage is beyond repair.
Autophagy and Apoptosis
Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and...
Autophagy and Apoptosis
Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and...
4.9K