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Dergiler / Turkish Journal of Medical Sciences / 2013 / Cilt: 43 - Sayı: 5

Determination of mitochondrial fragmentation and autophagosome formation in C2C12 skeletal muscle cells

Saadet TÜRKSEVEN
Antonio ZORZANO
Sayfa
775–781
Yayın Tarihi
2013-05-01
DOI
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Conclusion: The findings of this study will enhance our understanding of the mitophagic process in C2C12 skeletal muscle cells in order to identify molecular mechanisms governing removal of dysfunctional mitochondria.

📌 3 Maddede Bu Makale

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Amaç & Hipotez:To investigate the effect of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) on mitochondrial elimination in C2C12 skeletal muscle cells.
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Temel Bulgular & Veri:Materials and methods: C2C12 muscle cells stably expressing Mito-DsRed were treated with DMSO or CCCP at the doses of 10, 20, and 30 µM for 3, 6, and 24 h.
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Sonuç & Katkı:Conclusion: The findings of this study will enhance our understanding of the mitophagic process in C2C12 skeletal muscle cells in order to identify molecular mechanisms governing removal of dysfunctional mitochondria.
🔬Tür:Araştırma Makalesi
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:Key words: Mitophagy · LC3 · skeletal muscle cell

Özet

To investigate the effect of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) on mitochondrial elimination in C2C12 skeletal muscle cells. Materials and methods: C2C12 muscle cells stably expressing Mito-DsRed were treated with DMSO or CCCP at the doses of 10, 20, and 30 µM for 3, 6, and 24 h. After specific treatments, cells were fixed in 4% paraformaldehyde and immunocytochemistry assays were performed using appropriate antibodies to evaluate mitochondrial morphology and autophagosome formation. Total cell lysates were used for western blot assays. Results: CCCP induces mitochondrial fragmentation in C2C12 cells. In response to a mitochondrial uncoupler, endogenous autophagosome formation is significantly increased (P < 0.001). Co-localization analyses of confocal images indicate that fragmented mitochondria undergo engulfment by autophagosomes. Conclusion: The findings of this study will enhance our understanding of the mitophagic process in C2C12 skeletal muscle cells in order to identify molecular mechanisms governing removal of dysfunctional mitochondria.

Abstract

To investigate the effect of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) on mitochondrial elimination in C2C12 skeletal muscle cells. Materials and methods: C2C12 muscle cells stably expressing Mito-DsRed were treated with DMSO or CCCP at the doses of 10, 20, and 30 µM for 3, 6, and 24 h. After specific treatments, cells were fixed in 4% paraformaldehyde and immunocytochemistry assays were performed using appropriate antibodies to evaluate mitochondrial morphology and autophagosome formation. Total cell lysates were used for western blot assays. Results: CCCP induces mitochondrial fragmentation in C2C12 cells. In response to a mitochondrial uncoupler, endogenous autophagosome formation is significantly increased (P < 0.001). Co-localization analyses of confocal images indicate that fragmented mitochondria undergo engulfment by autophagosomes. Conclusion: The findings of this study will enhance our understanding of the mitophagic process in C2C12 skeletal muscle cells in order to identify molecular mechanisms governing removal of dysfunctional mitochondria.

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Bu Makaleye Atıf Yap

TÜRKSEVEN, S., ZORZANO, A. (2013). Determination of mitochondrial fragmentation and autophagosome formation in C2C12 skeletal muscle cells. Turkish Journal of Medical Sciences, Cilt: 43 - Sayı: 5, 775–781. https://ajindex.com/articles/400876

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