Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP

Gilad Twig, Solomon A. Graf, Jakob D. Wikstrom, Hibo Mohamed, Sarah E. Haigh, Alvaro Elorza, Motti Deutsch, Naomi Zurgil, Nicole Reynolds, Orian S. Shirihai

Resultado de la investigación: Article

87 Citas (Scopus)

Resumen

Assembly of mitochondria into networks supports fuel metabolism and calcium transport and is involved in the cellular response to apoptotic stimuli. A mitochondrial network is defined as a continuous matrix lumen whose boundaries limit molecular diffusion. Observation of individual networks has proven challenging in live cells that possess dense populations of mitochondria. Investigation into the electrical and morphological properties of mitochondrial networks has therefore not yielded consistent conclusions. In this study we used matrix-targeted, photoactivatable green fluorescent protein to tag single mitochondrial networks. This approach, coupled with real-time monitoring of mitochondrial membrane potential, permitted the examination of matrix lumen continuity and fusion and fission events over time. We found that adjacent and intertwined mitochondrial structures often represent a collection of distinct networks. We additionally found that all areas of a single network are invariably equipotential, suggesting that a heterogeneous pattern of membrane potential within a cell's mitochondria represents differences between discrete networks. Interestingly, fission events frequently occurred without any gross morphological changes and particularly without fragmentation. These events, which are invisible under standard confocal microscopy, redefine the mitochondrial network boundaries and result in electrically disconnected daughter units.

Idioma originalEnglish
PublicaciónAmerican Journal of Physiology - Cell Physiology
Volumen291
N.º1
DOI
EstadoPublished - 2006

Huella dactilar

Mitochondria
Membranes
Confocal microscopy
Mitochondrial Membrane Potential
Green Fluorescent Proteins
Metabolism
Confocal Microscopy
Membrane Potentials
Fusion reactions
Observation
Calcium
Monitoring
Population

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Citar esto

Twig, Gilad ; Graf, Solomon A. ; Wikstrom, Jakob D. ; Mohamed, Hibo ; Haigh, Sarah E. ; Elorza, Alvaro ; Deutsch, Motti ; Zurgil, Naomi ; Reynolds, Nicole ; Shirihai, Orian S. / Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP. En: American Journal of Physiology - Cell Physiology. 2006 ; Vol. 291, N.º 1.
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title = "Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP",
abstract = "Assembly of mitochondria into networks supports fuel metabolism and calcium transport and is involved in the cellular response to apoptotic stimuli. A mitochondrial network is defined as a continuous matrix lumen whose boundaries limit molecular diffusion. Observation of individual networks has proven challenging in live cells that possess dense populations of mitochondria. Investigation into the electrical and morphological properties of mitochondrial networks has therefore not yielded consistent conclusions. In this study we used matrix-targeted, photoactivatable green fluorescent protein to tag single mitochondrial networks. This approach, coupled with real-time monitoring of mitochondrial membrane potential, permitted the examination of matrix lumen continuity and fusion and fission events over time. We found that adjacent and intertwined mitochondrial structures often represent a collection of distinct networks. We additionally found that all areas of a single network are invariably equipotential, suggesting that a heterogeneous pattern of membrane potential within a cell's mitochondria represents differences between discrete networks. Interestingly, fission events frequently occurred without any gross morphological changes and particularly without fragmentation. These events, which are invisible under standard confocal microscopy, redefine the mitochondrial network boundaries and result in electrically disconnected daughter units.",
keywords = "Fission, Fusion, Green fluorescent protein, Heterogeneity, Membrane potential, Tetramethylrhodamine ethyl ester perchlorate",
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Twig, G, Graf, SA, Wikstrom, JD, Mohamed, H, Haigh, SE, Elorza, A, Deutsch, M, Zurgil, N, Reynolds, N & Shirihai, OS 2006, 'Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP', American Journal of Physiology - Cell Physiology, vol. 291, n.º 1. https://doi.org/10.1152/ajpcell.00348.2005

Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP. / Twig, Gilad; Graf, Solomon A.; Wikstrom, Jakob D.; Mohamed, Hibo; Haigh, Sarah E.; Elorza, Alvaro; Deutsch, Motti; Zurgil, Naomi; Reynolds, Nicole; Shirihai, Orian S.

En: American Journal of Physiology - Cell Physiology, Vol. 291, N.º 1, 2006.

Resultado de la investigación: Article

TY - JOUR

T1 - Tagging and tracking individual networks within a complex mitochondrial web with photoactivatable GFP

AU - Twig, Gilad

AU - Graf, Solomon A.

AU - Wikstrom, Jakob D.

AU - Mohamed, Hibo

AU - Haigh, Sarah E.

AU - Elorza, Alvaro

AU - Deutsch, Motti

AU - Zurgil, Naomi

AU - Reynolds, Nicole

AU - Shirihai, Orian S.

PY - 2006

Y1 - 2006

N2 - Assembly of mitochondria into networks supports fuel metabolism and calcium transport and is involved in the cellular response to apoptotic stimuli. A mitochondrial network is defined as a continuous matrix lumen whose boundaries limit molecular diffusion. Observation of individual networks has proven challenging in live cells that possess dense populations of mitochondria. Investigation into the electrical and morphological properties of mitochondrial networks has therefore not yielded consistent conclusions. In this study we used matrix-targeted, photoactivatable green fluorescent protein to tag single mitochondrial networks. This approach, coupled with real-time monitoring of mitochondrial membrane potential, permitted the examination of matrix lumen continuity and fusion and fission events over time. We found that adjacent and intertwined mitochondrial structures often represent a collection of distinct networks. We additionally found that all areas of a single network are invariably equipotential, suggesting that a heterogeneous pattern of membrane potential within a cell's mitochondria represents differences between discrete networks. Interestingly, fission events frequently occurred without any gross morphological changes and particularly without fragmentation. These events, which are invisible under standard confocal microscopy, redefine the mitochondrial network boundaries and result in electrically disconnected daughter units.

AB - Assembly of mitochondria into networks supports fuel metabolism and calcium transport and is involved in the cellular response to apoptotic stimuli. A mitochondrial network is defined as a continuous matrix lumen whose boundaries limit molecular diffusion. Observation of individual networks has proven challenging in live cells that possess dense populations of mitochondria. Investigation into the electrical and morphological properties of mitochondrial networks has therefore not yielded consistent conclusions. In this study we used matrix-targeted, photoactivatable green fluorescent protein to tag single mitochondrial networks. This approach, coupled with real-time monitoring of mitochondrial membrane potential, permitted the examination of matrix lumen continuity and fusion and fission events over time. We found that adjacent and intertwined mitochondrial structures often represent a collection of distinct networks. We additionally found that all areas of a single network are invariably equipotential, suggesting that a heterogeneous pattern of membrane potential within a cell's mitochondria represents differences between discrete networks. Interestingly, fission events frequently occurred without any gross morphological changes and particularly without fragmentation. These events, which are invisible under standard confocal microscopy, redefine the mitochondrial network boundaries and result in electrically disconnected daughter units.

KW - Fission

KW - Fusion

KW - Green fluorescent protein

KW - Heterogeneity

KW - Membrane potential

KW - Tetramethylrhodamine ethyl ester perchlorate

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JO - American Journal of Physiology - Cell Physiology

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