Microscopy and Microanalysis



Biological Applications

Altered Mitochondrial Structure and Motion Dynamics in Living Cells with Energy Metabolism Defects Revealed by Real Time Microscope Imaging


Nhu-An  Pham  a1 , Tim  Richardson  a1 , Jessie  Cameron  a2 , Bruno  Chue  a1 and Brian H.  Robinson  a2 c1
a1 Richardson Technologies Inc. at The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada
a2 The Research Institute, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada

Article author query
pham n   [PubMed][Google Scholar] 
richardson t   [PubMed][Google Scholar] 
cameron j   [PubMed][Google Scholar] 
chue b   [PubMed][Google Scholar] 
robinson bh   [PubMed][Google Scholar] 

Abstract

Using the real time microscope (RTM), a system applying new developments in light microscopy, we documented the spatial and temporal dynamics of mitochondrial behavior in human cultured skin fibroblasts. Without the use of stains or probes, we resolved fibroblast mitochondria as dark slender filaments of approximately 0.2 [mu]m wide and up to 10 [mu]m long, as well as a few smaller ovoid forms. In the living cell, the three most common mitochondrial movements were: (1) small oscillatory movements; (2) larger movements including filament extension, retraction, and branching as well as combinations of these actions; and (3) whole transit movements of single mitochondrial filaments. Skin fibroblasts from patients with mitochondrial complex I deficiency and normal fibroblasts during incubation with rotenone, or antimycin A, contained higher proportions of mitochondria in the swollen filamentous forms, nodal filaments, and ovoid forms rather than the slender filamentous forms in normal cells. Interestingly, decreased motility was observed with the more ovoid mitochondrial forms compared to the filamentous forms. We conclude that mitochondrial morphology and dynamic motion are strongly associated with changes in mitochondrial energy metabolism. Images documenting our observations are presented both at single time points and as QuickTime videos. a

(Received November 4 2002)
(Accepted May 6 2003)


Key Words: mitochondria; live; QuickTime; respiratory chain; real-time.

Correspondence:
c1 Corresponding author. E-mail: bhr@sickkids.ca.


Footnotes

a Abbreviations: EM: electron microscope; L/P: lactate/pyruvate; QT: QuickTime; CCCP: carbonyl cyanide 3-chloro-phenylhydrazone; CMXRos, chloromethyl-X-rosamine; Rh123, Rhodamine 123; RTM, real time microscope