Document Detail


A rapid method for the isolation of metabolically active mitochondria from rat neurons and astrocytes in primary culture.
MedLine Citation:
PMID:  9507134     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A rapid method (about 1.5 h) for the isolation of intact functional mitochondria from neurons and astrocytes in primary culture is described. Mitochondria isolated by this method are metabolically active and tightly coupled as shown by respiratory control ratio values, which were about 4 with glutamate-malate as substrate. The activities of marker enzymes revealed the occurrence of a low degree of cytosolic (5%) or synaptosomal (5.5%) contamination in the mitochondrial fractions. In addition, the activity of citrate synthase was increased by 4 fold in both neuronal and astrocytic mitochondria with respect to values found in cell homogenates. These results confirm that the method affords mitochondrial preparations from cultured brain cells at suitable levels of purity and enrichment for the study of their mitochondrial function. Since mitochondrial damage has been associated with the pathogenesis of certain neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases (P. Chagnon, C. Betard, Y. Robitaille, A. Cholette, D. Gauvreau, Distribution of brain cytochrome oxidase activity in various neurodegenerative disease, Neuroreport 6 (1995) 711-715 [6]; S.J. Kish, C. Bergeron, A. Rajput, S. Dozic, F. Mastrogiacomo, L. Chang, J.M. Wilson, L.M. DiStefano, J.N. Nobrega, Brain cytochrome oxidase in Alzheimer's disease, J. Neurochem. 59 (1992) 776-779 [10]; A.H.V. Schapira, J.M. Cooper, D. Dexter, J.B. Clark, P. Jenner, C.D. Marsden, Mitochondrial complex I deficiency in Parkinson's disease, J. Neurochem. 54 (1990) 823-827 [15]), the method described here shed light on the possible susceptibility of neuronal or astrocytic mitochondria to deleterious effects of these diseases.
Authors:
A Almeida; J M Medina
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Brain research. Brain research protocols     Volume:  2     ISSN:  1385-299X     ISO Abbreviation:  Brain Res. Brain Res. Protoc.     Publication Date:  1998 Mar 
Date Detail:
Created Date:  1998-05-05     Completed Date:  1998-05-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9716650     Medline TA:  Brain Res Brain Res Protoc     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  209-14     Citation Subset:  IM    
Copyright Information:
Copyright 1998 Elsevier Science B.V.
Affiliation:
Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Salamanca, Edificio Departamental, Avda. del Campo Charro, 37007 Salamanca, Spain.
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MeSH Terms
Descriptor/Qualifier:
Animals
Astrocytes / ultrastructure*
Biological Markers
Cell Respiration / physiology
Cells, Cultured
Mitochondria / enzymology,  metabolism*
Neurons / ultrastructure*
Rats
Rats, Wistar
Time Factors
Chemical
Reg. No./Substance:
0/Biological Markers

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