Summary
A mutant strain of drosophila (D. subobscura) has two types of mitochondrial genomes: a small population (20%) identical to that of the wild strain (15.9 kb) and a predominant population (80%) which has undergone a 5-kb deletion affecting more than 30% of the coding zone. Two cell lines were established from homogenates of embryos from mutant and wild strains. The activities of the respiratory complexes measured in the different cell lines are much lower than in the flies, indicating a glycolytic metabolism. Various modifications of the medium composition did not change this metabolic pathway. The mutant cell line has two types of populations of mitochondrial genomes and the heteroplasmy is equivalent to that measured in the mutant strain. However, the biochemical characteristics differ from those observed in the flies (i.e., the decrease of complex I and III activities), and the various systems of compensation for the consequences of the deletion that are showed in the mutant strain are no longer observed. Furthermore, in contrast with observations made on mutant flies, the heteroplasmy appears unstable in the mutant cell lines: after 60 or so generations, it progressively decreases until it disappears completely. The limited importance of mitochondrial energy metabolism in cells may explain the low impact of the mutation on the established cell line, in contrast to what is seen in the mutant strain.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Alziari, S.; Stepien, G.; Durand, R. In vitro incorporation of (35S)-methionine in mitochondrial proteins of Drosophila melanogaster. Biochem. Biophys. Res. Commun. 99:1–8; 1981.
Ashburner, M. Drosophila. Vol. 1. A laboratory handbook. Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 1989.
Beziat, F.; Morel, F.; Volz-Lingenhol, A.; Saint Paul, N.; Alziari, S. Mitochondrial genome expression in a mutant strain of D. subobscura, an animal model for large scale mtDNA deletion. Nucleic Acids Res. 21:387–392; 1993.
Beziat, F.; Touraille, S.; Debise, R.; Morel, F.; Petit, N.; Lecher, P.; Alziari, S. Biochemical and molecular consequences of massive mitochondrial gene loss in different tissues of a mutant strain of Drosophila subobscura. J. Biol. Chem. 272:22583–22590; 1997.
Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254; 1976.
Debise, R.; Touraille, S.; Durand, R.; Alziari, S. Biochemical consequences of a large deletion in the mitochondrial genome of a Drosophila subobscura strain. Biochem. Biophys. Res. Commun. 196:355–362; 1993.
DiMauro, S.; Bonilla, E.; Davidson, M.; Hirano, M.; Schon, E. A. Mitochondria in neuromuscular disorders. Biochim. Biophys. Acta. 1366:199–210; 1998.
Echalier, G. In vitro established lines of Drosophila cells and applications in physiological genetics. Invertebrate tissue culture: applications in medicine, biology and agriculture. 4th Int. Conf. invertebrate tissue culture, Mont Gabriel/Montréal, Canada, Kurstak, E.; Maramorosch, K., ed. New York: Academic Press; 1976:131–150.
Echalier, G.; Ohanessian, A. In vitro culture of Drosophila melanogaster embryonic cells. In Vitro Cell. Dev. Biol. 6A:162–172; 1970.
Errede, B.; Kamen, M. D.; Hatefi, Y. Preparation and properties of complex IV (ferrocytochrome c: oxygen oxidoreductase EC 1.9.3.1). Methods Enzymol. 53:40–47; 1978.
Farge, G.; Touraille, S.; Le Goff, S.; Petit, N.; Renoux, M.; Morel, F.; Alziari, S. The nuclear genome is involved in heteroplasmy control in a mitochondrial mutant strain of Drosophila subobscura. Eur. J. Biochem. 269:998–1005; 2002.
Gerard, B.; Bourgeron, T.; Chretien, D.; Rotig, A.; Munnich A.; Rustin, P. Uridine preserves the expression of respiratory enzyme deficiencies in cultured fibroblasts. Eur. J. Pediatr. 152:270; 1993.
Hatefi, Y. Preparation and properties of NADH: ubiquinone oxidoreductase (complex I), EC1.6.5.3. Methods Enzymol. 5:11–14; 1978a.
Hatefi, Y. Preparation and properties of dihydroubiquinone: cytochrome c oxidoreductase (complex III). Methods Enzymol. 53:35–40; 1978b.
Lecher, P.; Petit, N.; Beziat, F.; Alziari, S. Localization by ultrastructural in situ hybridization of mitochondrial transcripts in epithelial cells of a Drosophila subobscura deletion mutant. Eur. J. Cell Biol. 71:423–427; 1996.
Le Goff, S.; Lachaume, P.; Touraille, S.; Alziari, S. The nuclear genome of a Drosophila mutant strain increases the frequency of rearranged mitochondrial DNA molecules. Curr. Genet. 40:345–354; 2002.
Morel, F.; Debise, R.; Renoux, M.; Touraille, S.; Ragno, M.; Alziari, S. Biochemical and molecular consequences of ethidium bromide treatment on Drosophila cells. Insect Biochem. Mol. Biol. 29:835–843; 1999.
Petit, N.; Touraille, S.; Debise, R.; Morel, F.; Renoux, M.; Lecher, P.; Alziari S. Developmental changes in heteroplasmy level and mitochondrial gene expression in a Drosophila subobscura mitochondrial deletion mutant. Curr. Genet. 33:330–339; 1998.
Schneider, I.; Blumenthal, A. B. Drosophila cell and tissue culture. In: Ashburner, M.; Wright, T. R. F., ed. The genetic and biology of Drosophila. Vol. 2a. New York: Academic Press; 1978:265–315.
Shepherd, D.; Garland, P. B. The kinetic properties of citrate synthase from rat liver mitochondria. Biochem J 114:597–610; 1969.
Simcox, A. A.; Sobeih, M. M.; Shearn, A. Establishment and characterization of continuous cell lines derived from temperature-sensitive mutants of Drosophila melanogaster. Somatic Cell Mol. Genet. 11:63–70; 1985.
Simon, D. K.; Johns, D. R. Mitochondrial disorders: clinical and genetic features. Annu. Rev. Med. 50:111–127; 1999.
Tang, Y.; Manfredi G.; Hirano, M.; Schon, E. A. Maintenance of human rearranged mitochondrial DNAs in long-term cultured transmitochondrial cell lines. Mol. Biol. Cell. 11:2349–2358; 2000.
Volz-Lingenhöhl, A.; Solignac, M.; Sperlich, D. Stable heteroplasmy for a large-scale deletion in the coding region of Drosophila subobscura mitochondrial DNA. Proc. Natl. Acad. Sci. USA 89:11528–11532; 1992.
Wibom, R.; Hultman, E. ATP production rate in mitochondria isolated from microsamples of human muscle. Am. J. Physiol. 259:E204-E209; 1990.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Morel, F., Renoux, M. & Alziari, S. Mitochondrial biochemical activities and heteroplasmy evolution in established D. subobscura cell line. In Vitro Cell.Dev.Biol.-Animal 42, 201–207 (2006). https://doi.org/10.1290/0601003.1
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1290/0601003.1

