Skip to main content
Log in

Mitochondrial biochemical activities and heteroplasmy evolution in established D. subobscura cell line

  • Articles
  • Cell and Tissue Models
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

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.

    Article  PubMed  CAS  Google Scholar 

  • Ashburner, M. Drosophila. Vol. 1. A laboratory handbook. Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 1989.

    Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • DiMauro, S.; Bonilla, E.; Davidson, M.; Hirano, M.; Schon, E. A. Mitochondria in neuromuscular disorders. Biochim. Biophys. Acta. 1366:199–210; 1998.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Google Scholar 

  • Echalier, G.; Ohanessian, A. In vitro culture of Drosophila melanogaster embryonic cells. In Vitro Cell. Dev. Biol. 6A:162–172; 1970.

    Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Hatefi, Y. Preparation and properties of NADH: ubiquinone oxidoreductase (complex I), EC1.6.5.3. Methods Enzymol. 5:11–14; 1978a.

    Google Scholar 

  • Hatefi, Y. Preparation and properties of dihydroubiquinone: cytochrome c oxidoreductase (complex III). Methods Enzymol. 53:35–40; 1978b.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Google Scholar 

  • Shepherd, D.; Garland, P. B. The kinetic properties of citrate synthase from rat liver mitochondria. Biochem J 114:597–610; 1969.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Simon, D. K.; Johns, D. R. Mitochondrial disorders: clinical and genetic features. Annu. Rev. Med. 50:111–127; 1999.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • Wibom, R.; Hultman, E. ATP production rate in mitochondria isolated from microsamples of human muscle. Am. J. Physiol. 259:E204-E209; 1990.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to S. Alziari.

Rights and permissions

Reprints 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

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1290/0601003.1

Key words