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The Journal of Cell Biology, Vol 41, 547-561, Copyright © 1969 by Rockefeller University Press

ARTICLE

EFFECTS OF ACRIFLAVINE ON THE MITOCHONDRIA AND KINETOPLAST OF CRITHIDIA FASCICULATA

: Correlation of Fine Structure Changes with Decreased Mitochondrial Enzyme Activity



George C. Hill 1 and Winston A. Anderson 1

1 From the Department of Biochemistry, University of Kentucky Medical School, Lexington, Kentucky 40506, and the Laboratoire de Biologie Cellulaire 4, Université de Paris, Faculté des Sciences, 91-Orsay, France.

Dr. Anderson's present address is the Department of Anatomy, Harvard Medical School, Boston, Massachusetts 02115

The effects of acriflavine on the fine structure and function of the mitochondria and the kinetoplast in Crithidia fasciculata have been investigated. A mitochondrial fraction was prepared by differential centrifugation of cells broken by grinding with neutral alumina. Isolated mitochondria or intact cells revealed by spectrophotometric measurements the presence of cytochromes a + a3, b, c555 and o. After cells were grown in acriflavine for 3–4 days, the fine structure of the mitochondria and their cytochrome content were affected. Cells grown in 5.0 µM acriflavine had a threefold decrease in cytochrome a + a3 and decreased respiratory activity. The mitochondrial preparation from these cells had a fivefold decrease in cytochrome a + a3 and a less but significant decrease of other cytochromes present. There was also a decrease in the mitochondrial enzyme activities of NADH, succinic and L-alpha-glycerophosphate oxidases, and succinic and L-alpha-glycerophosphate dehydrogenases. Dyskinetoplastic cells could be demonstrated after growth in 1.0 µM acriflavine. At 5 µM, 80–90% of the cells were dyskinetoplastic. The kinetoplastic DNA was condensed, nonfibrillar, and did not incorporate thymidine-3H. The mitochondria in these cells had few cristae and were shorter and more swollen than the controls. Acriflavine may induce the fine structure effects we have observed and may affect the formation of the mitochondria in C. fasciculata.

Submitted on October 29, 1968
Revised on December 11, 1968


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