Isolation of mitochondria from ejaculated ram spermatozoa

Benjamin Bartoov, Glenda Y. Messer

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

A method for isolating a mitochondrial fraction from ejaculated ram spermatozoa is described. It includes treatment with 10 mM dithiothreitol at 0°C for 20 mm followed by homogenization, fractionation on a sand column, and differential centrifugation. The isolated fraction contained about 0.3% of the total sperm DNA, showed a fivefold enrichment in cytochrome oxidase and succinate dehydrogenase activities, and twofold enrichment of NADH dehydrogenase activity. The mitochondrial fraction comprised mainly residual mitochondrial helices, and individual, crescent-shaped mitochondria measuring 0.165 μm wide and 1.256 μm long.

Original languageEnglish
Pages (from-to)68-76
Number of pages9
JournalJournal of Structural Biology
Volume57
Issue number1
DOIs
StatePublished - Oct 1976

Bibliographical note

Funding Information:
We are grateful to Professor M. Herzberg und Dr. S. Grossman, of Bar-Ilan University, for their cooperation and helpful discussions. We thank Dr. N. Ayalon, Mr. M. Eisenberg, and Mr. Y. Lewis, of the Kimron Veterinary Institute, Belt Dagan, for their assistance during the course of this study. We gratefully acknowledge the contribution of Miss J. Fisher and the excellent technical assistance of Mrs. Z. Goldwitch in chemical analysis, Mrs. V. Harel in electron microscopy and Mr. U. Yoran in photography. This work was supported by the Research Committee of Bar-Ilan University, Research Grant No. 161-83-26-4.

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