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Preparation and some respiratory properties of coupled mitochondria from ribbed mussel modiolus demissus gill tissue

, : Preparation and some respiratory properties of coupled mitochondria from ribbed mussel modiolus demissus gill tissue. Journal of Experimental Zoology 229(1): 55-68

Preliminary experiments with 2,4-dinitrophenol, 2,4,6-trinitrophenol and NaN3 indicated that most of the O2 consumption by ribbed mussel gill tissue is the result of mitochondrial respiration. A procedure using isoosmotic sucrose, EGTA [ethylene glycolbis (.beta.-aminoethyl ether) N,N,N',N'-tetraacetic acid] defatted serum albumin and HEPES as the isolation medium was devised for the preparation of fully coupled ribbed mussel gill mitochondria. Optimal rates of respiration and respiratory coupling required substrate, ADP, Pi and a fairly high KCl concentration (90 mM) in the assay medium. Glutamate, proline, malate and succinate stimulated O2 consumption with high respiratory control indices and P/O of 3, 3, 3 and 2, respectively. Pyruvate was a weak stimulator of mitochondrial respiration and showed a low respiratory control index with a low P/O. Preparation of gill mitochondria in isoosmotic solutions containing high KCl concentrations (150 mM) yielded mitochondria showing state 2 respiration, slow partially uncoupled ATP synthesis during state 3 respiration and no state 4 respiration. D-mannitol was not used in the mitochondrial isolation or assay medium because of the probable presence of a D-mannitol oxidase in these gill mitochondria.


DOI: 10.1002/jez.1402290108

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