Conference on Biophotons 1999 - PRESERVATION OF THE ABILITY OF ISOLATED MITOCHONDRIA TO STRUCTURE SELF-ORGANIZATION

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Conference on Biophotons 1999
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PRESERVATION OF THE ABILITY OF ISOLATED MITOCHONDRIA TO STRUCTURE SELF-ORGANIZATION

A.V. Temnov, V.I. Popov, T.V. Sirota, I.R. Saakyan, I.G. Stavrovskya,

N.I. Fedotcheva, and M.N. Kondrashova

Institute of Theoretical and Experimental Biophysics RAS, Pushchno, 142292, Russia.

E-mail: kondrashova@venus.iteb.serpukhov. su

Functional interaction and distant action in living tissues is related also to structure organization. The ability to structure self-organization is a property highly sensitive to conditions and is poorly preserved under isolation from the organism. In particular, mitochondria isolated in sucrose solutions in which fundamental investigations of respiration and phosphorylation were carried out dissociate into single granules, i.e. lose their native property to gather into filaments.

When potassiun chloride is used for preparation mitochondria preserve native property to form filamentous associations in homogenate (native mitochondria, Biochemistry, Moscow,1997, 62, 159). In this study the influence of conditions of preparation on structure organization and functions of mitochondria in rat liver homogenate was investigated.

Structures of mitochondria were registered by video-camera in dark field of light microscope (x up to 2000) with the following computer morphometric treatment.

Preparation of homogenate in potassium medium at 15 C instead of accepted 0 C provides improvement of self-organisation during its storage.

When added to incubation media mitochondria in homogenate exhibit well pronounced coupled respiration, potential generation and ion transport. These functions are stable for 5 -6 hours and are well pronounced even next day after isolation.

It was found that better preservation of associations favors to substrate level phosphorylation while dissociation of associations provide activation of oxidative phosphorylation and ion transport.

The three-dimensional associations 5 -10 M up to 50 M in size were observed in homogenate without staining and fixation. Enlargement of associations is observed during storage for an hour in particular under warm preparation and storage. The influence of medium pretreated with negative air ions (superoxide + hydrogen peroxide, Biochemistry, Moscow, 1997, 62, 1089; FEBS Lett.1997, 410, 309) considerably increases size of associations up to 200 M and more. 20 min action of air ions on homogenate reversed damage of ultrastructure after 2.5 hour storage in ice.The great variety of structures were observed in mitochondria stained with nitro-tetrazolium blue for succinate dehydrogenase, among them - classic filaments with beads, formations like algae, palmtree and fern leafs.

Native mitochondria in homogenate serve as a convenient model to study mechanisms of self-organization of living tissues and have advantages (rapid preparation, less than 10 min, minimal amount of tissue, up to 100 mg, stability) for clinical investigations.

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