Mitochondria in the flight muscles of insects. II. Effects of the medium on the size form,and organization of isolated sarcosomes |
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| Authors: | WATANABE M I WILLIAMS C M |
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| Affiliation: | From the Pioneering Research Laboratories, United States Army Quartermaster Corps, Philadelphia, and The Biological Laboratories, Harvard University, Cambridge |
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| Abstract: | 1. The sarcosomes of Drosophila and the blowfly, Phormia, are dense, spherical, homogeneous bodies when isolated from flight muscle and promptly examined under the phase contrast, oil immersion objective. 2. Their average diameter in newly emerged flies is about 1 micro. This value increases rapidly during the 1st week of adult life and then becomes constant at approximately 2.5 micro. At each age the variation in sarcosome diameter conforms approximately to a normal distribution. 3. The degree to which isolated sarcosomes retain their initial size and organization is remarkably conditioned by the composition and the hydrogen ion concentration of the medium in which they are teased and suspended. In suboptimal media three major categories of change were encountered: (1) swelling, with or without compaction of the contents (as in distilled water and salt solutions); (2) shrinkage to rod-like, pleomorphic forms (as in blood serum); and (3) fuzzy degeneration (as in sugar solutions). 4. The membrane that surrounds each sarcosome becomes plainly visible in swollen sarcosomes. A continuation of swelling is accompanied by the escape of the sarcosomal contents, the vacated membrane persisting as a spherical, optically empty ghost. 5. Sarcosomes appear to behave like osmometers when suspended in various aqueous solutions. Solutes which penetrate the membrane show only transient effects in preventing the osmotic entry of water. 6. Under this analysis we find the membrane to be more or less freely permeable to the ions of sodium, potassium, calcium, magnesium, chloride, and phosphate, to non-electrolytes smaller than hexoses, to phosphorylated hexoses, and to several intermediates of the citric acid cycle. 7. The sarcosomal membrane appears to be less permeable to non-electrolytes larger than pentoses, provided that such molecules are not phosphorylated. 8. The membrane shows a higher permeability to ATP than to ADP. The significance of this observation is considered with respect to the ADP-ATP shuttle between sarcosomes and muscle fibrils. 9. Simple solutions of electrolytes or non-electrolytes cause more or less conspicuous changes in the microscopic appearance of sarcosomes. Prolonged preservation was achieved only in more complicated media containing protein. It is concluded that the Donnan equilibrium is the source of the principal osmotic forces regulating the movement of water through the sarcosomal membrane. 10. The optimal medium for the preservation of isolated sarcosomes was an intracellular Ringer solution containing 2.5 per cent crystalline bovine albumin in 0.16 M potassium phosphate buffer at pH 7.0. |
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