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Electron microscopic radioautography of intramitochondrial RNA synthesis of HeLa cells in culture

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HeLa cells were cultivated in vitro and incubated in a medium containing 3H-uridine (20 μc/ml) for 30 minutes 1, 2, 4, 8 and 18 hours, doubly fixed with 2.5% glutaraldehyde and 1% osmium tetroxide, embedded in Epon or hydroxypropyl methacrylate, radioautographed with Sakura NR-H2 emulsion, exposed for 40 days, developed with gold-latensification and elon-ascorbic acid developer.

As the results, silver grains appeared not only in the chromatin, nucleoli, endoplasmic reticulum and ribosomes but also in the mitochondria. Most of the silver grains found in the mitochondria were localized on the mitochondrial matrix, while some others were on the cristae and the mitochondrial membranes. These silver grains were removed with RNase digestion. The percentage of labeled mitochondria increased in accordance with the time of incubation. Almost all the mitochondria were labeled within 18 hours.

From the results, it was concluded that the mitochondria synthesized RNA at their matrices.

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Nagata, T. Electron microscopic radioautography of intramitochondrial RNA synthesis of HeLa cells in culture. Histochemie 32, 163–170 (1972). https://doi.org/10.1007/BF00303731

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