Abstract
A quantitative ultrastructural radioautographic study of in vitro protein synthesis has been carried out in rat synaptosomal fractions incubated with tritiated leucine or a tritiated amino acid mixture. Analysis of grain density distribution demonstrated that presynaptic endings are labeled. 30–50% of the developed grains, representing tritiated amino acids incorporated into proteins, were related to presynaptic endings which accounted for 75–77% of the total processes. 34–45% of the grains were related to processes containing ribosomes which accounted for only 4–7% of the total processes. The relative specific activity of these ribosome-containing processes, some of which could be identified as postsynaptic elements, was up to ten times higher than that of the presynaptic ending. These findings indicate that protein synthesis takes place in vitro in presynaptic terminals although to a significantly lesser degree than that occurring in ribosome-containing processes, which, with other nonpresynaptic processes, are at the present time unavoidable contaminants of synaptosomal fractions. Presynaptic endings that in radioautographs contained no mitochondria were labeled. Also, presynaptic endings were labeled after incubation in the presence of chloramphenical which inhibited 20% of the protein synthesis of the synaptosomal fraction. It is concluded that besides mitochondrial protein synthesis, another protein synthesizing system operates in presynaptic endings in vitro.
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Selected References
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- Autilio L. A., Appel S. H., Pettis P., Gambetti P. L. Biochemical studies of synapses in vitro. I. Protein synthesis. Biochemistry. 1968 Jul;7(7):2615–2622. doi: 10.1021/bi00847a025. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B., Hemsworth B. A. Intraneural mitochondria. Incorporation of amino acids and monosaccharides into macromolecules by isolated synaptosomes and synaptosomal mitochondria. J Biol Chem. 1970 Jan 25;245(2):363–371. [PubMed] [Google Scholar]
- Droz B., Barondes S. M. Nerve endings: rapid appearance of labeled protein shown by electron microscope radioautography. Science. 1969 Sep 12;165(3898):1131–1133. doi: 10.1126/science.165.3898.1131. [DOI] [PubMed] [Google Scholar]
- Faeder I. R., Salpeter M. M. Glutamate uptake by a stimulated insect nerve muscle preparation. J Cell Biol. 1970 Aug;46(2):300–307. doi: 10.1083/jcb.46.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gonatas N. K., Autilio-Gambetti L., Gambetti P., Shafer B. Morphological and biochemical changes in rat synaptosome fractions during neonatal development. J Cell Biol. 1971 Nov;51(21):484–498. doi: 10.1083/jcb.51.2.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haldar D. Protein synthesis in isolated rat brain mitochondria. Biochem Biophys Res Commun. 1971 Mar 5;42(5):899–904. doi: 10.1016/0006-291x(71)90515-8. [DOI] [PubMed] [Google Scholar]
- Hedley-Whyte E. T., Rawlins F. A., Salpeter M. M., Uzman B. G. Distribution of cholesterol-1,2-H3 during maturation of mouse peripheral nerve. Lab Invest. 1969 Dec;21(6):536–547. [PubMed] [Google Scholar]
- Henson C. P., Weber C. N., Mahler H. R. Formation of yeast mitochondria. I. Kinetics of amino acid incorporation during derepression. Biochemistry. 1968 Dec;7(12):4431–4444. doi: 10.1021/bi00852a040. [DOI] [PubMed] [Google Scholar]
- Kopriwa B M. A semiautomatic instrument for the radioautographic coating technique. J Histochem Cytochem. 1966 Dec;14(12):923–928. doi: 10.1177/14.12.923. [DOI] [PubMed] [Google Scholar]
- Lemkey Johnston N., Larramendi L. M. The separation and identification of fractions of nonmyelinated axons from the cerebellum of the cat. Exp Brain Res. 1968;5(4):326–340. doi: 10.1007/BF00235907. [DOI] [PubMed] [Google Scholar]
- Mahler H. R., Jones L. R., Moore W. J. Mitochondrial contribution to protein synthesis in cerebral cortex. Biochem Biophys Res Commun. 1971 Feb 5;42(3):384–389. doi: 10.1016/0006-291x(71)90382-2. [DOI] [PubMed] [Google Scholar]
- Morgan I. G., Austin L. Synaptosomal protein synthesis in a cell-free system. J Neurochem. 1968 Jan;15(1):41–51. doi: 10.1111/j.1471-4159.1968.tb06172.x. [DOI] [PubMed] [Google Scholar]
- REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salpeter M. M., Bachmann L., Salpeter E. E. Resolution in electron microscope radioautography. J Cell Biol. 1969 Apr;41(1):1–32. doi: 10.1083/jcb.41.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salpeter M. M. H3-proline incorporation into cartilage: electron microscope autoradiographic observations. J Morphol. 1968 Apr;124(4):387–421. doi: 10.1002/jmor.1051240402. [DOI] [PubMed] [Google Scholar]