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. 1973 Apr 1;57(1):77–87. doi: 10.1083/jcb.57.1.77

ELECTRON MICROSCOPE AUTORADIOGRAPHIC DETECTION OF SITES OF PROTEIN SYNTHESIS IN THE RABBIT RETINA MÜLLER CELLS

M M Magalhães 1, Antonio Coimbra 1
PMCID: PMC2108949  PMID: 4570792

Abstract

Rabbit retinas were incubated in medium containing 500 µCi of [3H]leucine for 3 min, and transferred to medium without isotope for another 7, 17, 37, 57, and 117 min. Retinal pieces were fixed in paraformaldehyde and osmium tetroxide and embedded in Epon. Thin sections were autoradiographed with Ilford L4 emulsion, and a quantitative study of silver grain distribution per Müller cell portion, and per Müller cell organelle, was carried out. Grain density per unit area was high over the middle cell portion at each incubation interval. Silver grains were numerous over background cytoplasm (which comprised free ribosomes) but their percentage was constant at all times and their relative concentration low. Silver grains were numerous and highly concentrated, at pulse incubation, over the rough endoplasmic reticulum (RER) and then decreased sharply, but this decline coincided with an increase over the Golgi complex, peaking at 20 min. Another peak appeared over the cell periphery at 60 min. These findings suggest the simultaneous synthesis of two types of proteins in Müller cells; structural proteins in background cytoplasm and proteins of secretory type in the RER.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AMES A., 3rd, HASTINGS A. B. Studies on water and electrolytes in nervous tissue. I. Rabbit retina: methods and interpretation of data. J Neurophysiol. 1956 May;19(3):201–212. doi: 10.1152/jn.1956.19.3.201. [DOI] [PubMed] [Google Scholar]
  2. Ashley C. A., Peters T., Jr Electron microscopic radioautographic detection of sites of protein synthesis and migration in liver. J Cell Biol. 1969 Nov;43(2):237–249. doi: 10.1083/jcb.43.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COGAN D. G., KUWABARA T. Tetrazolium studies on the retina. II. Substrate dependent patterns. J Histochem Cytochem. 1959 Sep;7:334–341. doi: 10.1177/7.5.334. [DOI] [PubMed] [Google Scholar]
  4. Castle J. D., Jamieson J. D., Palade G. E. Radioautographic analysis of the secretory process in the parotid acinar cell of the rabbit. J Cell Biol. 1972 May;53(2):290–311. doi: 10.1083/jcb.53.2.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hodson S., Marshall J. Tyrosine incorporation into the rabbit retina. J Cell Biol. 1967 Dec;35(3):722–726. doi: 10.1083/jcb.35.3.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. KUWABARA T., COGAN D. G. Tetrzolium studies on the retina: III. Activity of metabolic intermediates and miscellaneous substrates. J Histochem Cytochem. 1960 May;8:214–224. doi: 10.1177/8.3.214. [DOI] [PubMed] [Google Scholar]
  7. Kopriwa B. M. The influence of development on the number and appearance of silver grains in electron microscopic radioautography. J Histochem Cytochem. 1967 Sep;15(9):501–515. doi: 10.1177/15.9.501. [DOI] [PubMed] [Google Scholar]
  8. Magalhàes M. M., Coimbra A. Electron microscope radioautographic study of glycogen synthesis in the rabbit retina. J Cell Biol. 1970 Oct;47(1):263–275. doi: 10.1083/jcb.47.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Magalhães M. M., Coimbra A. The rabbit retina Müller cell. A fine structural and cytochemical study. J Ultrastruct Res. 1972 May;39(3):310–326. doi: 10.1016/s0022-5320(72)90026-3. [DOI] [PubMed] [Google Scholar]
  10. Packman P. M., Blomstrand C., Hamberger A. Disc electrophoretic separation of proteins in neuronal, glial and subcellular fractions from cerebral cortex. J Neurochem. 1971 Mar;18(3):479–487. doi: 10.1111/j.1471-4159.1971.tb11975.x. [DOI] [PubMed] [Google Scholar]
  11. Peters T., Jr, Ashley C. A. An artefact in radioautography due to binding of free amino acids to tissues by fixatives. J Cell Biol. 1967 Apr;33(1):53–60. doi: 10.1083/jcb.33.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Whur P., Herscovics A., Leblond C. P. Radioautographic visualization of the incorporation of galactose-3H and mannose-3H by rat thyroids in vitro in relation to the stages of thyroglobulin synthesis. J Cell Biol. 1969 Nov;43(2):289–311. doi: 10.1083/jcb.43.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Zagury D., Uhr J. W., Jamieson J. D., Palade G. E. Immunoglobulin synthesis and secretion. II. Radioautographic studies of sites of addition of carbohydrate moieties and intracellular transport. J Cell Biol. 1970 Jul;46(1):52–63. doi: 10.1083/jcb.46.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]

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