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. 1970 May 1;45(2):235–245. doi: 10.1083/jcb.45.2.235

ELECTRON MICROSCOPY OF THE AMMONIACAL SILVER REACTION FOR HISTONES IN THE ERYTHROPOIETIC CELLS OF THE CHICK

Edith K MacRae 1, Gerald D Meetz 1
PMCID: PMC2107907  PMID: 4105111

Abstract

The product of the postformalin ammoniacal silver reaction, which has been claimed to distinguish lysine-rich from arginine-rich histones with the light microscope on the basis of a color difference, was examined in developing erythroid cells of chick bone marrow with the electron microscope. Stem cells and early erythroblasts exhibit no, or little, ammoniacal silver reaction product, while small basophilic erythroblasts, polychromatophilic erythrocytes, and reticulocytes exhibit an increasing amount of reaction product as maturation proceeds. The reaction product is in the form of discrete electron-opaque particles associated with heterochromatin. The ammoniacal silver reaction in the erythroid cell series is interpreted as reflecting either the accumulation of newly synthesized arginine-rich histones or changes in the availability of reactive sites in preformed histones.

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Selected References

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  1. ALLFREY V. G., FAULKNER R., MIRSKY A. E. ACETYLATION AND METHYLATION OF HISTONES AND THEIR POSSIBLE ROLE IN THE REGULATION OF RNA SYNTHESIS. Proc Natl Acad Sci U S A. 1964 May;51:786–794. doi: 10.1073/pnas.51.5.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ALLFREY V. G., LITTAU V. C., MIRSKY A. E. On the role of of histones in regulation ribonucleic acid synthesis in the cell nucleus. Proc Natl Acad Sci U S A. 1963 Mar 15;49:414–421. doi: 10.1073/pnas.49.3.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. I. Histochemical staining of bone marrow smears. J Cell Biol. 1968 Nov;39(2):286–298. doi: 10.1083/jcb.39.2.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Black M. M., Ansley H. R. Histone specificity revealed by ammoniacal silver staining. J Histochem Cytochem. 1966 Feb;14(2):177–181. doi: 10.1177/14.2.177. [DOI] [PubMed] [Google Scholar]
  5. CAMERON I. L., PRESCOTT D. M. RNA and protein metabolism in the maturation of the nucleated chicken erythrocyte. Exp Cell Res. 1963 May;30:609–612. doi: 10.1016/0014-4827(63)90344-6. [DOI] [PubMed] [Google Scholar]
  6. COHN P., SIMSON P. BASIC AND OTHER PROTEINS IN MICROSOMES OF RAT LIVER. Biochem J. 1963 Aug;88:206–212. doi: 10.1042/bj0880206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Comings D. E. Histones of genetically active and inactive chromatin. J Cell Biol. 1967 Dec;35(3):699–708. doi: 10.1083/jcb.35.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. FRENSTER J. H., ALLFREY V. G., MIRSKY A. E. REPRESSED AND ACTIVE CHROMATIN ISOLATED FROM INTERPHASE LYMPHOCYTES. Proc Natl Acad Sci U S A. 1963 Dec;50:1026–1032. doi: 10.1073/pnas.50.6.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fowler D. J., Goodnight C. J. The cyclic production of 5-hydroxytryptamine in the opilionid. Am Zool. 1966 May;6(2):187–193. doi: 10.1093/icb/6.2.187. [DOI] [PubMed] [Google Scholar]
  10. Freedman M. L., Honig G. R., Rabinovitz M. The role of newly synthesized RNA on nuclear histone synthesis by chicken immature erythrocytes. Exp Cell Res. 1966 Nov-Dec;44(2):263–272. doi: 10.1016/0014-4827(66)90431-9. [DOI] [PubMed] [Google Scholar]
  11. GRASSO J. A., WOODARD J. W., SWIFT H. Cytochemical studies of nucleic acids and proteins in erythrocytic development. Proc Natl Acad Sci U S A. 1963 Jul;50:134–140. doi: 10.1073/pnas.50.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gutierrez R. M., Hnilica L. S. Tissue specificity of histone phosphorylation. Science. 1967 Sep 15;157(3794):1324–1325. doi: 10.1126/science.157.3794.1324. [DOI] [PubMed] [Google Scholar]
  13. HUANG R. C., BONNER J. Histone, a suppressor of chromosomal RNA synthesis. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1216–1222. doi: 10.1073/pnas.48.7.1216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Himes M. An analysis of heterochromatin in maize root tips. J Cell Biol. 1967 Oct;35(1):175–181. doi: 10.1083/jcb.35.1.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kleinsmith L. J., Allfrey V. G., Mirsky A. E. Phosphoprotein metabolism in isolated lymphocyte nuclei. Proc Natl Acad Sci U S A. 1966 May;55(5):1182–1189. doi: 10.1073/pnas.55.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Meetz G. D., MacRae E. K. Ammoniacal silver staining of te erythropoietic cells of the chick. J Histochem Cytochem. 1968 Feb;16(2):148–150. doi: 10.1177/16.2.148. [DOI] [PubMed] [Google Scholar]
  17. Murray K., Vidali G., Neelin J. M. The stepwise removal of histones from chicken erythrocyte nucleoprotein. Biochem J. 1968 Mar;107(2):207–215. doi: 10.1042/bj1070207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pogo B. G., Allfrey V. G., Mirsky A. E. RNA synthesis and histone acetylation during the course of gene activation in lymphocytes. Proc Natl Acad Sci U S A. 1966 Apr;55(4):805–812. doi: 10.1073/pnas.55.4.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pogo B. G., Pogo A. O., Allfrey V. G., Mirsky A. E. Changing patterns of histone acetylation and RNA synthesis in regeneration of the liver. Proc Natl Acad Sci U S A. 1968 Apr;59(4):1337–1344. doi: 10.1073/pnas.59.4.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Robbins E., Borun T. W. The cytoplasmic synthesis of histones in hela cells and its temporal relationship to DNA replication. Proc Natl Acad Sci U S A. 1967 Feb;57(2):409–416. doi: 10.1073/pnas.57.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. STEDMAN E. Cell specificity of histones. Nature. 1950 Nov 4;166(4227):780–781. doi: 10.1038/166780a0. [DOI] [PubMed] [Google Scholar]
  22. Scherrer K., Marcaud L., Zajdela F., London I. M., Gros F. Patterns of RNA metabolism in a differentiated cell: a rapidly labeled, unstable 60S RNA with messenger properties in duck erythroblasts. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1571–1578. doi: 10.1073/pnas.56.5.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seegers W., Janoff A. Mediators of inflammation in leukocyte lysosomes. VI. Partial purification and characterization of a mast cell-rupturing component. J Exp Med. 1966 Nov 1;124(5):833–849. doi: 10.1084/jem.124.5.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wallington E. A. The explosive properties of ammoniacal-silver solutions. J Med Lab Technol. 1965 Oct;22(4):220–223. [PubMed] [Google Scholar]
  25. Zeya H. I., Spitznagel J. K. Cationic protein-bearing granules of polymorphonuclear leukocytes: separation from enzyme-rich granules. Science. 1969 Mar 7;163(3871):1069–1071. doi: 10.1126/science.163.3871.1069. [DOI] [PubMed] [Google Scholar]

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