Histochemistry

, Volume 70, Issue 1, pp 29–32 | Cite as

A histochemical study of protein-bound SH and SS in chromosomes of hyacinth and fava bean root tips

  • D. A. Wiese
Article
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Summary

Root tips were fixed in ethanol-acetic acid and in neutral formalin containing a mercaptide-forming agent to protect thiols from autoxidation. Serial paraffin sections 2 μm thick were labelled for thiols, disulfides and both together with the azogenic maleimide ‘APM’. Nuclei were stained somewhat lighter for thiols, and nucleoli much darker, than was the surrounding cytoplasm; disulfide staining was paler and more evenly distributed. However, regardless of the fixative employd, mitotic chromosomes (precisely localized by fluorescent Feulgen counterstaining) contained negligible amounts of either thiols or disulfides. Although published biochemical studies also have indicated low concentrations of cyst(e)inyl residues in chromosomal proteins, the present findings contradict most previous histochemical reports.

Keywords

Disulfide Paraffin Thiol Formalin Paraffin Section 
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References

  1. Ackerman G, Sneeringer S (1960) Histochemical localization of sulfhydryl groups in the cell of normal human blood and bone marrow. Lab Invest 9:356–363Google Scholar
  2. Bachmann K, Cowden R (1965) Quantitative cytophotometric studies on polyploid liver cell nuclei of frog and rat. Chromosoma 17:181–193Google Scholar
  3. Barrnett R, Seligman A (1952) Histochemical detection of protein-bound sulfhydryl groups. Science 116:323–327Google Scholar
  4. DeLange R, Smith E (1975) Histone function and evolution as viewed by sequence studies. In: Fitzsimons D, Wolstenholme G (eds) The structure and function of chromatin, Ciba Foundation Symposium 28 (new Series). Associated Scientific Publishers, New York, pp 59–76Google Scholar
  5. Elgin S, Stumph W (1975) Chemistry of the non-histone chromosomal proteins. In: Fitzsimons D, Wolstenholme G (eds) The structure and function of chromatin. Ciba Foundation Symposium 28 (new Series). Associated Scientific Publishers, New York, pp 113–130Google Scholar
  6. Hilton J, Stocken L (1966) The role of thiol groups in the modification of the template activity of histone-deoxyribonucleic acid complexes. Biochem J 100:21cGoogle Scholar
  7. Hyde B (1961) An evaluation of the Barrnett and Seligman procedure when used to determine changes in the ratio of nuclear protein-bound disulfide to sulhydryl groups during mitosis. J Histochem Cytochem 9:640–646Google Scholar
  8. Hyde B, Paliwal R (1959) The Barrnett-Seligman sulfhydryl reaction for plant meristems. Stain Technol 34:175–186Google Scholar
  9. Kawamura N (1960) Cytochemical and quantitative study of protein-bound sulfhydryl and disulfide groups in eggs of Arbacia during the first cleavage. Exp Cell Res 20:127–138Google Scholar
  10. Kawamura N, Dan K (1958) Cytochemical study of protein-bound sulfhydryl groups of sea urchin eggs during the first cleavage. J Biophys Biochem Cytol 4:615–625Google Scholar
  11. Nöhammer G, Schaunstein E, Bajardi F, Unger-Ullman C (1977) Microphotometrical quantification of protein thiols in morphologically intact cells of the cervical epithelium I. Basic results with superficial cells of healthy women. Acta Cytol 21:341–344Google Scholar
  12. Pearse AGE (1968) Histochemistry, vol I, 3rd edn. Churchill Livingstone London, p 728Google Scholar
  13. Ruch F (1966) Determination of DNA content by microfluorometry. In: Wied G (ed) Introduction to quantitative cytochemistry, vol 1 Academic Press, New York, pp 281–294Google Scholar
  14. Sandritter W, Krygier A (1959) Cytophotometrische Bestimmung von proteingebundenen Thiolen in der Mitose und Interphase von HeLa Zellen. Z Krebsforsch 62:595–610Google Scholar
  15. Sippel T (1973) The histochemistry of thiols and disulphides I. The use of N-(4-aminophenyl) maleimide for demonstrating thiol groups. Histochem J 5:413–423Google Scholar
  16. Sippel T (1977) Nonspecific staining in the demonstration of thiols with aryl disulfide reagents. J Histochem Cytochem 25:236Google Scholar
  17. Sippel T (1978) The histochemistry of thiols and disulphides II. Staining patterns in rat tissues. Histochem J 10:597–609Google Scholar
  18. Sivak A, Wolman S (1974) Chromosomal proteins in fixed metaphase cells. Histochemistry 42:345–349Google Scholar
  19. Sumner A (1973) Involvement of protein disulphides and sulfhydryls in chromosome banding. Exp Cell Res 83:438–442Google Scholar
  20. Vallee B, Ulmer D (1972) Biochemical effects of mercury, cadmium and lead. Annu Rev Biochem 41:91–128Google Scholar
  21. Wiese D (1979) Histochemical analysis of nuclear protein thiols and disulfides. Ph. D. dissertation. The University of Michigan, Ann Arbor, MichiganGoogle Scholar

Copyright information

© Springer-Verlag 1980

Authors and Affiliations

  • D. A. Wiese
    • 1
  1. 1.Department of AnatomyThe University of Michigan Medical SchoolAnn ArborUSA

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