Summary
A variant of the Feulgen reaction which has been proposed as a method for demonstrating cytoplasmic DNA in oocytes has been tested on ovarian material from a variety of species. While Schiff positive staining was developed, this was not removable by pretreatment with DNase and could be reproduced by using oxidants used in the pseudoplasmal reaction. This method was not considered useful for demonstrating cytoplasmic DNA.
When chloroform extracted solutions of methyl green were used to stain ovaries, cytoplasmic staining identical in pattern to that obtained with other basic dyes was observed. The cytoplasmic staining was prevented by pretreatment of sections with RNase, but was not affected by DNase pretreatment. In somatic cells with high concentrations of cytoplasmic RNA, only nuclear staining was observed. This nuclear staining was labile to DNase but not to RNase.
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Literature
Brachet, J.: The biochemistry of development. New York: Pergamon Press 1960.
—, and J. Quertier: Cytochemical detection of cytoplasmic deoxyribosenucleic acid (DNA) in amphibian ovocytes. Exp. Cell Res. 32, 410–413 (1963).
Goldstein, D.J.: Mechanism of differential staining of nucleic acids. Nature (Lond.) 191, 407–408 (1961).
Grosch, D.S.: Feulgen positive cytoplasm of Molgula eggs. Nature (Lond.) 181, 1078 (1958).
Kurnick, N.B.: Lack of inhibition of desoxyribonuclease by heat depolymerized desoxyribosenucleic acid. J. Amer. chem. 76, 4040–4041 (1954).
—: Pyronin Y in the methyl-green-pyronin histological stain. Stain Technol. 30, 213–230 (1955).
Nass, M.M.K., S. Nass, and B.A. Afzelius: The general occurrence of mitochondrial DNA. Exp. Cell Res. 37, 516–539 (1965).
Pitelka, D. R.: Observations on the kinetoplast mitochondrion and the cytostome of Bodo. Exp. Cell Res. 25, 87–93 (1961).
Pollister, A.W., and C. Leuchtenberger: The nature of the specificity of methyl green for chromatin. Proc. nat. Acad. Sci. (Wash.) 35, 111–116 (1949).
Rosenkranz, H.S., and A. Bendich: On the nature of the deoxyribonucleic acid-methyl green reaction. J. biophys. biochem. Cytol. 4, 663–664 (1958).
Swift, H.: Cytochemical techniques for nucleic acids. In: The nucleic acids, E. Chargaff and J.N. Davidson, Eds., vol. 2, pp. 51–92. New York: Academic Press.
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This work was supported by U.S. Public Health Service grants GM-10003-03 and K-3-6176-03.
Contribution number 376 of the Bermuda Biological Station.
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Cowden, R.R. The staining behavior of oocyte cytoplasm with the alcoholic Feulgen variant and with methyl green. Histochemie 5, 441–447 (1965). https://doi.org/10.1007/BF00306296
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DOI: https://doi.org/10.1007/BF00306296