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Observations on the penetration and uptake of trypan blue in embryonic membranes of the mouse

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Summary

Pregnant female Swiss albino mice were injected with 0.4 ml of 1% solution of trypan blue on day 7 1/2, 8 1/2, 10 1/2 and 11 1/2 of gestation. Animals were sacrificed 1–6, 24, 48, 72 and 120 hours after injection and the localization of trypan blue granules was determined in histological sections.

Five hours after injection, the dye began to accumulate in the proximal yolk sac epithelium, and within 24 and 48 hours this cell layer was heavily loaded with dye. The dye never became detectable in any other membranes or the embryo proper. This pattern of dye distribution was the same whether injection was initiated at the 7 1/2 8 1/2 day stage of gestation or was delayed until the 10 1/2–11 1/2 day stage.

Histochemical tests for acid phosphatase and monoamine oxidase revealed that the cells of the vitelline membrane from embryos of trypan blue-injected mothers stained very much less intensely than the vitelline membrane tissue from saline-injected controls.

Implications of these observations for theories of action of trypan blue as a teratogenic agent were discussed.

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References

  • Battle, H. I., and H. W. Laale: Trypan blue induced anomalies in embryos of the zebrafish (Brachydanio rerio). Anat. Rec. 138, 333 (1960).

    Google Scholar 

  • Beaudoin, A. R., and J. G. Wilson: Teratogenic effect of trypan blue on the developing chick. Proc. Soc. exp. Biol. (N. Y.) 97, 85–90 (1958).

    Google Scholar 

  • Beck, F., B. Spence, and J. S. Baxter: Effect of trypan blue on rat embryos. Nature (Lond.) 187, 605–607 (1960).

    Google Scholar 

  • Ferm, V. H.: Permeability of the rabbit blastocyst to trypan blue. Anat. Rec. 125, 745–760 (1956).

    PubMed  Google Scholar 

  • —: Teratogenic effects of trypan blue on hamster embryos. J. Embryol. exp. Morph. 6, 284–287 (1958).

    PubMed  Google Scholar 

  • Fox, M. H., and C. M. Gross: Syndrome of cardiac aberrations associated with malformation of the primitive cardiac loop. Anat. Rec. 121, 294–295 (1955).

    Google Scholar 

  • —, —: Experimental production of a syndrome of congenital cardiovascular defects in rats. Anat. Rec. 124, 189–207 (1956).

    PubMed  Google Scholar 

  • — —: Experimentally produced malformations of the heart and great vessels in rat fetuses. Atrial and caval abnormalities. Anat. Rec. 129, 309–331 (1957).

    PubMed  Google Scholar 

  • Gillman, J. C., C. Gilbert, T. Gillman, and I. Spence: A preliminary report on hydrocephalus, spina bifida and other congenital anomalies in the rat produced by trypan blue. S. Afr. J. med. Sci. 13, 47–90 (1948).

    Google Scholar 

  • — —, I. Spence, and T. Gillman: A further report on congenital anomalies in the rat produced by trypan blue. S. Afr. J. med. Sci. 16, 125–135 (1951).

    Google Scholar 

  • Glenner, G. G., H. J. Burtner, and G. W. Brown: The histochemical demonstration of monoamine oxidase activity by tetrazolium salts. J. Histochem. Cytochem. 5, 591–600 (1957).

    PubMed  Google Scholar 

  • Gomori, G.: Microtechnical demonstration of phosphatase in tissue sections. Proc. Soc. exp. Biol. (N. Y.) 42, 23–26 (1939).

    Google Scholar 

  • —: An improved histochemical technique for acid phosphatase. Stain Technol. 25, 81–85 (1950).

    Google Scholar 

  • —: Microscopic histochemistry, p. 189–194, Chicago: Chicago University Press 1952.

    Google Scholar 

  • Gunberg, D. L.: Spina bifida and the Arnold-Chiari malformation in the progeny of trypan blue injected rats. Anat. Rec. 126, 343–359 (1956).

    PubMed  Google Scholar 

  • Hamburgh, M.: Malformations in mouse embryos induced by trypan blue. Nature (Lond.) 169, 27 (1952).

    Google Scholar 

  • —: The embryology of trypan blue induced abnormalities in mice. Anat. Rec. 119, 409–427 (1954).

    PubMed  Google Scholar 

  • —, and V. Callahan: Differences in response to the teratogenic action of trypan blue in two inbred strains of mice. (In press.)

  • Harm, H.: Der Einfluß von Trypanblau auf Nachkommenschaft trächtiger Kaninchen. Z. Naturforsch. 9, 536–540 (1954).

    Google Scholar 

  • Hoar, R. M., and A. J. Salem: Time of teratogenic action of trypan blue in guinea pigs. Anat. Rec. 141, 173–182 (1961).

    PubMed  Google Scholar 

  • Johnson, E. M., and K. P. Chepnik: Effects of trypan blue on the uptake of tritiated uridine by embryonic rats and their associated membranes. Proc. 6th Ann. Meeting Amer. Terat. Soc., Corpus Christi, Texas 1966.

  • Kalter, H., and J. Warkany: Experimental production of congenital malformations in mammals by metabolic prodecure. Physiol. Rev. 39, 69–115 (1959).

    PubMed  Google Scholar 

  • Kreshover, S. J., H. T. Knighton, and J. A. Hancock jr.: Influence of systematically administered trypan blue on prenatal development of rats and mice. J. dent. Res. 36, 677–683 (1957).

    PubMed  Google Scholar 

  • Langman, L., and H. van Drunen: The effect of trypan blue upon maternal protein metabolism and embryonic development. Anat. Rec. 133, 513–526 (1959).

    PubMed  Google Scholar 

  • Murakami, U., and Y. Kameyama: Manifestation of abnormalities in abnormal intrauterine environment. III. Maldevelopment of the central nervous system, eye, and manifestations of heart abnormalities. Jap. J. Genet. 30, 51–61 (1955).

    Google Scholar 

  • Novikoff, A.B.: Lysosomes and related particles. In: The Cell, ed. by J. Brachet and A. E. Mirsky, vol. II, p. 423–448. New York: Academic Press 1961.

    Google Scholar 

  • Schmidzu, Y.: On the permeability to dyestuffs of the placenta of the albino rat and the white mouse. Amer. J. Physiol. 62, 202–224 (1922).

    Google Scholar 

  • Snell, G. D.: The early embryology of the mouse. In: Biology of the mouse, ed. by. G. D. Snell. chap. 1, p. 1–54. New York: Dover Publ., Inc. 1956.

    Google Scholar 

  • Waddington, C. H., and T. C. Carter: A note on abnormalities induced in mouse embryos by trypan blue. J. Embryol. exp. Morph. 1, 167–179 (1953).

    Google Scholar 

  • —, and M. M. Perry: Teratogenic effects of trypan blue on amphibian embryos. J. Embryol. exp. Morph. 4, 110–118 (1956).

    Google Scholar 

  • Warkany, J., J. G. Wilson, and J. F. Geiger: Myeloschisis and myelomeningocele produced experimentally in the rat. J. comp. Neurol. 109, 35–54 (1958).

    PubMed  Google Scholar 

  • Wilson, J. G.: Teratogenic activity of several azo dyes chemically related to trypan blue. Anat. Rec. 123, 313–326 (1955).

    PubMed  Google Scholar 

  • —, A. R. Beaudoin, and H. J. Free: Studies on the mechanism of the teratogenic action of trypan blue. Anat. Rec. 133, 115–128 (1959).

    PubMed  Google Scholar 

  • Wislocki, G. B.: Experimental studies on fetal absorption. Contr. Embryol. Carneg. Instn. 11, 47–59 (1920).

    Google Scholar 

  • —, H. W. Deane, and E. W. Dempsey: The histochemistry of the rodent placenta. Amer. J. Anat. 78, 281–321 (1946).

    Google Scholar 

  • Woollam, D. H. M., and J. W. Millen: The experimental production of congenital malformations. Roy. Soc. Hlth J. 81, 165–172 (1961).

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy, Albert Einstein College of Medicine, New York, N. Y., USA

    Laslo Nebel & Max Hamburgh

  2. Department of Biology, The City College, New York, N. Y., USA

    Laslo Nebel & Max Hamburgh

Authors
  1. Laslo Nebel
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  2. Max Hamburgh
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Additional information

Dedicated to Professor Berta V. Scharrer on her 60th birthday in friendship and gratitude.

This study was supported by USPHS Research Grant NB 01716 from the National Institute for Neurological Diseases and Blindness, National Institutes of Health and by Grant NIH ST 1 GM 102

This work was carried out in the laboratory of Dr. Max Hamburgh, Department of Anatomy, Albert Einstein College of Medicine, while Dr. Laslo Nebel was a visiting professor on sabbatical leave from the Hebrew University, Hadassah Medical School, Jerusalem, Israel.

Acknowledgment is made to Mr. Dan DiDomizio, who collaborated as a technical assistant in this project, in partial fulfillment for research credit as an undergraduate student in the Department of Biology of the City College of New York.

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Nebel, L., Hamburgh, M. Observations on the penetration and uptake of trypan blue in embryonic membranes of the mouse. Zeitschrift für Zellforschung 75, 129–137 (1966). https://doi.org/10.1007/BF00407150

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  • DOI: https://doi.org/10.1007/BF00407150

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