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Histochemistry

, Volume 81, Issue 4, pp 409–415 | Cite as

Comparative enzyme histochemical study on the visceral yolk sac endoderm in the rat in vivo and in vitro

  • A. Miki
  • P. Kugler
Article

Summary

Histochemical study of the visceral yolk-sac endoderm of the rat was performed in vitro (whole-embryo culture for 24, 48 and 72 h explanted at 9.5 days of gestation) and in vivo (10.5, 11.5 and 12.5 days of gestation) in order to compare the distribution and activity of various enzymes involved in the digestion and energy metabolism in both systems. It was shown that, both in vitro and in vivo γ-glytamyltransferase and dipeptidylpeptidase IV are demonstrable in the apical cell membranes (membranebound hydrolases), while acid phosphatase, dipeptidylpeptidases I, II and acid β-galactosidase are concentrated in the supranuclear vacuoles (lysosomal hydrolases), and cytoplasmic lactate dehydrogenase and mitochondrial enzymes (succinate dehydrogenase, NAD-dependent isocitrate dehydrogenase, cytochrom oxidase) are localized in the whole cytoplasm and mainly in the apical cytoplasm, respectively, of the visceral yolk-sac epithelium. In vivo, the activity of all enzymes increased until 12.5 days, but in vitro, this activity increased only until 48 h after the start of culture (corresponding to 11.5 days in vivo). Comparison of the yolk sacs at 10.5 and 11.5 days in vivo with those after 24 and 48 h in vitro showed that the activities of all the investigated enzymes were almost identical. Yolk sacs which were cultured for 72 h showed lower activities of lysosomal and mitochondrial enzymes than those at 12.5 days in vivo. It is concluded that the digestive function and energy metabolism of the visceral yolk-sac epithelium are almost identical in vitro and in vivo at 10.5 and 11.5 days.

Keywords

Acid Phosphatase Isocitrate Succinate Dehydrogenase Mitochondrial Enzyme Histochemical Study 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Beck F (1981) Induced cell injury and cell death as a cause of congenital malformations in rats. Histochem J 13:667–679Google Scholar
  2. Beck F, Lowy A (1982) The effect of cathepsin inhibitor on rat embryos grown in vitro. J Embryol Exp Morphol 71:1–9Google Scholar
  3. Beck F, Lloyd JB, Griffiths A (1967a) Lysosomal enzyme inhibition by trypan blue: A theory of teratogenesis. Science 157:1080–1082Google Scholar
  4. Beck F, Lloyd JB, Griffiths A (1967b) A histochemical and biochemical study of some aspects of placental function in the rat using maternal injection of horseradish peroxidase. J Anat 101:461–478Google Scholar
  5. Berry CL (1968) Comparison of in vivo and in vitro growth of the rat foetus. Nature 219:92–93Google Scholar
  6. Brown NA, Fabro S (1981) Quantitation of rat embryonic development in vitro: A morphological scoring system. Teratology 24:65–78Google Scholar
  7. Cockroft DL (1973) Development in culture of rat foetuses explanted at 12.5 and 13.5 days of gestation. J Embryol Exp Morphol 29:473–483Google Scholar
  8. Cockroft DL (1976) Comparison of in vitro and in vivo development of rat foetuses. Dev Biol 48:163–172Google Scholar
  9. Freeman SJ, Lloyd JB (1983a) Evidences that protein ingested by the rat visceral yolk sac yielded amino acids for synthesis of embryonic protcin. J Embryol Exp Morphol 73:307–315Google Scholar
  10. Freeman SJ, Lloyd JB (1983b) Inhibition of proteolysis in rat yolk sac as a cause of teratogenesis. Effects of leupeptin in vitro and in vivo. J Embryol Exp Morphol 78:183–193Google Scholar
  11. Freeman SJ, Beck F, Lloyd JB (1981) The role of the visceral yolk sac in mediating protein utilization by rat embryos cultured in vitro. J Embryol Exp Morphol 66:223–234Google Scholar
  12. Freeman SJ, Brent RL, Lloyd JB (1982) The effect of teratogenic antiserum on yolk-sac function in rat embryos cultured in vitro. J Embryol Exp Morphol 71:63–74Google Scholar
  13. Gass P, Kugler P (1984) Enzymdifferenzierung des visceralen Dottersackepithels der Ratte. Z mikrosk-anat Forsch (in press)Google Scholar
  14. Goctze T, Franke H, Oswald B, Schlag B, Goetze E (1975) Effects of goat anti-rat placenta IgG on the in vitro uptake of 125I-labeled human serum albumin by the rat visceral yolk sac. Biol Neonate 27:221–231Google Scholar
  15. Gossrau R (1981) Investigation of proteases in the digestive tract using 4-methoxy-2-naphthylamine (MNA) substrates. J Histochem Cytochem 29:464–480Google Scholar
  16. Galamhusein AP, Moore WJ, Gupta M, Beck F (1982) Trypan blue teratogenesis in the rat: Further observations in vitro. Teratology 26:289–297Google Scholar
  17. Gupta M, Beck F (1983) Growth of 9.5-day rat embryos in human serum. J Embryol Exp Morphol 76:1–8Google Scholar
  18. Gupta M, Gulamhusein AP, Beck F (1982) Morphometric analysis of the visceral yolk sac endoderm in the rat in vivo and in vitro. J Reprod Fert 65:239–245Google Scholar
  19. Lear D, Clarke A, Gulamhusein AP, Huxham M, Beck F (1983) Morphological, total nucleic acid and total protein analyses of rat embryos cultured in supplemented and unsupplemented human serum. J Anat 137:279–285Google Scholar
  20. Lojda Z (1970) Indigogenic methods for glycosidases. II. An improved method for β-D-galactosidase and its application to localization studies of the enzymes in the intestine and in other tissues. Histochemie 23:266–288Google Scholar
  21. Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry. A laboratory manual. Springer, Berlin Heidelberg New YorkGoogle Scholar
  22. New DAT (1978) Whole-embryo culture and the study of mammalian embryos during organogenesis. Biol Rev 53:81–122Google Scholar
  23. New DAT, Brent RL (1972) Effect of yolk sac antibody on rat embryos grown in culture. J Embryol Exp Morphol 27:543–553Google Scholar
  24. New DAT, Coppola PT, Terry S (1973) Culture of explanted rat embryos in rotating tubes. J Reprod Fert 35:135–138Google Scholar
  25. New DAT, Coppola PT, Cockroft DL (1976a) Improved development of head-fold rat embryos in culture resulting from low oxygen and modifications of the culture serum. J Reprod Fert 48:219–222Google Scholar
  26. New DAT, Coppola PT, Cockroft DL (1976b) Comparison of growth in vitro and in vivo of post-implantation rat embryos. J Embryol Exp Morphol 36:133–144Google Scholar
  27. Novikoff AB, Goldfischer S (1969) Visualisation of peroxisomes (microbodies) and mitochondria with diaminobenzidine. J Histochem Cytochem 17:517–526Google Scholar
  28. Payne GS, Deuchar EM (1972) An in vitro study of functions of embryonic membranes in the rat. J Embryol Exp Morphol 27:533–542Google Scholar
  29. Rutenberg AM, Kim H, Fischbein JW, Hanker LS, Wasserkrug HL, Seligman AM (1969) γ-Glutamyl transpeptidase activity. J Histochem Cytochem 17:517–526Google Scholar
  30. Sharma R, Peel S (1979) Uptake of marker proteins by glycoprotein-containing cells of the pregnant rat uterus and placenta. J Anat 129:707–718Google Scholar
  31. Shepard TH, Tanimura T, Robkin MA (1970) Energy metabolism in early mammalian embryos. Dev Biol Suppl 4:42–58Google Scholar
  32. Steele CE, New DAT (1974) Serum variants causing the formation of double hearts and other abnormalities in explanted rat embryos. J Embryol Exp Morphol 31:707–719Google Scholar
  33. Williams KE, Roberts G, Kidston ME, Beck F, Lloyd JB (1976) Inhibition of pinocytosis in rat yolk sac by trypan blue. Teratology 14:343–354Google Scholar
  34. Winckler J (1970a) Zum Einfrieren von Geweben in Stickstoffgekühltem Propan. Histochemie 23:44–50Google Scholar
  35. Winckler J (1970b) Verwendung gefriergetrockneter Kryostatschnitte für histologische und histochemische Untersuchungen. Histochemie 24:168–186Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • A. Miki
    • 1
  • P. Kugler
    • 1
  1. 1.Institute of AnatomyUniversity of WürzburgWürzburgGermany

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