Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Jun;77(6):3524–3528. doi: 10.1073/pnas.77.6.3524

Cultured mouse embryos metabolize benzo[a]pyrene during early gestation: genetic differences detectable by sister chromatid exchange.

S M Galloway, P E Perry, J Meneses, D W Nebert, R A Pedersen
PMCID: PMC349649  PMID: 6932035

Abstract

Mouse embryos explanted at 7 1/2 or 8 1/2 days of gestation were cultured in medium containing benzo[a]pyrene and supplemented with 5-bromodeoxyuridine to allow detection of sister chromatid exchanges. The murine Ah locus regulates the inducible metabolism of polycyclic hydrocarbons such as benzo[a]pyrene. A high frequency of sister chromatid exchange was induced by benzo[a]pyrene in embryos from three Ah-"responsive" inbred strains (BALB/cDub, C3H/AnfCum, and C57BL/6N); there was little or no increase in two Ah-"nonresponsive" inbred strains (AKR/J and DBA/2J). Benzo[a]pyrene also induced sister chromatid exchanges in the Ah-responsive recombinant inbred line B6NXAKN-12 but not in the Ah-nonresponsive recombinant inbred line B6NXAKN-3. Sister chromatid exchange in cultured Ah-responsive mouse embryos was thus shown to be a sensitive assay. These data provide direct evidence that genetically responsive mouse embryos (early postimplantation stage) possess the subcellular processes necessary for induction of enzymes that metabolize benzo[a]pyrene to its chemically active forms(s). Both the Ah regulatory gene product (a cytoslic receptor) and the structural gene product (inducible cytochrome P1-450) therefore appear to be functional at an early embryonic age. Furthermore, this metabolic capacity may play an important role in the damage to embryonic cells by polycyclic hydracarbons.

Full text

PDF
3524

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ames B. N., Mccann J., Yamasaki E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat Res. 1975 Dec;31(6):347–364. doi: 10.1016/0165-1161(75)90046-1. [DOI] [PubMed] [Google Scholar]
  2. Atlas S. A., Taylor B. A., Diwan B. A., Nebert D. W. Inducible monooxygenase activities and 3-methyl-cholanthrene-initiated tumorigenesis in mouse recombinant inbred sublines. Genetics. 1976 Jul;83(3 PT2):537–550. [PMC free article] [PubMed] [Google Scholar]
  3. Bailey D. W. Recombinant-inbred strains. An aid to finding identity, linkage, and function of histocompatibility and other genes. Transplantation. 1971 Mar;11(3):325–327. doi: 10.1097/00007890-197103000-00013. [DOI] [PubMed] [Google Scholar]
  4. Boobis A. R., Nebert D. W., Felton J. S. Comparison of beta-naphthoflavone and 3-methylcholanthrene as inducers of hepatic cytochrome(s) P-448 and aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity. Mol Pharmacol. 1977 Mar;13(2):259–268. [PubMed] [Google Scholar]
  5. Chouroulinkov I., Gentil A., Grover P. L., Sims P. Tumour-initiating activities on mouse skin of dihydrodiols derived from benzo[a]pyrene. Br J Cancer. 1976 Nov;34(5):523–532. doi: 10.1038/bjc.1976.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Currie A. R., Bird C. C., Crawford A. M., Sims P. Embryopathic effects of 7,12-dimethylbenz(a)anthracene and its hydroxymethyl derivatives in the Sprague-Dawley rat. Nature. 1970 Jun 6;226(5249):911–914. doi: 10.1038/226911a0. [DOI] [PubMed] [Google Scholar]
  7. Evans E. P., Burtenshaw M. D., Ford C. E. Chromosomes of mouse embryos and newborn young: preparations from membranes and tail tips. Stain Technol. 1972 Sep;47(5):229–234. doi: 10.3109/10520297209116541. [DOI] [PubMed] [Google Scholar]
  8. Goto K., Akematsu T., Shimazu H., Sugiyama T. Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining. Chromosoma. 1975 Dec 10;53(3):223–230. doi: 10.1007/BF00329173. [DOI] [PubMed] [Google Scholar]
  9. Goto K., Maeda S., Kano Y., Sugiyama T. Factors involved in differential Giemsa-staining of sister chromatids. Chromosoma. 1978 May 16;66(4):351–359. doi: 10.1007/BF00328535. [DOI] [PubMed] [Google Scholar]
  10. Ikushima T., Wolff S. Sister chromatid exchanges induced by light flashes to 5-bromodeoxyuridine- and 5-iododeoxyuridine substituted Chinese hamster chromosomes. Exp Cell Res. 1974 Jul;87(1):15–19. doi: 10.1016/0014-4827(74)90521-7. [DOI] [PubMed] [Google Scholar]
  11. Kahl G. F., Friederici D. E., Bigelow S. W., Okey A. B., Nebert D. W. Ontogenetic expression of regulatory and structural gene products associated with the Ah locus. Comparison of rat, mouse, rabbit and Sigmoden hispedis. Dev Pharmacol Ther. 1980;1(2-3):137–162. [PubMed] [Google Scholar]
  12. Kram D., Schneider E. L. Reduced frequencies of mitomycin-C induced sister chromatid exchanges in AKR mice. Hum Genet. 1978 Feb 23;41(1):45–51. doi: 10.1007/BF00278870. [DOI] [PubMed] [Google Scholar]
  13. Lambert G. H., Nebert D. W. Genetically mediated induction of drug-metabolizing enzymes associated with congenital defects in the mouse. Teratology. 1977 Oct;16(2):147–153. doi: 10.1002/tera.1420160206. [DOI] [PubMed] [Google Scholar]
  14. Latt S. A. Sister chromatid exchanges, indices of human chromosome damage and repair: detection by fluorescence and induction by mitomycin C. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3162–3166. doi: 10.1073/pnas.71.8.3162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Malaveille C., Bartsch H., Grover P. L., Sims P. Mutagenicity of non-K-region diols and diol-epoxides of benz(a)anthracene and benzo(a)pyrene in S. typhimurium TA 100. Biochem Biophys Res Commun. 1975 Sep 16;66(2):693–700. doi: 10.1016/0006-291x(75)90565-3. [DOI] [PubMed] [Google Scholar]
  16. Marquardt H., Grover P. L., Sims P. In vitro malignant transformation of mouse fibroblasts by non-K-region dihydrodiols derived from 7-methylbenz(a)anthracene, 7,12-dimethylbenz(a)anthracene, and benzo(a)pyrene. Cancer Res. 1976 Jun;36(6):2059–2064. [PubMed] [Google Scholar]
  17. Nebert D. W., Bausserman L. L. Genetic differences in the extent of aryl hydrocarbon hydroxylase induction in mouse fetal cell cultures. J Biol Chem. 1970 Dec 10;245(23):6373–6382. [PubMed] [Google Scholar]
  18. Nebert D. W., Gelboin H. V. Substrate-inducible microsomal aryl hydroxylase in mammalian cell culture. II. Cellular responses during enzyme induction. J Biol Chem. 1968 Dec 10;243(23):6250–6261. [PubMed] [Google Scholar]
  19. Nebert D. W., Gelboin H. V. The in vivo and in vitro induction of aryl hydrocarbon hydroxylase in mammalian cells of different species, tissues, strains, and developmental and hormonal states. Arch Biochem Biophys. 1969 Oct;134(1):76–89. doi: 10.1016/0003-9861(69)90253-7. [DOI] [PubMed] [Google Scholar]
  20. Nebert D. W., Goujon F. M., Gielen J. E. Aryl hydrocarbon hydroxylase induction by polycyclic hydrocarbons: simple autosomal dominant trait in the mouse. Nat New Biol. 1972 Mar 29;236(65):107–110. doi: 10.1038/newbio236107a0. [DOI] [PubMed] [Google Scholar]
  21. Nebert D. W., Jensen N. M. The Ah locus: genetic regulation of the metabolism of carcinogens, drugs, and other environmental chemicals by cytochrome P-450-mediated monooxygenases. CRC Crit Rev Biochem. 1979;6(4):401–437. doi: 10.3109/10409237909105427. [DOI] [PubMed] [Google Scholar]
  22. Okey A. B., Bondy G. P., Mason M. E., Kahl G. F., Eisen H. J., Guenthner T. M., Nebert D. W. Regulatory gene product of the Ah locus. Characterization of the cytosolic inducer-receptor complex and evidence for its nuclear translocation. J Biol Chem. 1979 Nov 25;254(22):11636–11648. [PubMed] [Google Scholar]
  23. Pal K., Tierney B., Grover P. L., Sims P. Induction of sister-chromatid exchanges in Chinese hamster ovary cells treated in vitro with non-K-region dihydrodiols of 7-methylbenz[a]anthracene and benzo[a]pyrene. Mutat Res. 1978 Jun;50(3):367–375. doi: 10.1016/0027-5107(78)90041-6. [DOI] [PubMed] [Google Scholar]
  24. Perry P., Wolff S. New Giemsa method for the differential staining of sister chromatids. Nature. 1974 Sep 13;251(5471):156–158. doi: 10.1038/251156a0. [DOI] [PubMed] [Google Scholar]
  25. Popescu N. C., Turnbull D., DiPaolo J. A. Sister chromatid exchange and chromosome aberration analysis with the use of several carcinogens and noncarcinogens. J Natl Cancer Inst. 1977 Jul;59(1):289–293. doi: 10.1093/jnci/59.1.289. [DOI] [PubMed] [Google Scholar]
  26. Rigdon R. H., Rennels E. G. Effect of feeding benzpyrene on reproduction in the rat. Experientia. 1964 Apr 15;20(4):224–226. doi: 10.1007/BF02135417. [DOI] [PubMed] [Google Scholar]
  27. Rudiger H. W., Kohl F., Mangels W., von Wichert P., Bartram C. R., Wohler W., Passarge E. Benzpyrene induces sister chromatid exchanges in cultured human lymphocytes. Nature. 1976 Jul 22;262(5566):290–292. doi: 10.1038/262290a0. [DOI] [PubMed] [Google Scholar]
  28. SAVKUR L. D., BATRA B. K., SRIDHARAN B. N., SANGHVI L. D. Effect of 20-methylcholanthrene on mouse embryos. III. Strain C3H(Jax)--by caesarean section. J Reprod Fertil. 1962 Jun;3:422–424. doi: 10.1530/jrf.0.0030422. [DOI] [PubMed] [Google Scholar]
  29. Schönwald A. D., Bartram C. R., Rüdiger H. W. Benzpyrene-induced sister chromatid exchanges in lymphocytes of patients with lung cancer. Hum Genet. 1977 May 10;36(3):261–264. doi: 10.1007/BF00446274. [DOI] [PubMed] [Google Scholar]
  30. Shum S., Jensen N. M., Nebert D. W. The murine Ah locus: in utero toxicity and teratogenesis associated with genetic differences in benzo[a]pyrene metabolism. Teratology. 1979 Dec;20(3):365–376. doi: 10.1002/tera.1420200307. [DOI] [PubMed] [Google Scholar]
  31. Sims P., Grover P. L., Swaisland A., Pal K., Hewer A. Metabolic activation of benzo(a)pyrene proceeds by a diol-epoxide. Nature. 1974 Nov 22;252(5481):326–328. doi: 10.1038/252326a0. [DOI] [PubMed] [Google Scholar]
  32. Swank R. T., Bailey D. W. Recombinant inbred lines: value in the genetic analysis of biochemical variants. Science. 1973 Sep 28;181(4106):1249–1252. doi: 10.1126/science.181.4106.1249. [DOI] [PubMed] [Google Scholar]
  33. Takehisa S., Wolff S. Induction of sister chromatid exchanges in Chinese hamster cells by carcinogenic mutagens requiring metabolic activation. Mutat Res. 1977 Nov;45(2):263–270. doi: 10.1016/0027-5107(77)90026-4. [DOI] [PubMed] [Google Scholar]
  34. Tomatis L., Turusov V., Guibbert D., Duperray B., Malaveille C., Pacheco H. Transplacental carcinogenic effect of 3-methylcholanthrene in mice and its quantitation in fetal tissues. J Natl Cancer Inst. 1971 Sep;47(3):645–651. [PubMed] [Google Scholar]
  35. Wiley L. M., Pedersen R. A. Morphology of mouse egg cylinder development in vitro: a light and electron microscopic study. J Exp Zool. 1977 Jun;200(3):389–402. doi: 10.1002/jez.1402000309. [DOI] [PubMed] [Google Scholar]
  36. Wolff S. Sister chromatid exchange. Annu Rev Genet. 1977;11:183–201. doi: 10.1146/annurev.ge.11.120177.001151. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES