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
. 1993 Aug 1;90(15):7322–7326. doi: 10.1073/pnas.90.15.7322

Thyroid hormone-dependent transcriptional regulation of exogenous genes transferred into Xenopus tadpole muscle in vivo.

A de Luze 1, L Sachs 1, B Demeneix 1
PMCID: PMC47129  PMID: 8346251

Abstract

Metamorphosis in amphibians is marked by dramatic thyroid hormone-induced changes that include tail regression. To examine thyroid hormone effects on gene transcription during the early stages of tail resorption, we injected exogenous genes directly into the caudal skeletal muscle of Xenopus tadpoles and followed their expression in vivo. Gene expression was both strong and reproducible, and it correlated with the amount of DNA injected. Moreover, expression continued as long as the animals were blocked in prometamorphosis by antithyroid drugs (for up to 4 months). Thyroid hormone-dependent effects on transcription were examined by using a palindromic thyroid hormone response element linked to a chloramphenicol acetyltransferase reporter gene. Reporter gene expressions were normalized for transfection efficiency by using a constitutively expressed luciferase construct. Physiological concentrations of 3,5,3' triiodo-L-thyronine (1 nM), applied for 120 hr, produced a 5-fold increase in transcription (P < 0.05) from the thyroid hormone response element but did not modify transcription from constitutive viral promoters. This study thus demonstrates that by directly expressing genes in Xenopus tadpole muscle in vivo, one can exploit the powerful experimental advantages of gene transfer systems in an intact, physiologically normal animal.

Full text

PDF
7322

Images in this article

7324Fig. 1
on p.7324
7324Fig. 2
on p.7324

Selected References

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

  1. Acsadi G., Dickson G., Love D. R., Jani A., Walsh F. S., Gurusinghe A., Wolff J. A., Davies K. E. Human dystrophin expression in mdx mice after intramuscular injection of DNA constructs. Nature. 1991 Aug 29;352(6338):815–818. doi: 10.1038/352815a0. [DOI] [PubMed] [Google Scholar]
  2. Acsadi G., Jiao S. S., Jani A., Duke D., Williams P., Chong W., Wolff J. A. Direct gene transfer and expression into rat heart in vivo. New Biol. 1991 Jan;3(1):71–81. [PubMed] [Google Scholar]
  3. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  4. Baker B. S., Tata J. R. Accumulation of proto-oncogene c-erb-A related transcripts during Xenopus development: association with early acquisition of response to thyroid hormone and estrogen. EMBO J. 1990 Mar;9(3):879–885. doi: 10.1002/j.1460-2075.1990.tb08185.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baker B. S., Tata J. R. Prolactin prevents the autoinduction of thyroid hormone receptor mRNAs during amphibian metamorphosis. Dev Biol. 1992 Feb;149(2):463–467. doi: 10.1016/0012-1606(92)90301-v. [DOI] [PubMed] [Google Scholar]
  6. Bugge T. H., Pohl J., Lonnoy O., Stunnenberg H. G. RXR alpha, a promiscuous partner of retinoic acid and thyroid hormone receptors. EMBO J. 1992 Apr;11(4):1409–1418. doi: 10.1002/j.1460-2075.1992.tb05186.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Glass C. K., Franco R., Weinberger C., Albert V. R., Evans R. M., Rosenfeld M. G. A c-erb-A binding site in rat growth hormone gene mediates trans-activation by thyroid hormone. Nature. 1987 Oct 22;329(6141):738–741. doi: 10.1038/329738a0. [DOI] [PubMed] [Google Scholar]
  8. Glass C. K., Holloway J. M. Regulation of gene expression by the thyroid hormone receptor. Biochim Biophys Acta. 1990 Dec 11;1032(2-3):157–176. doi: 10.1016/0304-419x(90)90002-i. [DOI] [PubMed] [Google Scholar]
  9. Kawahara A., Baker B. S., Tata J. R. Developmental and regional expression of thyroid hormone receptor genes during Xenopus metamorphosis. Development. 1991 Aug;112(4):933–943. doi: 10.1242/dev.112.4.933. [DOI] [PubMed] [Google Scholar]
  10. Lezoualc'h F., Hassan A. H., Giraud P., Loeffler J. P., Lee S. L., Demeneix B. A. Assignment of the beta-thyroid hormone receptor to 3,5,3'-triiodothyronine-dependent inhibition of transcription from the thyrotropin-releasing hormone promoter in chick hypothalamic neurons. Mol Endocrinol. 1992 Nov;6(11):1797–1804. doi: 10.1210/mend.6.11.1480171. [DOI] [PubMed] [Google Scholar]
  11. Machuca I., Tata J. R. Autoinduction of thyroid hormone receptor during metamorphosis is reproduced in Xenopus XTC-2 cells. Mol Cell Endocrinol. 1992 Sep;87(1-3):105–113. doi: 10.1016/0303-7207(92)90238-2. [DOI] [PubMed] [Google Scholar]
  12. Oppenheimer J. H., Schwartz H. L., Surks M. I., Koerner D., Dillmann W. H. Nuclear receptors and the initiation of thyroid hormone action. Recent Prog Horm Res. 1976;32:529–565. doi: 10.1016/b978-0-12-571132-6.50029-4. [DOI] [PubMed] [Google Scholar]
  13. Ribeiro R. C., Kushner P. J., Apriletti J. W., West B. L., Baxter J. D. Thyroid hormone alters in vitro DNA binding of monomers and dimers of thyroid hormone receptors. Mol Endocrinol. 1992 Jul;6(7):1142–1152. doi: 10.1210/mend.6.7.1508227. [DOI] [PubMed] [Google Scholar]
  14. Rubio R., Sperelakis N. Penetration of horseradish peroxidase into the terminal cisternae of frog skeletal muscle fibers and blockade of caffeine contracture by Ca ++ depletion. Z Zellforsch Mikrosk Anat. 1972;124(1):57–71. [PubMed] [Google Scholar]
  15. Saleem M., Atkinson B. G. Thyroid hormone-induced regulation of protein synthesis in tadpole tail muscle. Gen Comp Endocrinol. 1979 Aug;38(4):441–450. doi: 10.1016/0016-6480(79)90151-5. [DOI] [PubMed] [Google Scholar]
  16. Sap J., Muñoz A., Damm K., Goldberg Y., Ghysdael J., Leutz A., Beug H., Vennström B. The c-erb-A protein is a high-affinity receptor for thyroid hormone. Nature. 1986 Dec 18;324(6098):635–640. doi: 10.1038/324635a0. [DOI] [PubMed] [Google Scholar]
  17. Umesono K., Murakami K. K., Thompson C. C., Evans R. M. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991 Jun 28;65(7):1255–1266. doi: 10.1016/0092-8674(91)90020-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wahlström G. M., Sjöberg M., Andersson M., Nordström K., Vennström B. Binding characteristics of the thyroid hormone receptor homo- and heterodimers to consensus AGGTCA repeat motifs. Mol Endocrinol. 1992 Jul;6(7):1013–1022. doi: 10.1210/mend.6.7.1324417. [DOI] [PubMed] [Google Scholar]
  19. Weinberger C., Thompson C. C., Ong E. S., Lebo R., Gruol D. J., Evans R. M. The c-erb-A gene encodes a thyroid hormone receptor. Nature. 1986 Dec 18;324(6098):641–646. doi: 10.1038/324641a0. [DOI] [PubMed] [Google Scholar]
  20. Wolff J. A., Malone R. W., Williams P., Chong W., Acsadi G., Jani A., Felgner P. L. Direct gene transfer into mouse muscle in vivo. Science. 1990 Mar 23;247(4949 Pt 1):1465–1468. doi: 10.1126/science.1690918. [DOI] [PubMed] [Google Scholar]
  21. Yaoita Y., Shi Y. B., Brown D. D. Xenopus laevis alpha and beta thyroid hormone receptors. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7090–7094. doi: 10.1073/pnas.87.18.7090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yen P. M., Darling D. S., Carter R. L., Forgione M., Umeda P. K., Chin W. W. Triiodothyronine (T3) decreases binding to DNA by T3-receptor homodimers but not receptor-auxiliary protein heterodimers. J Biol Chem. 1992 Feb 25;267(6):3565–3568. [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