Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1988 Oct;82(4):1275–1281. doi: 10.1172/JCI113726

Gene for the rat atrial natriuretic peptide is regulated by glucocorticoids in vitro.

D G Gardner 1, B J Gertz 1, C F Deschepper 1, D Y Kim 1
PMCID: PMC442679  PMID: 2971674

Abstract

Glucocorticoids regulate the expression of the gene for atrial natriuretic peptide (ANP) in neonatal cardiocytes. Dexamethasone (Dex) increased cytoplasmic ANP mRNA levels and media ANP immunoreactivity in a dose-dependent fashion. These effects were not shared by the other classes of steroid hormones and were reversed by the glucocorticoid antagonist RU 38486. The effect on ANP mRNA levels resulted, at least in part, from enhanced transcription of the gene. Dex effected a two-fold increase in ANP gene activity assessed using a run-on transcription assay. The turnover of the ANP transcript was approximated using a standard pulse-chase technique. The half-life of the ANP mRNA was 18 h in hormone-free media. In the presence of Dex this half-life increased modestly to 30 h, although the increase relative to the control did not reach statistical significance. The effect of Dex at the level of the individual myocardial cell was assessed by in situ hybridization analysis using a specific [3H]cRNA probe. These studies demonstrated a significant level of ANP expression within a subpopulation of cells in the cultures. Exposure of the cells to Dex for 24 h did not recruit additional cells into the expressing pool (27.3% cells/high power field vs. 31.3% for the control) but did increase the level of expression (i.e., grain density) within individual cells. These findings indicate that glucocorticoids stimulate expression of the ANP gene directly at the level of the myocardial cell. This results predominantly from transcriptional activation in cells already expressing the gene rather than through recruitment of previously quiescent cells.

Full text

PDF
1275

Images in this article

1277Image
on p.1277
1277Image
on p.1277
1279Image
on p.1279
1279Image
on p.1279
1279Image
on p.1279
1279Image
on p.1279

Selected References

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

  1. Argentin S., Nemer M., Drouin J., Scott G. K., Kennedy B. P., Davies P. L. The gene for rat atrial natriuretic factor. J Biol Chem. 1985 Apr 25;260(8):4568–4571. [PubMed] [Google Scholar]
  2. Atarashi K., Mulrow P. J., Franco-Saenz R., Snajdar R., Rapp J. Inhibition of aldosterone production by an atrial extract. Science. 1984 Jun 1;224(4652):992–994. doi: 10.1126/science.6326267. [DOI] [PubMed] [Google Scholar]
  3. Bauer R. F., Arthur L. O., Fine D. L. Propagation of mouse mammary tumor cell lines and production of mouse mammary tumor virus in a serum-free medium. In Vitro. 1976 Aug;12(8):558–563. doi: 10.1007/BF02797439. [DOI] [PubMed] [Google Scholar]
  4. Bengele H. H., McNamara E. R., Alexander E. A. Natriuresis after adrenal enucleation: effect of spironolactone and dexamethasone. Am J Physiol. 1977 Jul;233(1):F8–12. doi: 10.1152/ajprenal.1977.233.1.F8. [DOI] [PubMed] [Google Scholar]
  5. Bloch K. D., Scott J. A., Zisfein J. B., Fallon J. T., Margolies M. N., Seidman C. E., Matsueda G. R., Homcy C. J., Graham R. M., Seidman J. G. Biosynthesis and secretion of proatrial natriuretic factor by cultured rat cardiocytes. Science. 1985 Dec 6;230(4730):1168–1171. doi: 10.1126/science.2933808. [DOI] [PubMed] [Google Scholar]
  6. Bloom E., Matulich D. T., Lan N. C., Higgins S. J., Simons S. S., Baxter J. D. Nuclear binding of glucocorticoid receptors: relations between cytosol binding, activation and the biological response. J Steroid Biochem. 1980 Jan;12:175–184. doi: 10.1016/0022-4731(80)90267-8. [DOI] [PubMed] [Google Scholar]
  7. Burnett J. C., Jr, Granger J. P., Opgenorth T. J. Effects of synthetic atrial natriuretic factor on renal function and renin release. Am J Physiol. 1984 Nov;247(5 Pt 2):F863–F866. doi: 10.1152/ajprenal.1984.247.5.F863. [DOI] [PubMed] [Google Scholar]
  8. Danesch U., Hashimoto S., Renkawitz R., Schütz G. Transcriptional regulation of the tryptophan oxygenase gene in rat liver by glucocorticoids. J Biol Chem. 1983 Apr 25;258(8):4750–4753. [PubMed] [Google Scholar]
  9. Day M. L., Schwartz D., Wiegand R. C., Stockman P. T., Brunnert S. R., Tolunay H. E., Currie M. G., Standaert D. G., Needleman P. Ventricular atriopeptin. Unmasking of messenger RNA and peptide synthesis by hypertrophy or dexamethasone. Hypertension. 1987 May;9(5):485–491. doi: 10.1161/01.hyp.9.5.485. [DOI] [PubMed] [Google Scholar]
  10. De Léan A., Racz K., Gutkowska J., Nguyen T. T., Cantin M., Genest J. Specific receptor-mediated inhibition by synthetic atrial natriuretic factor of hormone-stimulated steroidogenesis in cultured bovine adrenal cells. Endocrinology. 1984 Oct;115(4):1636–1638. doi: 10.1210/endo-115-4-1636. [DOI] [PubMed] [Google Scholar]
  11. Dobner P. R., Kawasaki E. S., Yu L. Y., Bancroft F. C. Thyroid or glucocorticoid hormone induces pre-growth-hormone mRNA and its probable nuclear precursor in rat pituitary cells. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2230–2234. doi: 10.1073/pnas.78.4.2230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garcia R., Debinski W., Gutkowska J., Kuchel O., Thibault G., Genest J., Cantin M. Gluco- and mineralocorticoids may regulate the natriuretic effect and the synthesis and release of atrial natriuretic factor by the rat atria in vivo. Biochem Biophys Res Commun. 1985 Sep 16;131(2):806–814. doi: 10.1016/0006-291x(85)91311-7. [DOI] [PubMed] [Google Scholar]
  13. Gardner D. G., Deschepper C. F., Ganong W. F., Hane S., Fiddes J., Baxter J. D., Lewicki J. Extra-atrial expression of the gene for atrial natriuretic factor. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6697–6701. doi: 10.1073/pnas.83.18.6697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gardner D. G., Hane S., Trachewsky D., Schenk D., Baxter J. D. Atrial natriuretic peptide mRNA is regulated by glucocorticoids in vivo. Biochem Biophys Res Commun. 1986 Sep 30;139(3):1047–1054. doi: 10.1016/s0006-291x(86)80283-2. [DOI] [PubMed] [Google Scholar]
  15. Grünfeld J. P., Eloy L., Moura A. M., Ganeval D., Ramos-Frendo B., Worcel M. Effects of antiglucocorticoids on glucocorticoid hypertension in the rat. Hypertension. 1985 Mar-Apr;7(2):292–299. doi: 10.1161/01.hyp.7.2.292. [DOI] [PubMed] [Google Scholar]
  16. Hager L. J., Palmiter R. D. Transcriptional regulation of mouse liver metallothionein-I gene by glucocorticoids. Nature. 1981 May 28;291(5813):340–342. doi: 10.1038/291340a0. [DOI] [PubMed] [Google Scholar]
  17. Kafatos F. C., Jones C. W., Efstratiadis A. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979 Nov 24;7(6):1541–1552. doi: 10.1093/nar/7.6.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
  19. Maki M., Takayanagi R., Misono K. S., Pandey K. N., Tibbetts C., Inagami T. Structure of rat atrial natriuretic factor precursor deduced from cDNA sequence. Nature. 1984 Jun 21;309(5970):722–724. doi: 10.1038/309722a0. [DOI] [PubMed] [Google Scholar]
  20. Matrisian L. M., Glaichenhaus N., Gesnel M. C., Breathnach R. Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts. EMBO J. 1985 Jun;4(6):1435–1440. doi: 10.1002/j.1460-2075.1985.tb03799.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matsubara H., Hirata Y., Yoshimi H., Takata S., Takagi Y., Iida T., Yamane Y., Umeda Y., Nishikawa M., Inada M. Effects of steroid and thyroid hormones on synthesis of atrial natriuretic peptide by cultured atrial myocytes of rat. Biochem Biophys Res Commun. 1987 May 29;145(1):336–343. doi: 10.1016/0006-291x(87)91326-x. [DOI] [PubMed] [Google Scholar]
  22. Matsubara H., Hirata Y., Yoshimi H., Takata S., Takagi Y., Yamane Y., Umeda Y., Nishikawa M., Inada M. Ventricular myocytes from neonatal rats are more responsive to dexamethasone than atrial myocytes in synthesis of atrial natriuretic peptide. Biochem Biophys Res Commun. 1987 Nov 13;148(3):1030–1038. doi: 10.1016/s0006-291x(87)80235-8. [DOI] [PubMed] [Google Scholar]
  23. Maxwell I. H., Van Ness J., Hahn W. E. Assay of DNA-RNA hybrids by S1 nuclease digestion and adsorption to DEAE-cellulose filters. Nucleic Acids Res. 1978 Jun;5(6):2033–2038. doi: 10.1093/nar/5.6.2033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  25. Nemer M., Chamberland M., Sirois D., Argentin S., Drouin J., Dixon R. A., Zivin R. A., Condra J. H. Gene structure of human cardiac hormone precursor, pronatriodilatin. Nature. 1984 Dec 13;312(5995):654–656. doi: 10.1038/312654a0. [DOI] [PubMed] [Google Scholar]
  26. Oikawa S., Imai M., Ueno A., Tanaka S., Noguchi T., Nakazato H., Kangawa K., Fukuda A., Matsuo H. Cloning and sequence analysis of cDNA encoding a precursor for human atrial natriuretic polypeptide. Nature. 1984 Jun 21;309(5970):724–726. doi: 10.1038/309724a0. [DOI] [PubMed] [Google Scholar]
  27. Payvar F., DeFranco D., Firestone G. L., Edgar B., Wrange O., Okret S., Gustafsson J. A., Yamamoto K. R. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell. 1983 Dec;35(2 Pt 1):381–392. doi: 10.1016/0092-8674(83)90171-x. [DOI] [PubMed] [Google Scholar]
  28. Samson W. K. Atrial natriuretic factor inhibits dehydration and hemorrhage-induced vasopressin release. Neuroendocrinology. 1985 Mar;40(3):277–279. doi: 10.1159/000124085. [DOI] [PubMed] [Google Scholar]
  29. Schwartz D., Geller D. M., Manning P. T., Siegel N. R., Fok K. F., Smith C. E., Needleman P. Ser-Leu-Arg-Arg-atriopeptin III: the major circulating form of atrial peptide. Science. 1985 Jul 26;229(4711):397–400. doi: 10.1126/science.3160114. [DOI] [PubMed] [Google Scholar]
  30. Seidman C. E., Bloch K. D., Klein K. A., Smith J. A., Seidman J. G. Nucleotide sequences of the human and mouse atrial natriuretic factor genes. Science. 1984 Dec 7;226(4679):1206–1209. doi: 10.1126/science.6542248. [DOI] [PubMed] [Google Scholar]
  31. Seidman C. E., Duby A. D., Choi E., Graham R. M., Haber E., Homcy C., Smith J. A., Seidman J. G. The structure of rat preproatrial natriuretic factor as defined by a complementary DNA clone. Science. 1984 Jul 20;225(4659):324–326. doi: 10.1126/science.6234658. [DOI] [PubMed] [Google Scholar]
  32. Simpson P., Savion S. Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells. Cross-striations, ultrastructure, and chronotropic response to isoproterenol. Circ Res. 1982 Jan;50(1):101–116. doi: 10.1161/01.res.50.1.101. [DOI] [PubMed] [Google Scholar]
  33. Slater E. P., Rabenau O., Karin M., Baxter J. D., Beato M. Glucocorticoid receptor binding and activation of a heterologous promoter by dexamethasone by the first intron of the human growth hormone gene. Mol Cell Biol. 1985 Nov;5(11):2984–2992. doi: 10.1128/mcb.5.11.2984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Spindler S. R., Mellon S. H., Baxter J. D. Growth hormone gene transcription is regulated by thyroid and glucocorticoid hormones in cultured rat pituitary tumor cells. J Biol Chem. 1982 Oct 10;257(19):11627–11632. [PubMed] [Google Scholar]
  35. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yamanaka M., Greenberg B., Johnson L., Seilhamer J., Brewer M., Friedemann T., Miller J., Atlas S., Laragh J., Lewicki J. Cloning and sequence analysis of the cDNA for the rat atrial natriuretic factor precursor. Nature. 1984 Jun 21;309(5970):719–722. doi: 10.1038/309719a0. [DOI] [PubMed] [Google Scholar]
  37. de Bold A. J., Borenstein H. B., Veress A. T., Sonnenberg H. A rapid and potent natriuretic response to intravenous injection of atrial myocardial extract in rats. Life Sci. 1981 Jan 5;28(1):89–94. doi: 10.1016/0024-3205(81)90370-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES