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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
. 1987 Oct;84(19):6760–6764. doi: 10.1073/pnas.84.19.6760

Localization of atrial natriuretic peptide mRNA and immunoreactivity in the rat heart and human atrial appendage.

Q Hamid 1, J Wharton 1, G Terenghi 1, C J Hassall 1, J Aimi 1, K M Taylor 1, H Nakazato 1, J E Dixon 1, G Burnstock 1, J M Polak 1
PMCID: PMC299164  PMID: 2958847

Abstract

The localization of mRNA encoding preproatrial natriuretic peptide was investigated in tissue sections and cultures of rat heart and in sections of human right atrial appendage using the technique of in situ hybridization with 32P- and 35S-labeled RNA probes. Rat atrial natriuretic peptide (ANP) transcripts were demonstrated in numerous atrial myocytes and, to a lesser extent, in ventricular myocytes in both tissue sections and newborn rat heart cultures. These findings are consistent with those obtained by RNA blot analysis of rat heart total RNA, indicating that a single prepro-ANP transcript of approximately 900 nucleotides was present in the ventricles as well as the atria. Using a 35S-labeled RNA probe for human ANP mRNA, ANP transcripts were also localized to the majority of myocytes in the human right atrial appendage. Only background levels of autoradiographic labeling were obtained when RNA probes identical to the coding sequence of rat or human ANP mRNA were used. A close correlation was found between the distribution of ANP immunoreactivity and prepro-ANP mRNA in these preparations. These results provide unequivocal evidence for the expression of the ANP gene in the rat ventricles, as well as the atria, because myocytes in these tissues have been established as the sites of both ANP localization and precursor biosynthesis. The combined use of cardiac cultures and in situ hybridization may be of value in future studies investigating the regulation of ANP synthesis in cardiac myocytes.

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Selected References

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  1. Anderson J. V., Christofides N. D., Vinas P., Wharton J., Varndell I. M., Polak J. M., Bloom S. R. Radioimmunoassay of alpha rat atrial natriuretic peptide. Neuropeptides. 1986 Feb-Mar;7(2):159–173. doi: 10.1016/0143-4179(86)90092-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Back H., Stumpf W. E., Ando E., Nokihara K., Forssmann W. G. Immunocytochemical evidence for CDD/ANP-like peptides in strands of myoendocrine cells associated with the ventricular conduction system of the rat heart. Anat Embryol (Berl) 1986;175(2):223–226. doi: 10.1007/BF00389598. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. COONS A. H., LEDUC E. H., CONNOLLY J. M. Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J Exp Med. 1955 Jul 1;102(1):49–60. doi: 10.1084/jem.102.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cantin M., Dagenais N., Salmi L., Gutkowska J., Ballak M., Thibault G., Garcia R., Genest J. Secretory patterns of atrial natriuretic factor (ANF) by cultured cardiocytes of right and left atrium from newborn and adult rats. Clin Exp Hypertens A. 1985;7(5-6):685–705. doi: 10.3109/10641968509077221. [DOI] [PubMed] [Google Scholar]
  7. Cantin M., Gutkowska J., Thibault G., Milne R. W., Ledoux S., MinLi S., Chapeau C., Garcia R., Hamet P., Genest J. Immunocytochemical localization of atrial natriuretic factor in the heart and salivary glands. Histochemistry. 1984;80(2):113–127. doi: 10.1007/BF00679984. [DOI] [PubMed] [Google Scholar]
  8. Chapeau C., Gutkowska J., Schiller P. W., Milne R. W., Thibault G., Garcia R., Genest J., Cantin M. Localization of immunoreactive synthetic atrial natriuretic factor (ANF) in the heart of various animal species. J Histochem Cytochem. 1985 Jun;33(6):541–550. doi: 10.1177/33.6.3158698. [DOI] [PubMed] [Google Scholar]
  9. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  10. Coghlan J. P., Penschow J. D., Hudson P. J., Niall H. D. Hybridization histochemistry: use of recombinant DNA for tissue localizations of specific mRNA populations. Clin Exp Hypertens A. 1984;6(1-2):63–78. doi: 10.3109/10641968409062551. [DOI] [PubMed] [Google Scholar]
  11. Forssmann W. G. Cardiac hormones. I. Review on the morphology, biochemistry, and molecular biology of the endocrine heart. Eur J Clin Invest. 1986 Dec;16(6):439–451. doi: 10.1111/j.1365-2362.1986.tb02159.x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Gee C. E., Roberts J. L. In situ hybridization histochemistry: a technique for the study of gene expression in single cells. DNA. 1983;2(2):157–163. doi: 10.1089/dna.1983.2.157. [DOI] [PubMed] [Google Scholar]
  14. Greenberg B. D., Bencen G. H., Seilhamer J. J., Lewicki J. A., Fiddes J. C. Nucleotide sequence of the gene encoding human atrial natriuretic factor precursor. Nature. 1984 Dec 13;312(5995):656–658. doi: 10.1038/312656a0. [DOI] [PubMed] [Google Scholar]
  15. Hassall C. J., Burnstock G. Intrinsic neurones and associated cells of the guinea-pig heart in culture. Brain Res. 1986 Jan 29;364(1):102–113. doi: 10.1016/0006-8993(86)90991-1. [DOI] [PubMed] [Google Scholar]
  16. Hoefler H., Childers H., Montminy M. R., Lechan R. M., Goodman R. H., Wolfe H. J. In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes. Histochem J. 1986 Nov-Dec;18(11-12):597–604. doi: 10.1007/BF01675295. [DOI] [PubMed] [Google Scholar]
  17. Huang W. M., Gibson S. J., Facer P., Gu J., Polak J. M. Improved section adhesion for immunocytochemistry using high molecular weight polymers of L-lysine as a slide coating. Histochemistry. 1983;77(2):275–279. doi: 10.1007/BF00506570. [DOI] [PubMed] [Google Scholar]
  18. Kangawa K., Tawaragi Y., Oikawa S., Mizuno A., Sakuragawa Y., Nakazato H., Fukuda A., Minamino N., Matsuo H. Identification of rat gamma atrial natriuretic polypeptide and characterization of the cDNA encoding its precursor. Nature. 1984 Nov 8;312(5990):152–155. doi: 10.1038/312152a0. [DOI] [PubMed] [Google Scholar]
  19. Lattion A. L., Michel J. B., Arnauld E., Corvol P., Soubrier F. Myocardial recruitment during ANF mRNA increase with volume overload in the rat. Am J Physiol. 1986 Nov;251(5 Pt 2):H890–H896. doi: 10.1152/ajpheart.1986.251.5.H890. [DOI] [PubMed] [Google Scholar]
  20. Maki M., Parmentier M., Inagami T. Cloning of genomic DNA for human atrial natriuretic factor. Biochem Biophys Res Commun. 1984 Dec 14;125(2):797–802. doi: 10.1016/0006-291x(84)90609-0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Minth C. D., Bloom S. R., Polak J. M., Dixon J. E. Cloning, characterization, and DNA sequence of a human cDNA encoding neuropeptide tyrosine. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4577–4581. doi: 10.1073/pnas.81.14.4577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nakayama K., Ohkubo H., Hirose T., Inayama S., Nakanishi S. mRNA sequence for human cardiodilatin-atrial natriuretic factor precursor and regulation of precursor mRNA in rat atria. Nature. 1984 Aug 23;310(5979):699–701. doi: 10.1038/310699a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. ROSE G. A separable and multipurpose tissue culture chamber. Tex Rep Biol Med. 1954;12(4):1074–1083. [PubMed] [Google Scholar]
  27. 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]
  28. Siegel R. E., Young W. S., 3rd Detection of preprocholecystokinin and preproenkephalin A mRNAs in rat brain by hybridization histochemistry using complementary RNA probes. Neuropeptides. 1985 Dec;6(6):573–580. doi: 10.1016/0143-4179(85)90121-0. [DOI] [PubMed] [Google Scholar]
  29. Sylvestre D. L., Zisfein J. B., Graham R. M., Homcy C. J. Serum-mediated enhancement of ANF accumulation in the culture medium of cardiac atriocytes. Biochem Biophys Res Commun. 1986 Oct 15;140(1):151–159. doi: 10.1016/0006-291x(86)91070-3. [DOI] [PubMed] [Google Scholar]
  30. Takayanagi R., Tanaka I., Maki M., Inagami T. Effects of changes in water-sodium balance on levels of atrial natriuretic factor messenger RNA and peptide in rats. Life Sci. 1985 May 13;36(19):1843–1848. doi: 10.1016/0024-3205(85)90157-2. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Thompson R. P., Simson J. A., Currie M. G. Atriopeptin distribution in the developing rat heart. Anat Embryol (Berl) 1986;175(2):227–233. doi: 10.1007/BF00389599. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Yang-Feng T. L., Floyd-Smith G., Nemer M., Drouin J., Francke U. The pronatriodilatin gene is located on the distal short arm of human chromosome 1 and on mouse chromosome 4. Am J Hum Genet. 1985 Nov;37(6):1117–1128. [PMC free article] [PubMed] [Google Scholar]
  35. Zivin R. A., Condra J. H., Dixon R. A., Seidah N. G., Chrétien M., Nemer M., Chamberland M., Drouin J. Molecular cloning and characterization of DNA sequences encoding rat and human atrial natriuretic factors. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6325–6329. doi: 10.1073/pnas.81.20.6325. [DOI] [PMC free article] [PubMed] [Google Scholar]

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