Gene expression and atrial natriuretic factor processing and secretion in cultured at-1 cardiac myocytes

Nicholas A. Lanson, Christopher C. Glembotski, Mark E. Steinhelper, Loren J. Field, William C. Claycomb

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

Background. Studies were carried out to characterize several biochemical features of cultured AT-1 cells. Methods and Results. These cells were obtained from a transplantable atrial cardiomyocyte tumor lineage. Reverse transcriptase-polymerase chain reaction-based analyses demonstrated that the pattern of gene expression of cultured AT-1 cells was similar to that of adult atrial myocytes. AT-1 cells expressed atrial natriuretic factor (ANF), α-cardiac myosin heavy chain, α-cardiac actin, and connexin43. Radioimmunoassays verified that the cells synthesized, stored, and secreted ANF. Through size-exclusion, reversed-phase, and carboxymethyl-ion-exchange high-performance liquid chromatography, it was shown that cultured AT-1 cells stored ANF as pro-ANF (ANF-[1-126]), which was cosecretionally processed quantitatively to ANF-(1-98) and the bioactive 28-amino-acid ANF-(99-126). In addition, cultured AT-1 cells secreted ANF at almost a sixfold greater rate in response to endothelin-1, a potent secretagogue of ANF. KCl, metenkephalinamide, isoproterenol, phenylephrine, and 12-O-tetradecanoyl-phorbol-13-acetate also stimulated ANF release. Conclusions. These studies, in combination with previous findings, demonstrated that cultured AT-1 cells, while maintaining the ability to proliferate, have retained functional, biochemical, and ultrastructural features that are characteristic of adult atrial myocytes.

Original languageEnglish (US)
Pages (from-to)1835-1841
Number of pages7
JournalCirculation
Volume85
Issue number5
DOIs
StatePublished - May 1992
Externally publishedYes

Keywords

  • Cardiac myocyte hyperplasia
  • Cell culture
  • SV40 T antigen
  • Transgenic mice

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

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