Role of transiently altered sarcolemmal membrane permeability and basic fibroblast growth factor release in the hypertrophic response of adult rat ventricular myocytes to increased mechanical activity in vitro

David Kaye, David Pimental, Sanjay Prasad, Tiina Mäki, Hans Jörg Berger, Paul L. McNeil, Thomas W. Smith, Ralph A. Kelly

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

One of the trophic factors that has been implicated in initiating or facilitating growth in response to increased mechanical stress in several tissues and cell types is basic fibroblast growth factor (bFGF; FGF-2). Although mammalian cardiac muscle cells express bFGF, it is not known whether it plays a role in mediating cardiac adaptation to increased load, nor how release of the cytosolic 18-kD isoform of bFGF would be regulated in response to increased mechanical stress. To test the hypothesis that increased mechanical activity induces transient alterations in sarcolemmal permeability that allow cytosolic bFGF to be released and subsequently to act as an autocrine and paracrine growth stimulus, we examined primary isolates of adult rat ventricular myocytes maintained in serum-free, defined medium that were continually paced at 3 Hz for up to 5 d. Paced myocytes, but not nonpaced control cells, exhibited a "hypertrophic" response, which was characterized by increases in the rate of phenylalanine incorporation, total cellular pro-tein content, and cell size. These changes could be mimicked in control cells by exogenous recombinant bFGF and could be blocked in continually paced cells by a specific neutralizing anti-bFGF antibody. In addition, medium conditioned by continually paced myocytes contained significantly more bFGF measured by ELISA and more mitogenic activity for 3T3 cells, activity that could be reduced by a neutralizing anti-bFGF antibody. The hypothesis that transient membrane disruptions sufficient to allow release of cytosolic bFGF occur in paced myocytes was examined by monitoring the rate of uptake into myocytes from the medium of 10-kD dextran linked to fluorescein. Paced myocytes exhibited a significantly higher rate of fluorescein-labeled dextran uptake. These data are consistent with the hypothesis that nonlethal, transient alterations in sarcolemmal membrane permeability with release of cytosolic bFGF is one mechanism by which increased mechanical activity could lead to a hypertrophic response in cardiac myocytes.

Original languageEnglish (US)
Pages (from-to)281-291
Number of pages11
JournalJournal of Clinical Investigation
Volume97
Issue number2
DOIs
StatePublished - Jan 15 1996

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Fibroblast Growth Factor 2
Muscle Cells
Permeability
Membranes
Mechanical Stress
Neutralizing Antibodies
Cardiac Myocytes
Anti-Idiotypic Antibodies
3T3 Cells
Serum-Free Culture Media
Conditioned Culture Medium
Growth
Dextrans
Fluorescein
Phenylalanine
Cell Size
In Vitro Techniques
Protein Isoforms
Enzyme-Linked Immunosorbent Assay

Keywords

  • 2,3-butanedione monoxime
  • Angiogenesis
  • Fluorescence-activated cell sorting

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Role of transiently altered sarcolemmal membrane permeability and basic fibroblast growth factor release in the hypertrophic response of adult rat ventricular myocytes to increased mechanical activity in vitro. / Kaye, David; Pimental, David; Prasad, Sanjay; Mäki, Tiina; Berger, Hans Jörg; McNeil, Paul L.; Smith, Thomas W.; Kelly, Ralph A.

In: Journal of Clinical Investigation, Vol. 97, No. 2, 15.01.1996, p. 281-291.

Research output: Contribution to journalArticle

Kaye, David ; Pimental, David ; Prasad, Sanjay ; Mäki, Tiina ; Berger, Hans Jörg ; McNeil, Paul L. ; Smith, Thomas W. ; Kelly, Ralph A. / Role of transiently altered sarcolemmal membrane permeability and basic fibroblast growth factor release in the hypertrophic response of adult rat ventricular myocytes to increased mechanical activity in vitro. In: Journal of Clinical Investigation. 1996 ; Vol. 97, No. 2. pp. 281-291.
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