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Random sequence phosphorothioate oligonucleotides evoke dramatic phenotypic alterations in cardiac myocyte cultures
D L Davis1, I C Doten, K F Barald
1Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
Antisense & Nucleic Acid Drug Development
|January 1, 1996
Summary
Phosphorothioate oligonucleotides (oligos) surprisingly promote cardiac myocyte aggregation and coordinated contraction. These findings suggest potential applications for phosphorothioate oligos in cardiac cell culture and therapy.
Area of Science:
- Cardiology
- Molecular Biology
- Cell Biology
Background:
- Cardiac myocyte culture is essential for research but can lead to loss of function.
- Unmodified oligonucleotides (oligos) have no significant effect on myocyte behavior.
- Phosphorothioate (PS) oligos are chemically modified nucleic acid sequences.
Purpose of the Study:
- To investigate the effect of random sequence phosphorothioate (PS) oligonucleotides on cardiac myocyte behavior in vitro.
- To determine if PS oligos can improve the functional properties of cultured cardiac myocytes.
- To explore the potential of PS oligos as cell culture reagents for cardiac myocytes.
Main Methods:
- Culturing cardiac myocytes with and without unmodified or PS oligos.
- Assessing myocyte contractile activity and organization.
- Measuring gap junction conductivity using photobleaching.
- Analyzing atrial natriuretic factor (ANF) gene expression.
Main Results:
- Cardiac myocytes cultured with PS oligos formed cable-like aggregates with coordinated contractile activity.
- Gap junction conductivity increased fourfold in myocytes treated with PS oligos.
- PS oligos repressed aberrant atrial natriuretic factor (ANF) gene expression observed in control cultures.
- Unmodified oligos did not induce these effects.
Conclusions:
- PS oligos significantly enhance the organization and function of cultured cardiac myocytes.
- PS oligos may mitigate negative effects of myocyte isolation and culturing.
- PS oligos show promise as valuable reagents for cardiac myocyte cell culture.
- The potential in vivo effects of PS oligos in intact hearts warrant further investigation for clinical significance.
