Phosphothioated oligodeoxynucleotides induce nonspecific effects on neuronal cell adhesion in a growth substrate-dependent manner

J Neurosci Res. 2009 Jun;87(8):1947-52. doi: 10.1002/jnr.21995.

Abstract

Synthetic phosphothioated (PTO) oligodeoxynucleotide (ODN) sequences are commonly used for a variety of applications that benefit from nuclease protection. The PTO modification is implemented mainly in antisense ODN, but also in ODN that were shown to activate members of the toll-like receptor (TLR) family such as TLR3 (poly-I:C), TLR8 (ssRNA), and TLR9 (CpG). Neurons are routinely plated on surfaces coated with either cationic substances such as poly-L-ornithine (PLO), polyethylenimine (PEI), poly-L-lysine or ECM components such as laminin, collagen, or fibronectin. We found that PTO-ODN aimed at activating TLR9 induces a non-TLR9-specific detachment phenotype in cortical neurons plated on either laminin or PEI, but not on PLO. This phenotype was correlated with decreased viability and was partially inhibited when caspase-3 was inhibited with Ac-DEVD-CMK. This finding suggests that the use of PTO-ODN can cause nonspecific effects on cell adhesion that could compromise interpretation of data from experiments using PTO-ODN.

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Animals
  • Artifacts
  • Caspase 3 / drug effects
  • Caspase 3 / metabolism
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Culture Techniques
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cells, Cultured
  • Culture Media / pharmacology*
  • Data Interpretation, Statistical
  • Enzyme Inhibitors / pharmacology
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Laminin / pharmacology
  • Neurons / cytology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Oligodeoxyribonucleotides / chemical synthesis
  • Oligodeoxyribonucleotides / chemistry
  • Oligodeoxyribonucleotides / pharmacology*
  • Phenotype
  • Phosphorylation
  • Polyethyleneimine / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Toll-Like Receptor 9 / drug effects
  • Toll-Like Receptor 9 / metabolism

Substances

  • Amino Acid Chloromethyl Ketones
  • Culture Media
  • Enzyme Inhibitors
  • Intercellular Signaling Peptides and Proteins
  • Laminin
  • Oligodeoxyribonucleotides
  • Tlr9 protein, rat
  • Toll-Like Receptor 9
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Polyethyleneimine
  • Caspase 3