Phosphothioated oligodeoxynucleotides induce nonspecific effects on neuronal cell adhesion in a growth substrate-dependent manner.
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.
Okun
Eitan
E
Laboratory of Neuroscience, National Institute on Aging, Intramural Research Program, Baltimore, Maryland 21224, USA. okune@mail.nih.gov
Mattson
Mark P
MP
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D016477:Artifacts; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D003627:Data Interpretation, Statistical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D010641:Phenotype; D010766:Phosphorylation; D011094:Polyethyleneimine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D051217:Toll-Like Receptor 9 / Q000187:drug effects / Q000378:metabolism
D000590:Amino Acid Chloromethyl Ketones; D003470:Culture Media; D004791:Enzyme Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D009838:Oligodeoxyribonucleotides; C489640:Tlr9 protein, rat; D051217:Toll-Like Receptor 9; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D011094:Polyethyleneimine; D053148:Caspase 3