Polyethylenimine-mediated DNA transfection of peripheral and central neurons in primary culture: probing Ca2+ channel structure and function with antisense oligonucleotides.
To study neuronal ion channel function with antisense oligonucleotides, a reliable method is needed which allows different neuronal cell types to be transfected without artifactual disruptive effects on their electrical properties. Here we report that use of the recently introduced transfecting agent, polyethylenimine, fulfills this requirement. Four days after transfection, in both central and peripheral neurons, an antisense designed to block the synthesis of the Ca2+ channel beta subunits induced a maximal decrease of the Ca2- current amplitude and modification of their kinetics and voltage-dependence. Controls with scrambled oligonucleotides, as well as Na+ current recordings of antisense transfected neurons, confirm both that the transfecting agent does not modify the electrophysiological properties of the neurons and that the effect of the antisense is sequence specific.
Lambert
R C
RC
Laboratoire de Neurobiologie Cellulaire, CNRS (UPR 9009), Strasbourg, France.
Maulet
Y
Y
Dupont
J L
JL
Mykita
S
S
Craig
P
P
Volsen
S
S
Feltz
A
A
D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000737:chemistry / Q000235:genetics*; D002478:Cells, Cultured / Q000737:chemistry / Q000502:physiology; D004247:DNA / Q000235:genetics; D004594:Electrophysiology; D009474:Neurons / Q000737:chemistry / Q000166:cytology*; D016376:Oligonucleotides, Antisense; D010525:Peripheral Nerves / Q000166:cytology*; D011094:Polyethyleneimine / Q000494:pharmacology*; D051381:Rats; D014162:Transfection / Q000379:methods*
D015220:Calcium Channels; D016376:Oligonucleotides, Antisense; D011094:Polyethyleneimine; D004247:DNA