Perfusion system components release agents that distort functional properties of rod cyclic nucleotide-gated ion channels.
In switching from studying native cyclic nucleotide-gated (CNG) ion channels in rod cells to studying the corresponding cloned channels expressed in Xenopus oocytes, we changed our perfusion system to a more efficient one. This change involved replacing culture flasks and a small plexiglass/glass chamber with plastic syringes, metal needles, and plastic petri dishes. We now report that these new perfusion system components release agents that distort or obscure measured functional properties of rod CNG channels. The magnitude and time course of appearance of the artifacts vary widely among individual components (e.g. from syringe to syringe). The effects most resemble voltage-dependent block of the channels, giving a decrease in current at positive potentials, and producing distortions of the kinetics and voltage dependence of channel activation.
Crary
J I
JI
Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, RI 02912, USA.
Gordon
S E
SE
Zimmerman
A L
AL
D000818:Animals; D016477:Artifacts; D006152:Cyclic GMP / Q000494:pharmacology*; D004594:Electrophysiology; D005260:Female; D015640:Ion Channel Gating / Q000502:physiology*; D007473:Ion Channels / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D009339:Needles; D009865:Oocytes; D010477:Perfusion / Q000295:instrumentation / Q000379:methods*; D017948:Retinal Rod Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism*; D013594:Syringes; D014982:Xenopus laevis
D007473:Ion Channels; D006152:Cyclic GMP