Room temperature culture extends the useful life of adult neurons for voltage-clamp experiments.
Isolated superior cervical ganglion (SCG) neurons were maintained in vitro at 22 degrees C (22 degrees C neurons) for up to 4 days in an effort to inhibit process outgrowth and thus extend the useful life of SCG neurons for voltage-clamp experiments. The neurons were viable after 4 days in vitro and remained roughly spherical whereas neurons maintained in vitro at 37 degrees C (37 degrees C neurons) developed extensive neurite processes after 2 days. The resting potential of 22 degrees C neurons was more hyperpolarized and the action potential duration was reduced compared to acutely isolated neurons (acute neurons) or 37 degrees C neurons. The steady state Na+ current activation and inactivation parameters of the acute and 22 degrees C neurons were similar whereas the half activation voltage was hyperpolarized and the slope factor of inactivation was reduced for the 37 degrees C neurons compared to the other two groups. The Na+ currents recorded from the 37 degrees C neurons displayed obvious signs of poor voltage control which were not observed in the acute or 22 degrees C neurons. The acetylcholine (ACh) sensitivity of both 22 degrees C and 37 degrees C neurons was significantly less than that of the acute neurons. This report demonstrates that room temperature culture of SCG neurons is a simple method which prevents process outgrowth and thus extends the useful life of the neurons for voltage-clamp experiments.
Magee
J C
JC
Department of Physiology, Tulane University Medical School, New Orleans, LA 70112.
Schofield
G G
GG
D000109:Acetylcholine / Q000494:pharmacology; D000200:Action Potentials; D002470:Cell Survival; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods*; D005728:Ganglia, Sympathetic / Q000166:cytology; D015640:Ion Channel Gating / Q000187:drug effects; D008564:Membrane Potentials; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D012964:Sodium / Q000378:metabolism; D013696:Temperature
D012964:Sodium; D000109:Acetylcholine