High-resolution iontophoresis for single-synapse stimulation.
We present a technique for stimulating post-synaptic receptors with neurotransmitter locally at a single synapse and with a concentration profile that is comparable to endogenous stimulation. We modify the technique of iontophoresis to use a 0.1 microm electrode tip for local stimulation, and we combine it with fast capacitance compensation to achieve high-speed application from a high-resistance tip. Ejection of fluorescent dye from the electrode shows that transmitter can be limited to the width of a single synapse and to a time scale similar to an endogenous event. The speed and localization of transmitter is confirmed by iontophoretically stimulating single labeled synapses in cultured hippocampal neurons held under voltage clamp. The amount of transmitter ejected is linear and reproducible over a physiologically relevant range, making this technique useful for examining receptor kinetics and receptor insertion/removal. The system should be capable of delivering any charged neurotransmitter, and we show examples using glutamate and GABA. The technique is also combined with computer-controlled manipulation to study the strength and plasticity of multiple synapses in real-time.
Murnick
Jonathan G
JG
Department of Biology, RIKEN-MIT Neuroscience Research Center, Center for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Dubé
Gilles
G
Krupa
Boris
B
Liu
Guosong
G
D000818:Animals; D002264:Carboxylic Acids / Q000493:pharmacokinetics; D002478:Cells, Cultured; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D018698:Glutamic Acid / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D007478:Iontophoresis / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D051381:Rats; D018091:Receptors, AMPA / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000502:physiology; D013569:Synapses / Q000502:physiology*; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D002264:Carboxylic Acids; D005456:Fluorescent Dyes; C436864:Oregon Green 488 carboxylic acid; D018091:Receptors, AMPA; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid