[Gramicidin perforated patch recording technique].
Cl- is one of the major ionic constituents of cells and extracellular spaces. Intracellular Cl- plays an important role in regulating the cell volume and pH, in both salt secretion and reabsorption, in membrane excitability, and G-protein-dependent intracellular signal transduction. GABA and glycine are the primary inhibitory neurotransmitters. Such agonist-stimulated responses are affected by the intracellular Cl- concentration ([Cl-]i). However, it was difficult to make an electrical recording of the physiological Cl- response from cells with native intracellular Cl- activity because of the limitations of present recording techniques using conventional glass-microelectrode, whole-cell patch and nystatin perforated patch recording modes. Recently, this difficulty was overcome by developing the gramicidin perforated patch recording mode in our laboratory. Gramicidin is a polypeptide antibiotic that forms pores in the cell membrane as well as nystatin but allows only monovalent cations to permeate the membrane, enabling both [Cl-]i and the second messenger system to remain undisturbed. Here, I would like to primarily focus on the GABA- and glycine-induced Cl- responses to in mammalian CNS neurons maintaining native cellular [Cl-]i under normal and pathological neuronal conditions by use of gramicidin perforated patch recording configuration. Age-related and developmental changes in neuronal [Cl-]i are also described in detail.
Akaike
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Department of Physiology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
D000818:Animals; D000900:Anti-Bacterial Agents; D001693:Biological Transport, Active; D001921:Brain / Q000378:metabolism; D018118:Chloride Channels / Q000378:metabolism; D002713:Chlorine / Q000378:metabolism; D005998:Glycine / Q000502:physiology; D006096:Gramicidin; D009474:Neurons / Q000378:metabolism; D018408:Patch-Clamp Techniques; D005680:gamma-Aminobutyric Acid / Q000502:physiology
D000900:Anti-Bacterial Agents; D018118:Chloride Channels; D006096:Gramicidin; D002713:Chlorine; D005680:gamma-Aminobutyric Acid; D005998:Glycine