Journal article
Gramicidin-perforated patch recording: GABA response in mammalian neurones with intact intracellular chloride
The Journal of physiology, Vol.484(Pt 1), pp.77-86
04/01/1995
DOI: 10.1113/jphysiol.1995.sp020649
PMCID: PMC1157923
PMID: 7541464
Abstract
1. By the development of a new perforated patch method using gramicidin, the effects of GABA on neurones dissociated from the rat substantia nigra pars reticulata (SNR) were examined without disturbing the intracellular chloride concentration. 2. Using the patch pipette solution containing gramicidin (100 micrograms ml-1), the access resistance dropped to less than 20 M omega within 40 min after making the gigaohm seal. 3. Under current-clamp conditions, GABA caused a hyperpolarization accompanied by a blockade of spontaneous firing. Under voltage clamp at a holding potential (Vh) of -50 mV, GABA evoked an outward current by way of bicuculline- and picrotoxin-sensitive GABAA receptors. 4. A 10-fold change of extracellular chloride concentration resulted in a 58 mV shift of the reversal potential of GABA-induced outward current (EGABA), indicating that the membrane behaves like a chloride electrode in the presence of GABA. 5. The intracellular chloride activities (aCli), calculated with the Nernst equation using both extracellular chloride activity and EGABA values, ranged from 2.8 to 19.7 mM with a mean value of 9.5 mM. The aCli was not affected either by different pipette solutions or by different holding potentials more hyperpolarized than -40 mV. 6. In the recording from SNR neurones in brain slice using the gramicidin-perforated patch-clamp technique, the inhibitory and excitatory postsynaptic currents were recorded in different current directions and the former was blocked by bicuculline. 7. In conclusion, the gramicidin-perforated patch method will disclose previously unknown aspects of biological responses involving Cl-.
Details
- Title: Subtitle
- Gramicidin-perforated patch recording: GABA response in mammalian neurones with intact intracellular chloride
- Creators
- S Ebihara - Department of Physiology, Kyushu University Faculty of Medicine, Fukuoka, JapanK Shirato - Department of Physiology, Kyushu University Faculty of Medicine, Fukuoka, JapanN Harata - Department of Physiology, Kyushu University Faculty of Medicine, Fukuoka, JapanN Akaike - Department of Physiology, Kyushu University Faculty of Medicine, Fukuoka, Japan
- Resource Type
- Journal article
- Publication Details
- The Journal of physiology, Vol.484(Pt 1), pp.77-86
- DOI
- 10.1113/jphysiol.1995.sp020649
- PMID
- 7541464
- PMCID
- PMC1157923
- ISSN
- 0022-3751
- eISSN
- 1469-7793
- Language
- English
- Date published
- 04/01/1995
- Academic Unit
- Molecular Physiology and Biophysics
- Record Identifier
- 9984025598502771
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