Domestic application of infrared patch clamp techniques on brain slices is limited.The key of the tech-nique is to prepare high-quality brain slices.The present paper describes the preparation procedure of brainstem s...Domestic application of infrared patch clamp techniques on brain slices is limited.The key of the tech-nique is to prepare high-quality brain slices.The present paper describes the preparation procedure of brainstem slices and the spontaneous firing properties of rat medial vestibular nucleus(MVN)neurons.By infrared differ-ential interference contrast technique,neurons of rat MVN were visualized directly at the depth of 50–100 mm underneath the surface of slices.Firing activities of MVN neurons were recorded by the whole-cell patch clamp technique in artificial cerebrospinal fluid(ACSF)and low Ca^(2+)-high Mg^(2+) fluid.The firing mode was more irregular and depressive in low Ca^(2+)-high Mg2+fluid than in ACSF.According to the averaged waveform of action potentials,cells were classified as the neurons with mono-phasic after-hyperpolarization potential(AHP),and the neurons with biphasic AHP.The resting membrane potential(RMP),input resistance(Rin)and membrane capacitance(Cm)of neurons were recorded and com-pared between groups.With infrared videomicroscopy,patch clamp recordings could be made under direct obser-vation in freshly prepared brainstem slices.The discharge activities of MVN neurons were spontaneous and the fir-ing mode was modulated by extracellular calcium concen-tration.The basic membrane properties of two types of neurons were not significantly different,while the differ-ences in waveform might play a role in the segregation between tonic and kinetic cells.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.30371525)the National Science Foundation for Distinguished Young Scholars of China(No.39925035)the National Science&Technology Pillar Program in the Eleventh Five-year Plan Period(No.2007BAI18B13).
文摘Domestic application of infrared patch clamp techniques on brain slices is limited.The key of the tech-nique is to prepare high-quality brain slices.The present paper describes the preparation procedure of brainstem slices and the spontaneous firing properties of rat medial vestibular nucleus(MVN)neurons.By infrared differ-ential interference contrast technique,neurons of rat MVN were visualized directly at the depth of 50–100 mm underneath the surface of slices.Firing activities of MVN neurons were recorded by the whole-cell patch clamp technique in artificial cerebrospinal fluid(ACSF)and low Ca^(2+)-high Mg^(2+) fluid.The firing mode was more irregular and depressive in low Ca^(2+)-high Mg2+fluid than in ACSF.According to the averaged waveform of action potentials,cells were classified as the neurons with mono-phasic after-hyperpolarization potential(AHP),and the neurons with biphasic AHP.The resting membrane potential(RMP),input resistance(Rin)and membrane capacitance(Cm)of neurons were recorded and com-pared between groups.With infrared videomicroscopy,patch clamp recordings could be made under direct obser-vation in freshly prepared brainstem slices.The discharge activities of MVN neurons were spontaneous and the fir-ing mode was modulated by extracellular calcium concen-tration.The basic membrane properties of two types of neurons were not significantly different,while the differ-ences in waveform might play a role in the segregation between tonic and kinetic cells.