Putative mechanism of antiepileptic action of Vigabatrin on the pyramidal cells in rat neocortical slices studied by patch-clamp recording

Cellular electrophysiological properties are normally monitored by standard patch clamp techniques . The combination of membrane potential dyes with time-lapse laser confocal microscopy provides a more direct, least destructive rapid method for monitoring changes in neuronal electrical activity. Using membrane potential dyes we found that spontaneous action potential firing can be detected using time-lapse confocal microscopy. Initially, patch clamp recording techniques were used to verify spontaneous electrical activity in GH4\C1 pituitary cells. It was found that serum depleted cells had reduced spontaneous electrical activity. Brief exposure to the serum derived growth factor, IGF-1, reconstituted electrical activity. We have examined the possibility of developing a rapid fluorescent assay to measure neuronal activity using membrane potential dyes. This neuronal regeneration assay has been adapted to run on a confocal microscope. Quantitative fluorescence is then used to measure a compounds ability to regenerate neuronal firing.The membrane potential dye di-8-ANEPPS was selected for these experiments. Di-8- ANEPPS is internalized slowly, has a high signal to noise ratio (40:1), has a linear fluorescent response to change in voltage.

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Error analysis of Cm measurement under the whole-cell patch-clamp recording

Journal of Neuroscience Methods ◽

10.1016/j.jneumeth.2009.10.003 ◽

2010 ◽

Vol 185 (2) ◽

pp. 307-314 ◽

Cited By ~ 5

Author(s):

Hao Zhang ◽

Anlian Qu ◽

Jie Luo ◽

Jie Luo

Keyword(s):

Patch Clamp ◽

Error Analysis ◽

Patch Clamp Recording ◽

Whole Cell ◽

Cell Patch Clamp ◽

Whole Cell Patch Clamp

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Patch-Clamp Recording from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Improving Action Potential Characteristics through Dynamic Clamp

International Journal of Molecular Sciences ◽

10.3390/ijms18091873 ◽

2017 ◽

Vol 18 (9) ◽

pp. 1873 ◽

Cited By ~ 23

Author(s):

Arie Verkerk ◽

Christiaan Veerman ◽

Jan Zegers ◽

Isabella Mengarelli ◽

Connie Bezzina ◽

...

Keyword(s):

Stem Cell ◽

Patch Clamp ◽

Action Potential ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Dynamic Clamp ◽

Patch Clamp Recording ◽

Induced Pluripotent

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Electrophysiological Properties of Substantia Gelatinosa Neurons in the Preparation of a Slice of Middle-Aged Rat Spinal Cord

Frontiers in Aging Neuroscience ◽

10.3389/fnagi.2021.640265 ◽

2021 ◽

Vol 13 ◽

Author(s):

Yang Li ◽

Shanchu Su ◽

Jiaqi Yu ◽

Minjing Peng ◽

Shengjun Wan ◽

...

Keyword(s):

Spinal Cord ◽

Patch Clamp ◽

Molecular Mechanisms ◽

Substantia Gelatinosa ◽

Membrane Properties ◽

Middle Aged ◽

Patch Clamp Recording ◽

Electrophysiological Properties ◽

Glutamatergic Neurons ◽

Single Spike

A patch-clamp recording in slices generated from the brain or the spinal cord has facilitated the exploration of neuronal circuits and the molecular mechanisms underlying neurological disorders. However, the rodents that are used to generate the spinal cord slices in previous studies involving a patch-clamp recording have been limited to those in the juvenile or adolescent stage. Here, we applied an N-methyl-D-glucamine HCl (NMDG-HCl) solution that enabled the patch-clamp recordings to be performed on the superficial dorsal horn neurons in the slices derived from middle-aged rats. The success rate of stable recordings from substantia gelatinosa (SG) neurons was 34.6% (90/260). When stimulated with long current pulses, 43.3% (39/90) of the neurons presented a tonic-firing pattern, which was considered to represent γ-aminobutyric acid-ergic (GABAergic) signals. Presumptive glutamatergic neurons presented 38.9% (35/90) delayed and 8.3% (7/90) single-spike patterns. The intrinsic membrane properties of both the neuron types were similar but delayed (glutamatergic) neurons appeared to be more excitable as indicated by the decreased latency and rheobase values of the action potential compared with those of tonic (GABAergic) neurons. Furthermore, the glutamatergic neurons were integrated, which receive more excitatory synaptic transmission. We demonstrated that the NMDG-HCl cutting solution could be used to prepare the spinal cord slices of middle-aged rodents for the patch-clamp recording. In combination with other techniques, this preparation method might permit the further study of the functions of the spinal cord in the pathological processes that occur in aging-associated diseases.

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