Preparation of Acute Spinal Cord Slices for Whole-cell Patch-clamp Recording in Substantia Gelatinosa Neurons

10.3791/58479 ◽  
2019 ◽  
Author(s):  
Mengye Zhu ◽  
Daying Zhang ◽  
Sicong Peng ◽  
Nana Liu ◽  
Jing Wu ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Patch Clamp ◽  
Substantia Gelatinosa ◽  
Patch Clamp Recording ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp

Whole Cell Recordings From Visualized Neurons in the Inner Laminae of the Functionally Intact Spinal Cord

2009 ◽  
Vol 102 (1) ◽  
pp. 590-597 ◽  
Author(s):  
Jason Dyck ◽  
Simon Gosgnach
Keyword(s):  
Neural Network ◽  
Spinal Cord ◽  
Locomotor Activity ◽  
Patch Clamp ◽  
Lumbar Spinal Cord ◽  
Whole Cell ◽  
Neuronal Populations ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp

The in vitro whole spinal cord preparation has been an invaluable tool for the study of the neural network that underlies walking because it provides a means of recording fictive locomotor activity following surgical and/or pharmacological manipulation. The recent use of molecular genetic techniques to identify discrete neuronal populations in the spinal cord and subsequent studies showing some of these populations to be involved in locomotor activity have been exciting developments that may lead to a better understanding of the structure and mechanism of function of this neural network. It would be of great benefit if the in vitro whole spinal cord preparation could be updated to allow for the direct targeting of genetically defined neuronal populations, allowing each to be characterized physiologically and anatomically. This report describes a new technique that enables the visualization of, and targeted whole cell patch-clamp recordings from, genetically defined populations of neurons while leaving connectivity largely intact. The key feature of this technique is a small notch cut in the lumbar spinal cord that reveals cells located in the intermediate laminae while leaving the ventral portion of the spinal cord—the region containing the locomotor neural network—untouched. Whole cell patch-clamp recordings demonstrate that these neurons are healthy and display large rhythmic depolarizations that are related to electroneurogram bursts recorded from ventral roots during fictive locomotion. Intracellular labeling demonstrates that this technique can also be used to map axonal projection patterns of neurons. We expect that this procedure will greatly facilitate electrophysiological and anatomical study of important neuronal populations that constitute neural networks throughout the CNS.

Z f-and-H sys-based C m measurement under the whole-cell patch-clamp recording

2008 ◽  
Vol 457 (6) ◽  
pp. 1423-1434 ◽  
Author(s):  
Hao Zhang ◽  
Jie Luo ◽  
Jun Xiong ◽  
Xian-Guang Lin ◽  
Zheng-Xing Wu ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Patch Clamp Recording ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp

GABA-Mimetic Actions of Withania somnifera on Substantia Gelatinosa Neurons of the Trigeminal Subnucleus Caudalis in Mice

2013 ◽  
Vol 41 (05) ◽  
pp. 1043-1051 ◽  
Author(s):  
Hua Yin ◽  
Dong Hyu Cho ◽  
Soo Joung Park ◽  
Seong Kyu Han
Keyword(s):  
Patch Clamp ◽  
Receptor Antagonist ◽  
Withania Somnifera ◽  
Substantia Gelatinosa ◽  
Patch Clamp Technique ◽  
Pipette Solution ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Inward Currents

The plant Withania somnifera (WS), also known as Ashwagandha, has been used widely in traditional medicine systems in India and Nepal (Ayurveda), and has been accepted to cure various ailments. In this study, the whole-cell patch clamp technique was performed to examine the mechanism of action of WS on the SG neurons of the Vc from mouse brainstem slices. In whole-cell patch clamp mode, methanol extract of Withania somnifera (mWS) induced short-lived and repeatable inward currents in all SG neurons tested (31.3±8.51 pA, n = 7) using a high chloride pipette solution. The mWS-induced inward currents were concentration dependent and maintained in the presence of tetrodotoxin (TTX), a voltage gated Na + channel blocker, CNQX, a non-NMDA glutamate receptor antagonist, AP5, an NMDA receptor antagonist and strychnine, a glycine receptor antagonist. The mWS induced currents were blocked by picrotoxin, a GABAA receptor antagonist. These results show that mWS has an inhibitory effects on SG neurons of the Vc through GABAA receptor-mediated activation of chloride ion channels, indicating that mWS contains compounds with sedative effects on the central nervous system. These results also suggest that mWS may be a potential target for modulating orofacial pain processing.

scholarly journals Whole Cell Patch Clamp Recording Performed on a Planar Glass Chip

2002 ◽  
Vol 82 (6) ◽  
pp. 3056-3062 ◽  
Author(s):  
Niels Fertig ◽  
Robert H. Blick ◽  
Jan C. Behrends
Keyword(s):  
Patch Clamp ◽  
Patch Clamp Recording ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Glass Chip
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