Probing localized neural mechanotransduction through surface-modified elastomeric matrices and electrophysiology
Autor: | Chao-Min Cheng, Yi-Wen Lin, Philip R. LeDuc, Chih-Cheng Chen, Robert L. Steward, Yuan-Ren Cheng, Robert M. Bellin |
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Rok vydání: | 2010 |
Předmět: |
Patch-Clamp Techniques
Materials science Sensory Receptor Cells Neurite Models Neurological Context (language use) Stimulation Sensory system Anatomy In Vitro Techniques Mechanotransduction Cellular General Biochemistry Genetics and Molecular Biology Electrophysiological Phenomena Mice Electrophysiology Mechanobiology Coated Materials Biocompatible Elastomers Neurites Biophysics Animals Dimethylpolysiloxanes Patch clamp Mechanotransduction |
Zdroj: | Nature Protocols. 5:714-724 |
ISSN: | 1750-2799 1754-2189 |
DOI: | 10.1038/nprot.2010.15 |
Popis: | Mechanotransduction of sensory neurons is of great interest to the scientific community, especially in areas such as pain, neurobiology, cardiovascular homeostasis and mechanobiology. We describe a method to investigate stretch-activated mechanotransduction in sensory nerves through subcellular stimulation. The method imposes localized mechanical stimulation through indentation of an elastomeric substrate and combines this mechanical stimulation with whole-cell patch clamp recording of the electrical response to single-nerve stretching. One significant advantage here is that the neurites are stretched with limited physical contact beyond their attachment to the polymer. When we imposed specific mechanical stimulation through the substrate, the stretched neurite fired and an action potential response was recorded. In addition, complementary protocols to control the molecules at the cell-substrate interface are presented. These techniques provide an opportunity to probe neurosensory mechanotransduction with a defined substrate, whose physical and molecular context can be modified to mimic physiologically relevant conditions. The entire process from fabrication to cellular recording takes 5 to 6 d. |
Databáze: | OpenAIRE |
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