Probing localized neural mechanotransduction through surface-modified elastomeric matrices and electrophysiology.
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.
Cheng
Chao-Min
CM
Department of Mechanical and Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.
Lin
Yi-Wen
YW
Bellin
Robert M
RM
Steward
Robert L
RL
Cheng
Yuan-Ren
YR
LeDuc
Philip R
PR
Chen
Chih-Cheng
CC
D000818:Animals; D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes; D020724:Elastomers; D055724:Electrophysiological Phenomena; D066298:In Vitro Techniques; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D051379:Mice; D008959:Models, Neurological; D016501:Neurites / Q000502:physiology; D018408:Patch-Clamp Techniques; D011984:Sensory Receptor Cells / Q000502:physiology*
D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes; D020724:Elastomers; C013830:baysilon