title: In situ patch-clamp recordings from Merkel cells in rat whisker hair follicles, an experimental protocol for studying tactile transduction in tactile-end organs.
abstract: Mammals use tactile end-organs to perform sensory tasks such as environmental exploration, social interaction, and tactile discrimination. However, cellular and molecular mechanisms underlying tactile transduction in tactile end-organs remain poorly understood. The patch-clamp recording technique may be the most valuable approach for detecting and studying tactile transduction in tactile end-organs, but it is technically challenging because tactile transduction elements in an end-organ are normally inaccessible by patch-clamp recording electrodes. Here we describe an in situ patch-clamp recording protocol for the study of tactile transduction in Merkel cells of rat whisker hair follicles, one of the most sensitive tactile end-organs in mammals. This technique offers an opportunity to explore the identities and properties of ion channels that are involved in tactile transduction in whisker hair follicles, and it may also lend a useful tool for researchers to study other tactile end-organs. The experimental protocol describes procedures for 1) tissue dissection and whisker hair follicle preparation, 2) device setup and steps for performing patch-clamp recordings from Merkel cells in a whisker hair follicle, 3) methods of delivering mechanical stimuli, and 4) intra-follicle microinjection for receptor knockdown in whisker hair follicles. The main procedures in this protocol, from tissue preparation to whole-cell patch-clamp recordings, can be completed in a few hours.
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, College of Medicine, University of Alabama at Birmingham, 901 19TH Street South, BMR II 210, Birmingham, AL, 35294, USA. kneeikeda@gmail.com.|Department of Orthopaedic Surgery, Jikei University School of Medicine, 3-25-8 Nishi-Shinbashi, Minato-ku, Tokyo, 105-8461, Japan. kneeikeda@gmail.com.
authors.forename: Ryo
authors.initials: R
authors.lastname: Ikeda
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, College of Medicine, University of Alabama at Birmingham, 901 19TH Street South, BMR II 210, Birmingham, AL, 35294, USA. Jling64@uab.edu.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Ling
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, College of Medicine, University of Alabama at Birmingham, 901 19TH Street South, BMR II 210, Birmingham, AL, 35294, USA. myeounghoon.cha@gmail.com.
authors.forename: Myeounghoon
authors.initials: M
authors.lastname: Cha
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, College of Medicine, University of Alabama at Birmingham, 901 19TH Street South, BMR II 210, Birmingham, AL, 35294, USA. gujo@uab.edu.
authors.forename: Jianguo G
authors.initials: JG
authors.lastname: Gu
mesh_terms: D000818:Animals; D018859:Hair Follicle / Q000502:physiology*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D018862:Merkel Cells / Q000502:physiology*; D018408:Patch-Clamp Techniques; D017207:Rats, Sprague-Dawley; D014110:Touch / Q000502:physiology*; D014738:Vibrissae / Q000502:physiology*