TY - JOUR
T1 - Patch-clamp and perfusion techniques to study ion channels expressed in xenopus oocytes
AU - Zhang, Guohui
AU - Cui, Jianmin
N1 - Publisher Copyright:
© 2018 Cold Spring Harbor Laboratory Press.
PY - 2018/4
Y1 - 2018/4
N2 - The Xenopus oocyte expression system is ideal for electrophysiological characterization of voltage-dependent and ligand-dependent ion channels because of its relatively low background of endogenous channels and the large size of the cell. Here, we present a protocol to study voltage- and ligand-dependent activation of ion channels expressed in Xenopus oocytes using patch-clamp techniques designed to control both the membrane voltage and the intracellular solution. In this protocol, the large conductance voltage- and Ca2+ -activated K+ (BK) channel is studied as an example. After injection of BK channel mRNA, oocytes are incubated for 2–7 d at 18˚C. Inside-out membrane patches containing single or multiple BK channels are excised with perfusion of different solutions during recording. The protocol can be used to study structure–function relations for ion channels and neurotransmitter receptors.
AB - The Xenopus oocyte expression system is ideal for electrophysiological characterization of voltage-dependent and ligand-dependent ion channels because of its relatively low background of endogenous channels and the large size of the cell. Here, we present a protocol to study voltage- and ligand-dependent activation of ion channels expressed in Xenopus oocytes using patch-clamp techniques designed to control both the membrane voltage and the intracellular solution. In this protocol, the large conductance voltage- and Ca2+ -activated K+ (BK) channel is studied as an example. After injection of BK channel mRNA, oocytes are incubated for 2–7 d at 18˚C. Inside-out membrane patches containing single or multiple BK channels are excised with perfusion of different solutions during recording. The protocol can be used to study structure–function relations for ion channels and neurotransmitter receptors.
UR - http://www.scopus.com/inward/record.url?scp=85044941863&partnerID=8YFLogxK
U2 - 10.1101/pdb.prot099051
DO - 10.1101/pdb.prot099051
M3 - Article
C2 - 29382809
AN - SCOPUS:85044941863
SN - 1940-3402
VL - 2018
SP - 258
EP - 267
JO - Cold Spring Harbor Protocols
JF - Cold Spring Harbor Protocols
IS - 4
ER -