Abstract:
:The synthetic cRNA encoding for the major T lymphocyte K+ channel (Kv1.3) was injected into Xenopus fertilized eggs. Somites from embryos of stage 20-22 (about 40 h post-fertilization at 19 degrees C) were dissociated and myotomal muscle cells were cultured in vitro for 2 days. The whole cell configuration of the tight seal patch-clamp technique was used to record K+ channel activity in cultured myocytes. These myocytes have two endogenous delayed-rectifiers (sustained and transient) and an inward-rectifier K+ currents, all of which are insensitive to the scorpion toxin charybdotoxin. Cultured myocytes dissociated from embryos injected with the Kv1.3 cRNA expressed the exogenous Kv1.3 channel. The Kv1.3 channel was identified by its physiological (a very low recovery from inactivation) and its pharmacological properties (a high sensitivity to charybdotoxin). This work demonstrates that Xenopus cultured myotomal muscle cells represent a very efficient and practical assay system for the functional expression of cloned ion channels.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Honoré E,Guillemare E,Lesage F,Barhanin J,Lazdunski Mdoi
10.1016/0014-5793(94)00622-9subject
Has Abstractpub_date
1994-07-18 00:00:00pages
259-62issue
3eissn
0014-5793issn
1873-3468pii
0014-5793(94)00622-9journal_volume
348pub_type
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