A procedure for localisation and electrophysiological characterisation of ion channels heterologously expressed in a plant context
Articolo
Data di Pubblicazione:
2005
Citazione:
A procedure for localisation and electrophysiological
characterisation of ion channels heterologously expressed in a plant
context / E. Hosy, G. Duby, A.A. Véry, A. Costa, H. Sentenac, J.B. Thibaud. - In: PLANT METHODS. - ISSN 1746-4811. - 1:1(2005 Dec), pp. 14.14-14.14.
Abstract:
Background: In silico analyses based on sequence similarities with animal channels have identified
a large number of plant genes likely to encode ion channels. The attempts made to characterise
such putative plant channels at the functional level have most often relied on electrophysiological
analyses in classical expression systems, such as Xenopus oocytes or mammalian cells. In a number
of cases, these expression systems have failed so far to provide functional data and one can
speculate that using a plant expression system instead of an animal one might provide a more
efficient way towards functional characterisation of plant channels, and a more realistic context to
investigate regulation of plant channels.
Results: With the aim of developing a plant expression system readily amenable to
electrophysiological analyses, we optimised experimental conditions for preparation and
transformation of tobacco mesophyll protoplasts and engineered expression plasmids, that were
designed to allow subcellular localisation and functional characterisation of ion channels eventually
in presence of their putative (possibly over-expressed) regulatory partners. Two inward K+
channels from the Shaker family were functionally expressed in this system: not only the compliant
KAT1 but also the recalcitrant AKT1 channel, which remains electrically silent when expressed in
Xenopus oocytes or in mammalian cells.
Conclusion: The level of endogenous currents in control protoplasts seems compatible with the
use of the described experimental procedures for the characterisation of plant ion channels, by
studying for instance their subcellular localisation, functional properties, structure-function
relationships, interacting partners and regulation, very likely in a more realistic context than the
classically used animal systems
Tipologia IRIS:
01 - Articolo su periodico
Elenco autori:
E. Hosy, G. Duby, A.A. Véry, A. Costa, H. Sentenac, J.B. Thibaud
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