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Journal of Neuroscience, Vol 16, 456-466, Copyright © 1996 by Society for Neuroscience
Fractional calcium current through neuronal AMPA-receptor channels with a low calcium permeability
F Tempia, M Kano, R Schneggenburger, C Schirra, O Garaschuk, T Plant and A Konnerth
I. Physiologisches Institut, Universitat des Saarlandes, Homburg, Germany.
The Ca(2+)-permeation properties of AMPA-receptor (AMPA-R) channels in
Purkinje neurons in rat cerebellar slices were studied using a combination
of whole-cell patch-clamp recordings, Fura-2 fluorometry, and single-cell
reverse-transcription (RT)-PCR. Several lines of evidence indicate that
Purkinje neurons, at both early and late stages of postnatal development,
express exclusively AMPA-R channels with a low Ca2+ permeability. First, no
Ca2+ signal was detected during application of either AMPA or kainate to
Purkinje neurons loaded with the Ca2+ indicator Fura-2 AM. In contrast,
kainate application induced large Ca2+ transients in Bergmann glia cells.
Second, in ion substitution experiments, when Ca2+ is the only
extracellular permeant cation, the reversal potential corresponds to that
expected for AMPA-R channels with a low permeability for Ca2+. Third, using
a fluorometric flux-measurement approach (Schneggenburger et al., 1993a),
we found that the Ca2+ fraction of the total cation current through AMPA-R
channels is approximately 0.6%. This value is approximately sixfold lower
than that found for recombinant AMPA-R lacking the AMPA-R subunit GluR2.
Furthermore, single-cell RT-PCR experiments revealed the presence of the
AMPA-R subunits GluR1, GluR2, and GluR3 in Purkinje neurons in cerebellar
slices at developmental stages corresponding to those studied
electrophysiologically. The expression of GluR2 in all cells tested (n =
14) is consistent with the subunit composition predicted from studies of
recombinant AMPA-R channels with a low permeability for Ca2+ (Burnashev et
al., 1992b). In conclusion, this study establishes that cerebellar Purkinje
neurons at all postnatal developmental stages possess AMPA-R channels with
a low permeability for Ca2+.
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