AMPA receptor subunits expressed by single purkinje cells

Lambolez, Bertrand; Audinat, Etienne; Bochet, Pascal; Crépel, F.; Rossier, Jean

doi:10.1016/0896-6273(92)90164-9

Cited by 514 publications

(343 citation statements)

References 25 publications

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“…For single-cell RT-PCR analysis (Lambolez et al, 1992;Monyer and Jonas, 1995) of mGluR1 and mGluR5 expression, the cytoplasm of interneurons was aspirated into patch pipettes, pulled from autoclaved boroscilicate glass (resistance of 1.5-2.5 M⍀, filled with autoclaved internal solution), by gentle suction under visual control after the electrophysiological recordings had been made. The harvested material was subsequently expelled into an autoclaved tube containing desoxyribonucleoside triphosphates, dithiothreitol, ribonuclease inhibitor, and Superscript reverse transcriptase, and incubated for 1 hr at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Differential Expression of Group I Metabotropic Glutamate Receptors in Functionally Distinct Hippocampal Interneurons

et al. 2000

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Metabotropic glutamate receptors (mGluRs) have been proposed to be involved in oscillatory rhythmic activity in the hippocampus. However, the subtypes of mGluRs involved and their precise distribution in different populations of interneurons is unclear. In this study, we combined functional analysis of mGluR-mediated inward currents in CA1 oriens-alveus interneurons with anatomical and immunocytochemical identification of these interneurons and expression analysis of group I mGluR using single-cell reverse transcription-PCR (RT-PCR). Four major interneuron subtypes could be distinguished based on the mGluR-mediated inward current induced by the application of 100 M trans-(1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) under voltage-clamp conditions and the action potential firing pattern under current-clamp conditions. Type I interneurons responded with a large inward current of ϳ224 pA, were positive for somatostatin, and the majority expressed both mGluR1 and mGluR5. Type II interneurons responded with an inward current of ϳ80 pA, contained calbindin, and expressed mainly mGluR1. Type III interneurons responded with an inward current of ϳ60 pA. These interneurons were fast-spiking, contained parvalbumin, and expressed mainly mGluR5. Type IV interneurons did not respond with an inward current upon application of ACPD, yet they expressed group I mGluRs. Activation of group I mGluRs under currentclamp conditions increased spike frequency and resulted in rhythmic firing activity in type I and II, but not in type III and IV, interneurons. RT-PCR results suggest that activation of mGluR1 in the subsets of GABAergic interneurons, classified here as type I and II, may play an important role in mediating synchronous activity.

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Section: Methodsmentioning

confidence: 99%

Differential Expression of Group I Metabotropic Glutamate Receptors in Functionally Distinct Hippocampal Interneurons

et al. 2000

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“…A fine microcapillary is employed to extract cell sap from the individual cells, after which RT-PCR is performed (Lambolez et al 1992;Karrer et al1995;Brandt et al 1999;Koroleva et al 2001). The critical advantage of single-cell (SC)-RT-PCR is that it enables expression to be measured directly from living tissue at the finest spatial resolution possible, that of the individual cell.…”

Section: Introductionmentioning

confidence: 99%

“…The critical advantage of single-cell (SC)-RT-PCR is that it enables expression to be measured directly from living tissue at the finest spatial resolution possible, that of the individual cell. Moreover, because the microcapillary can also be used to measure, in single cells, parameters such as membrane potential, ionic status, metabolite contents and wall mechanical properties (Lambolez et al 1992;Tomos and Leigh 1999), changes in these can be linked in turn to gene expression (Koroleva et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Distribution of actin gene isoforms in the Arabidopsis leaf measured in microsamples from intact individual cells

Laval

Koroleva

Murphy

et al. 2002

Planta

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The contents of single plant cells can be sampled using glass microcapillaries. By combining such single-cell sampling with reverse transcription-polymerase chain reaction (RT-PCR), transcripts of individual genes can be identified and, in principle, quantified. This provides a valuable technique for the analysis and quantification of the intercellular distribution of gene expression in complex tissues. In a proof-of-principle study, the cellular locations of the transcripts of the eight isoforms of actin (ACT) expressed in Arabidopsis thaliana (L.) Heynh. were analyzed. Cell sap was extracted from epidermal and mesophyll cells of leaves of 3-to 4-week-old plants. Single-cell (SC)-RT-PCR was used to amplify the actin transcripts using specific primer pairs for ACT1, 2, 3, 4, 7, 8, 11 and 12. Only ACT2 and ACT8 were found in epidermal and in mesophyll cells. In individual trichomes, in addition to ACT2 and ACT8, ACT7 and ACT11 transcripts were detectable. By employing the already well-characterized actin system we demonstrate the practicality and power of SC-RT-PCR as a technique for analyzing gene expression at the ultimate level of resolution, the single cell.

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“…Studies of native AM PARs by patch-clamp recordings combined with single-cell reverse transcription-PCR (single-cell RT-PCR; Lambolez et al, 1992) have confirmed the role played by the GluR2 subunit in their permeation (Bochet et al, 1994;Jonas et al, 1994;Geiger et al, 1995), but the identity of the molecular determinants controlling their kinetics is still controversial. Indeed, slow and fast desensitizations were correlated with either GluR2 flip and GluR4 (Geiger et al, 1995), respectively, or to flip and flop splice variants (Lambolez et al, 1996).…”

Section: Ampa Receptor; Neocortex; Interneuron; Single-cell Rt-pcr; Fmentioning

confidence: 99%

“…In heterologous expression systems recombinant AM PAR permeation properties are controlled by the relative abundance of GluR2 (for review on voltage dependence and calcium permeability, see Hollmann and Heinemann, 1994;Jonas and Burnashev, 1995) [see also Swanson et al (1997) for single channel conductance]. In contrast, their kinetic properties are affected by multiple factors, including subunit composition, flip/flop alternative splicing, and mRNA edition at the R/G site (Sommer et al, 1990;L omeli et al, 1994;Mosbacher et al, 1994;Partin et al, 1994).Studies of native AM PARs by patch-clamp recordings combined with single-cell reverse transcription-PCR (single-cell RT-PCR; Lambolez et al, 1992) have confirmed the role played by the GluR2 subunit in their permeation (Bochet et al, 1994;Jonas et al, 1994;Geiger et al, 1995), but the identity of the molecular determinants controlling their kinetics is still controversial. Indeed, slow and fast desensitizations were correlated with either GluR2 flip and GluR4 (Geiger et al, 1995), respectively, or to flip and flop splice variants (Lambolez et al, 1996).…”

mentioning

confidence: 99%

Subunit Composition, Kinetic, and Permeation Properties of AMPA Receptors in Single Neocortical Nonpyramidal Cells

Angulo¹,

Lambolez²,

Audinat³

et al. 1997

J. Neurosci.

Self Cite

123

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Native AMPA receptors (AMPARs) were investigated in neocortical fast-spiking (FS) and regular-spiking nonpyramidal (RSNP) cells. The onset of and recovery from desensitization as well as current rectification and single-channel conductance were studied by using fast glutamate application to outside-out patches. The GluR1-4 subunit, flip/flop splicing, and R/G editing expression patterns of functionally characterized cells were determined by single-cell reverse transcription-PCR to correlate the subunit composition of native AMPARs with their functional properties. Our sample, mostly constituted by RSNP neurons, predominantly expressed GluR3 flip and GluR2 flop. In individual cells, flip/flop splicing of each subunit appeared to be regulated independently, whereas for R/G editing all subunits were either almost fully edited or unedited. We confirmed that the relative GluR2 expression controls the permeation properties of native AMPARs, whereas none of the single molecular parameters considered appeared to be a key determinant of the kinetics. FS neurons displayed AMPARs with relatively homogeneous functional properties characterized by fast desensitization, slow recovery from desensitization, marked inward rectification, and large single-channel conductance. In contrast, these parameters varied over a wide range in RSNP neurons, and their combination resulted in various AMPAR functional patterns. Indeed, in different cells, fast or slow desensitization was found to be associated with either slow or fast recovery from desensitization. Similarly, fast or slow kinetics was associated with either strong or weak rectification. Our results suggest that kinetic and permeation properties of native AMPARs can be regulated independently in cortical neurons and probably do not have the same molecular determinants. AMPA receptor; neocortex; interneuron; single-cell RT-PCR; fast glutamate application; subunit; splicing; editingThe fast excitatory synaptic transmission in the C NS is mediated mainly by AM PA receptor channels (AM PARs). These receptors are multimeric assemblies of four different subunits GluR1-4 (for review, see Wisden and Seeburg, 1993;Hollmann and Heinemann, 1994). Further diversity is generated by alternative splicing (Sommer et al., 1990) and mRNA editing (Sommer et al., 1991;L omeli et al., 1994). In heterologous expression systems recombinant AM PAR permeation properties are controlled by the relative abundance of GluR2 (for review on voltage dependence and calcium permeability, see Hollmann and Heinemann, 1994;Jonas and Burnashev, 1995) [see also Swanson et al. (1997) for single channel conductance]. In contrast, their kinetic properties are affected by multiple factors, including subunit composition, flip/flop alternative splicing, and mRNA edition at the R/G site (Sommer et al., 1990;L omeli et al., 1994;Mosbacher et al., 1994;Partin et al., 1994).Studies of native AM PARs by patch-clamp recordings combined with single-cell reverse transcription-PCR (single-cell RT-PCR; Lambolez et al., 1992) have con...

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AMPA receptor subunits expressed by single purkinje cells

Cited by 514 publications

References 25 publications

Differential Expression of Group I Metabotropic Glutamate Receptors in Functionally Distinct Hippocampal Interneurons

Differential Expression of Group I Metabotropic Glutamate Receptors in Functionally Distinct Hippocampal Interneurons

Distribution of actin gene isoforms in the Arabidopsis leaf measured in microsamples from intact individual cells

Subunit Composition, Kinetic, and Permeation Properties of AMPA Receptors in Single Neocortical Nonpyramidal Cells

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