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Volumn 284, Issue 5421, 1999, Pages 1811-1816

Rapid spine delivery and redistribution of AMPA receptors after synaptic NMDA receptor activation

Author keywords

[No Author keywords available]

Indexed keywords

AMPA RECEPTOR; GLUTAMATE RECEPTOR; GREEN FLUORESCENT PROTEIN; N METHYL DEXTRO ASPARTIC ACID RECEPTOR; RECEPTOR SUBUNIT;

EID: 0033546052     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.284.5421.1811     Document Type: Article
Times cited : (1083)

References (55)
  • 2
    • 0003457678 scopus 로고
    • D. Muller and G. Lynch, Proc. Natl. Acad. Sci. U.S.A. 85, 9346 (1988); J. A. Kauer et al., Neuron 1, 911 (1988); S. N. Davies et al., Nature 338, 500 (1989).
    • (1988) Proc. Natl. Acad. Sci. U.S.A. , vol.85 , pp. 9346
    • Muller, D.1    Lynch, G.2
  • 3
    • 0024151163 scopus 로고
    • D. Muller and G. Lynch, Proc. Natl. Acad. Sci. U.S.A. 85, 9346 (1988); J. A. Kauer et al., Neuron 1, 911 (1988); S. N. Davies et al., Nature 338, 500 (1989).
    • (1988) Neuron , vol.1 , pp. 911
    • Kauer, J.A.1
  • 4
    • 0024584295 scopus 로고
    • D. Muller and G. Lynch, Proc. Natl. Acad. Sci. U.S.A. 85, 9346 (1988); J. A. Kauer et al., Neuron 1, 911 (1988); S. N. Davies et al., Nature 338, 500 (1989).
    • (1989) Nature , vol.338 , pp. 500
    • Davies, S.N.1
  • 5
    • 0029018512 scopus 로고
    • D. Liao et al., Nature 375, 400 (1995); J. T. R. Isaac et al., Neuron 15, 427 (1995); G. Durand et al., Nature 381, 71 (1996); Y. Ben-Ari et al., Trends Neurosci. 20, 523 (1997).
    • (1995) Nature , vol.375 , pp. 400
    • Liao, D.1
  • 6
    • 0029162513 scopus 로고
    • D. Liao et al., Nature 375, 400 (1995); J. T. R. Isaac et al., Neuron 15, 427 (1995); G. Durand et al., Nature 381, 71 (1996); Y. Ben-Ari et al., Trends Neurosci. 20, 523 (1997).
    • (1995) Neuron , vol.15 , pp. 427
    • Isaac, J.T.R.1
  • 7
    • 0029921194 scopus 로고    scopus 로고
    • D. Liao et al., Nature 375, 400 (1995); J. T. R. Isaac et al., Neuron 15, 427 (1995); G. Durand et al., Nature 381, 71 (1996); Y. Ben-Ari et al., Trends Neurosci. 20, 523 (1997).
    • (1996) Nature , vol.381 , pp. 71
    • Durand, G.1
  • 8
    • 0030613643 scopus 로고    scopus 로고
    • D. Liao et al., Nature 375, 400 (1995); J. T. R. Isaac et al., Neuron 15, 427 (1995); G. Durand et al., Nature 381, 71 (1996); Y. Ben-Ari et al., Trends Neurosci. 20, 523 (1997).
    • (1997) Trends Neurosci. , vol.20 , pp. 523
    • Ben-Ari, Y.1
  • 9
    • 0032565873 scopus 로고    scopus 로고
    • T. A. Benke et al., Nature 393, 793 (1998).
    • (1998) Nature , vol.393 , pp. 793
    • Benke, T.A.1
  • 10
    • 0030175899 scopus 로고    scopus 로고
    • K. W. Roche et al., Neuron 16, 1179 (1996); Y. Hayashi et al., Mol. Brain Res. 46, 338 (1997); A. Barria et al., Science 276, 2042 (1997); V. Derkach et al., Proc. Natl. Acad. Sci. U.S.A. 96, 3269 (1999).
    • (1996) , vol.16 , pp. 1179
    • Roche, K.W.1
  • 11
    • 0030986005 scopus 로고    scopus 로고
    • K. W. Roche et al., Neuron 16, 1179 (1996); Y. Hayashi et al., Mol. Brain Res. 46, 338 (1997); A. Barria et al., Science 276, 2042 (1997); V. Derkach et al., Proc. Natl. Acad. Sci. U.S.A. 96, 3269 (1999).
    • (1997) Mol. Brain Res. , vol.46 , pp. 338
    • Hayashi, Y.1
  • 12
    • 0030744875 scopus 로고    scopus 로고
    • K. W. Roche et al., Neuron 16, 1179 (1996); Y. Hayashi et al., Mol. Brain Res. 46, 338 (1997); A. Barria et al., Science 276, 2042 (1997); V. Derkach et al., Proc. Natl. Acad. Sci. U.S.A. 96, 3269 (1999).
    • (1997) Science , vol.276 , pp. 2042
    • Barria, A.1
  • 13
    • 0032588030 scopus 로고    scopus 로고
    • K. W. Roche et al., Neuron 16, 1179 (1996); Y. Hayashi et al., Mol. Brain Res. 46, 338 (1997); A. Barria et al., Science 276, 2042 (1997); V. Derkach et al., Proc. Natl. Acad. Sci. U.S.A. 96, 3269 (1999).
    • (1999) Proc. Natl. Acad. Sci. U.S.A. , vol.96 , pp. 3269
    • Derkach, V.1
  • 17
    • 0345550786 scopus 로고    scopus 로고
    • note
    • GFP (enhanced GFP; Clontech) was inserted between the third and fourth amino acids after the predicted signal peptide cleavage site of rat GluR1-flop cDNA with standard molecular biology techniques. HEK cells were transfected with plasmid-based mammalian expression vector with lipofectin (Gibco-BRL Life Technologies). GluR1-GFP and GluR2 were cotransfected in 1:4 ratio. Protein immunoblotting was carried out with antibodies to GluR1 COOH-terminal (0.1 μg/ml; Chemicon International). Whole-cell recordings were obtained 2 to 5 days after transfection in Hepes (10 mM)-buffered Hanks' solution in the presence of 100 μM cyclothiazide. Kainate (1 mM) was applied through a puffer pipette positioned close to recorded cell Five to ten records were obtained at holding potentials of -80 to 60 mV (20-mV steps).
  • 18
    • 0029961358 scopus 로고    scopus 로고
    • I. Frolov et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11371 (1996); R. Malinow et al., in Imaging Living Cells (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, in press).
    • (1996) Proc. Natl. Acad. Sci. U.S.A. , vol.93 , pp. 11371
    • Frolov, I.1
  • 19
    • 0029961358 scopus 로고    scopus 로고
    • Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, in press
    • I. Frolov et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11371 (1996); R. Malinow et al., in Imaging Living Cells (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, in press).
    • Imaging Living Cells
    • Malinow, R.1
  • 20
    • 0344688658 scopus 로고    scopus 로고
    • unpublished results
    • Whole-cell recordings from neurons infected with Sindbis virus for 1 to 4 days show normal passive membrane properties (for example, input resistance: uninfected, 276 ± 64, N = 13; GluR1-GFP infected, 302 ± 51, N = 8; P = 0.78) (S. Shi, Y. Hayashi, R. Malinow, unpublished results).
    • Shi, S.1    Hayashi, Y.2    Malinow, R.3
  • 21
    • 0344688657 scopus 로고    scopus 로고
    • note
    • Dissociated cultured neurons were prepared as previously described (33).
  • 22
    • 0345550785 scopus 로고    scopus 로고
    • note
    • To estimate the level of recombinant GluR1-GFP expression, relative to endogenous GluR1 expression, we performed immunohistochemistry on fixed (14) dissociated hippocampal neurons (33) with a GluR1 COOH-terminal antibody (1 μg/ml; Chemicon International) that recognizes both proteins (Fig. 1B) as primary and Texas Red coupled as secondary (which does not overlap in fluorescence with GFP). In a field of infected and noninfected cells, dendrites (80 μm from cell body) of infected cells showed 2.7 ± 0.2 fold (mean ± SEM, N = 8) immunolabel compared with similar regions of uninfected cells.
  • 23
    • 0345119458 scopus 로고    scopus 로고
    • note
    • Tissue was fixed with freshly made 4% paraformaldehyde and 4% sucrose in phosphate-buffered saline (PBS) (dissociated neurons: 4 °C for 30 min; organotypic slices (76): 4 °C for 1 hour). With such fixation, immunohistochemistry (see below) detected only surface epitopes (dissociated cells: Fig. 2C; organotypic slices: Fig. 3G). To detect intracellular epitopes, tissue was further treated with 0.3% Triton X-100 in PBS (dissociated cells: 4 °C for 10 min; organotypic slices: 4 °C for 30 min). Immunohistochemistry: Cells were blocked in blocking solution (10% horse serum in PBS; 60 min) and then incubated with primary antibody (4 °C, overnight) in blocking solution. All primary rabbit polyclonal antibodies were visualized with biotin-conjugated antibody to rabbit immunoglobulin G (IgG) and Texas Red-avidin system. Mouse monoclonal antibodies were visualized with antibodies to mouse IgG labeled with Cy5 (Jackson ImmuoResearch). Images of dissociated neurons were collected with a cooled charge-coupled device (Photometrics) and analyzed with PMIS or V for Windows (Photometries); organotypic slices were imaged with TPLSM (21). Fraction of GluR1-GFP molecules on surface for a given region can be quantified by computing [(antibody to GFP signal)/(GFP signal)] in nonpermeabilized conditions for that region and normalizing by [(antibody to GFP signal)/(GFP signal)] obtained from similar tissue in permeabilized conditions. Signals in above computation indicate background subtracted values.
  • 24
    • 0345119457 scopus 로고    scopus 로고
    • note
    • 2 at 28 °C. Responses to glutamate were evoked by adding (in μM) 40 L-glutamic acid γ-(α-carboxyl-2-nitrobenzyl)ester (Molecular Probes), 1 tetrodotoxin, 100 picrotoxin, 100 APV, and 1 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphoric acid. Ten to 25 ms of flash (100 W of Hg; quartz optics, Zeiss Axiovert 135) was delivered to a spot (∼50-μm diameter) positioned over dendrites.
  • 25
    • 0025805120 scopus 로고
    • Organotypic slice culture of hippocampus was prepared as described [L. Stoppini et al., J. Neurosci. Methods 37, 173 (1991)]. Hippocampal slices (400 μm) were cut from postnatal 6-to 8-day-old rats with tissue chopper and cultured in medium as described [W. Musleh et al., Proc. Natl. Acad. Sci. U.S.A. 94, 9451 (1997)]. Slices were placed in a recording chamber and perfused with ACSF. Synaptic transmission was evoked (0.1 Hz, 100 μs, ∼10 V) with a glass stimulation electrode (1 to 2 megaohms, filled with 3 M NaCl), positioned in CA1 stratum radiatum. Tetanic stimuli consisted of two 100-Hz trains of 1-s duration separated by 20 s.
    • (1991) J. Neurosci. Methods , vol.37 , pp. 173
    • Stoppini, L.1
  • 26
    • 0030837569 scopus 로고    scopus 로고
    • Organotypic slice culture of hippocampus was prepared as described [L. Stoppini et al., J. Neurosci. Methods 37, 173 (1991)]. Hippocampal slices (400 μm) were cut from postnatal 6-to 8-day-old rats with tissue chopper and cultured in medium as described [W. Musleh et al., Proc. Natl. Acad. Sci. U.S.A. 94, 9451 (1997)]. Slices were placed in a recording chamber and perfused with ACSF. Synaptic transmission was evoked (0.1 Hz, 100 μs, ∼10 V) with a glass stimulation electrode (1 to 2 megaohms, filled with 3 M NaCl), positioned in CA1 stratum radiatum. Tetanic stimuli consisted of two 100-Hz trains of 1-s duration separated by 20 s.
    • (1997) Proc. Natl. Acad. Sci. U.S.A. , vol.94 , pp. 9451
    • Musleh, W.1
  • 27
    • 0028579743 scopus 로고
    • D. L. Pettit et al., Science 266, 1881 (1994).
    • (1994) Science , vol.266 , pp. 1881
    • Pettit, D.L.1
  • 29
    • 0030610646 scopus 로고    scopus 로고
    • A. Miyawaki et al., Nature 388, 882 (1997).
    • (1997) Nature , vol.388 , pp. 882
    • Miyawaki, A.1
  • 30
    • 0345550784 scopus 로고    scopus 로고
    • note
    • The distribution of GluR1 and GluR1-GFP was measured quantitatively with immuno-gold labeling (18). Sections from dendritic regions ∼100 μm from the cell bodies were examined. To identify a cell infected with GluR1-GFP virus, we identified a dendritic segment (∼1 μm in diameter) with immunolabeling. Comparable regions (with dendritic segments ∼1 μm in diameter) were chosen in tissue from uninfected P10 animals for analysis of endogenous GluR1. For each section, the label was identified as being in one of the following compartments: A, dendritic cytosol; B, dendritic surface; C, spine, non-PSD; and D, PSD. The total number of grains counted was 515 for endogenous GluR1 and 785 for GluR1-GFP. For background subtraction, 50 presynaptic terminals were randomly chosen, label was counted, and density was calculated (0.17 gold per square micrometer). This background density constituted 10 to 20% of the signal found in dendritic cytosol (0.92 gold per square micrometer) and was subtracted to reach dendritic cytosol value (A, above); this region is the only compartment with sufficient area to be affected by background labeling.
  • 31
    • 0030023673 scopus 로고    scopus 로고
    • K. Svoboda et al., Science 272, 716 (1996); W. Denk and K. Svoboda, Neuron 18, 351 (1997); Z. F. Mainen et al., Methods, in press. Optical stacks (30 μm by 30 μm by 50 μm; step size. 0.5 μm) were captured about every 15 min with a custom-built two-photon laser scanning microscope [Zeiss, 63X objective; Ti: sapphire laser tuned to wavelength (λ) ∼ 900 nm]. Stimulation electrode was placed nearby dendrites expressing GluR1-GFP under visual guidance. Quantification of fluorescence from images was carried out with custom-made programs in IDL (Research Systems). Images were imported into Adobe Photoshop for figure presentation. Each displayed image is generally the average of three to five optical sections. In some cases, an image is the composite of images captured at slightly different (∼1 μm) optical sections; this was necessary to offset tilting of structures over the course of hours of examination.
    • (1996) Science , vol.272 , pp. 716
    • Svoboda, K.1
  • 32
    • 0030993430 scopus 로고    scopus 로고
    • K. Svoboda et al., Science 272, 716 (1996); W. Denk and K. Svoboda, Neuron 18, 351 (1997); Z. F. Mainen et al., Methods, in press. Optical stacks (30 μm by 30 μm by 50 μm; step size. 0.5 μm) were captured about every 15 min with a custom-built two-photon laser scanning microscope [Zeiss, 63X objective; Ti: sapphire laser tuned to wavelength (λ) ∼ 900 nm]. Stimulation electrode was placed nearby dendrites expressing GluR1-GFP under visual guidance. Quantification of fluorescence from images was carried out with custom-made programs in IDL (Research Systems). Images were imported into Adobe Photoshop for figure presentation. Each displayed image is generally the average of three to five optical sections. In some cases, an image is the composite of images captured at slightly different (∼1 μm) optical sections; this was necessary to offset tilting of structures over the course of hours of examination.
    • (1997) Neuron , vol.18 , pp. 351
    • Denk, W.1    Svoboda, K.2
  • 33
    • 0030023673 scopus 로고    scopus 로고
    • in press
    • K. Svoboda et al., Science 272, 716 (1996); W. Denk and K. Svoboda, Neuron 18, 351 (1997); Z. F. Mainen et al., Methods, in press. Optical stacks (30 μm by 30 μm by 50 μm; step size. 0.5 μm) were captured about every 15 min with a custom-built two-photon laser scanning microscope [Zeiss, 63X objective; Ti: sapphire laser tuned to wavelength (λ) ∼ 900 nm]. Stimulation electrode was placed nearby dendrites expressing GluR1-GFP under visual guidance. Quantification of fluorescence from images was carried out with custom-made programs in IDL (Research Systems). Images were imported into Adobe Photoshop for figure presentation. Each displayed image is generally the average of three to five optical sections. In some cases, an image is the composite of images captured at slightly different (∼1 μm) optical sections; this was necessary to offset tilting of structures over the course of hours of examination.
    • Methods
    • Mainen, Z.F.1
  • 34
    • 0345550782 scopus 로고    scopus 로고
    • note
    • 2+ imaging studies with the same stimulus electrodes (tip resistance, 1 to 2 megaohms) and parameters indicate that synaptic excitation in this preparation is sparse. Thus, as in previous studies (21, 37), the tip of the stimulating electrode must be placed around 5 to 15 μm from imaged postsynaptic regions.
  • 35
    • 0345550783 scopus 로고    scopus 로고
    • note
    • Within 3 SD of the noise measured in background regions.
  • 36
    • 0345550780 scopus 로고    scopus 로고
    • note
    • -1, mean ± SD, measured over 30 randomly chosen dendritic segments).
  • 37
    • 0344688653 scopus 로고    scopus 로고
    • note
    • 50%) as an index of signal homogeneity.
  • 38
    • 0345119456 scopus 로고    scopus 로고
    • note
    • This decrease is 3.6 SD beyond those changes seen in the absence of stimulation, indicating a small likelihood that these changes occur from chance alone.
  • 42
    • 0032536033 scopus 로고    scopus 로고
    • P.-M. Lledo et al., ibid. 279, 399 (1998).
    • (1998) Science , vol.279 , pp. 399
    • Lledo, P.-M.1
  • 43
    • 0032127472 scopus 로고    scopus 로고
    • A. Nishimune et al., Neuron 21, 87 (1998); P. Osten et al., ibid., p. 99; I. Song et al., ibid., p. 393.
    • (1998) Neuron , vol.21 , pp. 87
    • Nishimune, A.1
  • 44
    • 0032127472 scopus 로고    scopus 로고
    • A. Nishimune et al., Neuron 21, 87 (1998); P. Osten et al., ibid., p. 99; I. Song et al., ibid., p. 393.
    • Neuron , pp. 99
    • Osten, P.1
  • 45
    • 0032127472 scopus 로고    scopus 로고
    • A. Nishimune et al., Neuron 21, 87 (1998); P. Osten et al., ibid., p. 99; I. Song et al., ibid., p. 393.
    • Neuron , pp. 393
    • Song, I.1
  • 49
    • 0032167980 scopus 로고    scopus 로고
    • I. Nusser et al., Neuron 21, 545 (1998).
    • (1998) Neuron , vol.21 , pp. 545
    • Nusser, I.1
  • 51
    • 0032214532 scopus 로고    scopus 로고
    • P. Worley, Neuron 21, 936 (1998).
    • (1998) Neuron , vol.21 , pp. 936
    • Worley, P.1
  • 52
    • 25344458686 scopus 로고
    • R. Malinow et al., Proc. Natl. Acad. Sci. U.S.A. 91, B170 (1994); R. Yuste and W. Denk, Nature 375, 682 (1995); H. Takechi et al., ibid. 396, 757 (1998).
    • (1994) , vol.91
    • Malinow, R.1
  • 53
    • 0029065159 scopus 로고
    • R. Malinow et al., Proc. Natl. Acad. Sci. U.S.A. 91, B170 (1994); R. Yuste and W. Denk, Nature 375, 682 (1995); H. Takechi et al., ibid. 396, 757 (1998).
    • (1995) Nature , vol.375 , pp. 682
    • Yuste, R.1    Denk, W.2
  • 54
    • 0032585513 scopus 로고    scopus 로고
    • R. Malinow et al., Proc. Natl. Acad. Sci. U.S.A. 91, B170 (1994); R. Yuste and W. Denk, Nature 375, 682 (1995); H. Takechi et al., ibid. 396, 757 (1998).
    • (1998) Nature , vol.396 , pp. 757
    • Takechi, H.1
  • 55
    • 0344688648 scopus 로고    scopus 로고
    • note
    • We thank S. Schlesinger for Sindbis virus expression system; P. De Camilli for antibody against synapsin I; C. S. Zucker for antibody to GFP; J. Boulter, S. F. Heinemann, and P. H. Seeburg for cDNA clones; N. Dawkins-Pisani, B. Burbach, and P. O'Brian for technical assistance; and Y.-X. Wang for assistance in the immunogold studies. Y.H. is a recipient of research fellowships from Japan Society for the Promotion of Science and Uehara Memorial Foundation. S.H.Z. was a recipient of Wellcome Trust Fellowship. This study was supported by the NIH (R.M. and K.S.), the Mathers Foundation (R.M.), and the Human Frontier Science Program, Pew, and Whitaker Foundations (K.S).


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