---
_id: '9756'
abstract:
- lang: eng
  text: High-resolution visualization and quantification of membrane proteins contribute
    to the understanding of their functions and the roles they play in physiological
    and pathological conditions. Sodium dodecyl sulfate-digested freeze-fracture replica
    labeling (SDS-FRL) is a powerful electron microscopy method to study quantitatively
    the two-dimensional distribution of transmembrane proteins and their tightly associated
    proteins. During treatment with SDS, intracellular organelles and proteins not
    anchored to the replica are dissolved, whereas integral membrane proteins captured
    and stabilized by carbon/platinum deposition remain on the replica. Their intra-
    and extracellular domains become exposed on the surface of the replica, facilitating
    the accessibility of antibodies and, therefore, providing higher labeling efficiency
    than those obtained with other immunoelectron microscopy techniques. In this chapter,
    we describe the protocols of SDS-FRL adapted for mammalian brain samples, and
    optimization of the SDS treatment to increase the labeling efficiency for quantification
    of Cav2.1, the alpha subunit of P/Q-type voltage-dependent calcium channels utilizing
    deep learning algorithms.
acknowledgement: This work was supported by the European Union (European Research
  Council Advanced grant no. 694539 and Human Brain Project Ref. 720270 to R. S.)
  and the Austrian Academy of Sciences (DOC fellowship to D.K.).
alternative_title:
- Neuromethods
article_processing_charge: No
author:
- first_name: Walter
  full_name: Kaufmann, Walter
  id: 3F99E422-F248-11E8-B48F-1D18A9856A87
  last_name: Kaufmann
  orcid: 0000-0001-9735-5315
- first_name: David
  full_name: Kleindienst, David
  id: 42E121A4-F248-11E8-B48F-1D18A9856A87
  last_name: Kleindienst
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
citation:
  ama: 'Kaufmann W, Kleindienst D, Harada H, Shigemoto R. High-Resolution localization
    and quantitation of membrane proteins by SDS-digested freeze-fracture replica
    labeling (SDS-FRL). In: <i> Receptor and Ion Channel Detection in the Brain</i>.
    Vol 169. Neuromethods. New York: Humana; 2021:267-283. doi:<a href="https://doi.org/10.1007/978-1-0716-1522-5_19">10.1007/978-1-0716-1522-5_19</a>'
  apa: 'Kaufmann, W., Kleindienst, D., Harada, H., &#38; Shigemoto, R. (2021). High-Resolution
    localization and quantitation of membrane proteins by SDS-digested freeze-fracture
    replica labeling (SDS-FRL). In <i> Receptor and Ion Channel Detection in the Brain</i>
    (Vol. 169, pp. 267–283). New York: Humana. <a href="https://doi.org/10.1007/978-1-0716-1522-5_19">https://doi.org/10.1007/978-1-0716-1522-5_19</a>'
  chicago: 'Kaufmann, Walter, David Kleindienst, Harumi Harada, and Ryuichi Shigemoto.
    “High-Resolution Localization and Quantitation of Membrane Proteins by SDS-Digested
    Freeze-Fracture Replica Labeling (SDS-FRL).” In <i> Receptor and Ion Channel Detection
    in the Brain</i>, 169:267–83. Neuromethods. New York: Humana, 2021. <a href="https://doi.org/10.1007/978-1-0716-1522-5_19">https://doi.org/10.1007/978-1-0716-1522-5_19</a>.'
  ieee: 'W. Kaufmann, D. Kleindienst, H. Harada, and R. Shigemoto, “High-Resolution
    localization and quantitation of membrane proteins by SDS-digested freeze-fracture
    replica labeling (SDS-FRL),” in <i> Receptor and Ion Channel Detection in the
    Brain</i>, vol. 169, New York: Humana, 2021, pp. 267–283.'
  ista: 'Kaufmann W, Kleindienst D, Harada H, Shigemoto R. 2021.High-Resolution localization
    and quantitation of membrane proteins by SDS-digested freeze-fracture replica
    labeling (SDS-FRL). In:  Receptor and Ion Channel Detection in the Brain. Neuromethods,
    vol. 169, 267–283.'
  mla: Kaufmann, Walter, et al. “High-Resolution Localization and Quantitation of
    Membrane Proteins by SDS-Digested Freeze-Fracture Replica Labeling (SDS-FRL).”
    <i> Receptor and Ion Channel Detection in the Brain</i>, vol. 169, Humana, 2021,
    pp. 267–83, doi:<a href="https://doi.org/10.1007/978-1-0716-1522-5_19">10.1007/978-1-0716-1522-5_19</a>.
  short: W. Kaufmann, D. Kleindienst, H. Harada, R. Shigemoto, in:,  Receptor and
    Ion Channel Detection in the Brain, Humana, New York, 2021, pp. 267–283.
date_created: 2021-07-30T09:34:56Z
date_published: 2021-07-27T00:00:00Z
date_updated: 2024-03-25T23:30:16Z
day: '27'
ddc:
- '573'
department:
- _id: RySh
- _id: EM-Fac
doi: 10.1007/978-1-0716-1522-5_19
ec_funded: 1
has_accepted_license: '1'
intvolume: '       169'
keyword:
- 'Freeze-fracture replica: Deep learning'
- Immunogold labeling
- Integral membrane protein
- Electron microscopy
language:
- iso: eng
month: '07'
oa_version: None
page: 267-283
place: New York
project:
- _id: 25CA28EA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '694539'
  name: 'In situ analysis of single channel subunit composition in neurons: physiological
    implication in synaptic plasticity and behaviour'
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
publication: ' Receptor and Ion Channel Detection in the Brain'
publication_identifier:
  eisbn:
  - '9781071615225'
  isbn:
  - '9781071615218'
publication_status: published
publisher: Humana
quality_controlled: '1'
related_material:
  record:
  - id: '9562'
    relation: dissertation_contains
    status: public
series_title: Neuromethods
status: public
title: High-Resolution localization and quantitation of membrane proteins by SDS-digested
  freeze-fracture replica labeling (SDS-FRL)
type: book_chapter
user_id: D865714E-FA4E-11E9-B85B-F5C5E5697425
volume: 169
year: '2021'
...
---
_id: '7391'
abstract:
- lang: eng
  text: Electron microscopy (EM) is a technology that enables visualization of single
    proteins at a nanometer resolution. However, current protein analysis by EM mainly
    relies on immunolabeling with gold-particle-conjugated antibodies, which is compromised
    by large size of antibody, precluding precise detection of protein location in
    biological samples. Here, we develop a specific chemical labeling method for EM
    detection of proteins at single-molecular level. Rational design of α-helical
    peptide tag and probe structure provided a complementary reaction pair that enabled
    specific cysteine conjugation of the tag. The developed chemical labeling with
    gold-nanoparticle-conjugated probe showed significantly higher labeling efficiency
    and detectability of high-density clusters of tag-fused G protein-coupled receptors
    in freeze-fracture replicas compared with immunogold labeling. Furthermore, in
    ultrathin sections, the spatial resolution of the chemical labeling was significantly
    higher than that of antibody-mediated labeling. These results demonstrate substantial
    advantages of the chemical labeling approach for single protein visualization
    by EM.
article_processing_charge: No
article_type: original
author:
- first_name: Shigekazu
  full_name: Tabata, Shigekazu
  id: 4427179E-F248-11E8-B48F-1D18A9856A87
  last_name: Tabata
- first_name: Marijo
  full_name: Jevtic, Marijo
  id: 4BE3BC94-F248-11E8-B48F-1D18A9856A87
  last_name: Jevtic
- first_name: Nobutaka
  full_name: Kurashige, Nobutaka
  last_name: Kurashige
- first_name: Hirokazu
  full_name: Fuchida, Hirokazu
  last_name: Fuchida
- first_name: Munetsugu
  full_name: Kido, Munetsugu
  last_name: Kido
- first_name: Kazushi
  full_name: Tani, Kazushi
  last_name: Tani
- first_name: Naoki
  full_name: Zenmyo, Naoki
  last_name: Zenmyo
- first_name: Shohei
  full_name: Uchinomiya, Shohei
  last_name: Uchinomiya
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Makoto
  full_name: Itakura, Makoto
  last_name: Itakura
- first_name: Itaru
  full_name: Hamachi, Itaru
  last_name: Hamachi
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Akio
  full_name: Ojida, Akio
  last_name: Ojida
citation:
  ama: Tabata S, Jevtic M, Kurashige N, et al. Electron microscopic detection of single
    membrane proteins by a specific chemical labeling. <i>iScience</i>. 2019;22(12):256-268.
    doi:<a href="https://doi.org/10.1016/j.isci.2019.11.025">10.1016/j.isci.2019.11.025</a>
  apa: Tabata, S., Jevtic, M., Kurashige, N., Fuchida, H., Kido, M., Tani, K., … Ojida,
    A. (2019). Electron microscopic detection of single membrane proteins by a specific
    chemical labeling. <i>IScience</i>. Elsevier. <a href="https://doi.org/10.1016/j.isci.2019.11.025">https://doi.org/10.1016/j.isci.2019.11.025</a>
  chicago: Tabata, Shigekazu, Marijo Jevtic, Nobutaka Kurashige, Hirokazu Fuchida,
    Munetsugu Kido, Kazushi Tani, Naoki Zenmyo, et al. “Electron Microscopic Detection
    of Single Membrane Proteins by a Specific Chemical Labeling.” <i>IScience</i>.
    Elsevier, 2019. <a href="https://doi.org/10.1016/j.isci.2019.11.025">https://doi.org/10.1016/j.isci.2019.11.025</a>.
  ieee: S. Tabata <i>et al.</i>, “Electron microscopic detection of single membrane
    proteins by a specific chemical labeling,” <i>iScience</i>, vol. 22, no. 12. Elsevier,
    pp. 256–268, 2019.
  ista: Tabata S, Jevtic M, Kurashige N, Fuchida H, Kido M, Tani K, Zenmyo N, Uchinomiya
    S, Harada H, Itakura M, Hamachi I, Shigemoto R, Ojida A. 2019. Electron microscopic
    detection of single membrane proteins by a specific chemical labeling. iScience.
    22(12), 256–268.
  mla: Tabata, Shigekazu, et al. “Electron Microscopic Detection of Single Membrane
    Proteins by a Specific Chemical Labeling.” <i>IScience</i>, vol. 22, no. 12, Elsevier,
    2019, pp. 256–68, doi:<a href="https://doi.org/10.1016/j.isci.2019.11.025">10.1016/j.isci.2019.11.025</a>.
  short: S. Tabata, M. Jevtic, N. Kurashige, H. Fuchida, M. Kido, K. Tani, N. Zenmyo,
    S. Uchinomiya, H. Harada, M. Itakura, I. Hamachi, R. Shigemoto, A. Ojida, IScience
    22 (2019) 256–268.
date_created: 2020-01-29T15:56:56Z
date_published: 2019-12-20T00:00:00Z
date_updated: 2024-03-25T23:30:07Z
day: '20'
ddc:
- '570'
department:
- _id: RySh
doi: 10.1016/j.isci.2019.11.025
ec_funded: 1
external_id:
  isi:
  - :000504652000020
  pmid:
  - '31786521'
file:
- access_level: open_access
  checksum: f3e90056a49f09b205b1c4f8c739ffd1
  content_type: application/pdf
  creator: dernst
  date_created: 2020-02-04T10:48:36Z
  date_updated: 2020-07-14T12:47:57Z
  file_id: '7448'
  file_name: 2019_iScience_Tabata.pdf
  file_size: 7197776
  relation: main_file
file_date_updated: 2020-07-14T12:47:57Z
has_accepted_license: '1'
intvolume: '        22'
issue: '12'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
page: 256-268
pmid: 1
project:
- _id: 25CA28EA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '694539'
  name: 'In situ analysis of single channel subunit composition in neurons: physiological
    implication in synaptic plasticity and behaviour'
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
publication: iScience
publication_identifier:
  issn:
  - 2589-0042
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  record:
  - id: '11393'
    relation: dissertation_contains
    status: public
scopus_import: '1'
status: public
title: Electron microscopic detection of single membrane proteins by a specific chemical
  labeling
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 22
year: '2019'
...
---
_id: '736'
abstract:
- lang: eng
  text: The neurotransmitter receptor subtype, number, density, and distribution relative
    to the location of transmitter release sites are key determinants of signal transmission.
    AMPA-type ionotropic glutamate receptors (AMPARs) containing GluA3 and GluA4 subunits
    are prominently expressed in subsets of neurons capable of firing action potentials
    at high frequencies, such as auditory relay neurons. The auditory nerve (AN) forms
    glutamatergic synapses on two types of relay neurons, bushy cells (BCs) and fusiform
    cells (FCs) of the cochlear nucleus. AN-BC and AN-FC synapses have distinct kinetics;
    thus, we investigated whether the number, density, and localization of GluA3 and
    GluA4 subunits in these synapses are differentially organized using quantitative
    freeze-fracture replica immunogold labeling. We identify a positive correlation
    between the number of AMPARs and the size of AN-BC and AN-FC synapses. Both types
    of AN synapses have similar numbers of AMPARs; however, the AN-BC have a higher
    density of AMPARs than AN-FC synapses, because the AN-BC synapses are smaller.
    A higher number and density of GluA3 subunits are observed at AN-BC synapses,
    whereas a higher number and density of GluA4 subunits are observed at AN-FC synapses.
    The intrasynaptic distribution of immunogold labeling revealed that AMPAR subunits,
    particularly GluA3, are concentrated at the center of the AN-BC synapses. The
    central distribution of AMPARs is absent in GluA3-knockout mice, and gold particles
    are evenly distributed along the postsynaptic density. GluA4 gold labeling was
    homogenously distributed along both synapse types. Thus, GluA3 and GluA4 subunits
    are distributed at AN synapses in a target-cell-dependent manner.
article_processing_charge: No
author:
- first_name: María
  full_name: Rubio, María
  last_name: Rubio
- first_name: Ko
  full_name: Matsui, Ko
  last_name: Matsui
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
- first_name: Naomi
  full_name: Kamasawa, Naomi
  last_name: Kamasawa
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Makoto
  full_name: Itakura, Makoto
  last_name: Itakura
- first_name: Elek
  full_name: Molnár, Elek
  last_name: Molnár
- first_name: Manabu
  full_name: Abe, Manabu
  last_name: Abe
- first_name: Kenji
  full_name: Sakimura, Kenji
  last_name: Sakimura
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
citation:
  ama: Rubio M, Matsui K, Fukazawa Y, et al. The number and distribution of AMPA receptor
    channels containing fast kinetic GluA3 and GluA4 subunits at auditory nerve synapses
    depend on the target cells. <i>Brain Structure and Function</i>. 2017;222(8):3375-3393.
    doi:<a href="https://doi.org/10.1007/s00429-017-1408-0">10.1007/s00429-017-1408-0</a>
  apa: Rubio, M., Matsui, K., Fukazawa, Y., Kamasawa, N., Harada, H., Itakura, M.,
    … Shigemoto, R. (2017). The number and distribution of AMPA receptor channels
    containing fast kinetic GluA3 and GluA4 subunits at auditory nerve synapses depend
    on the target cells. <i>Brain Structure and Function</i>. Springer. <a href="https://doi.org/10.1007/s00429-017-1408-0">https://doi.org/10.1007/s00429-017-1408-0</a>
  chicago: Rubio, María, Ko Matsui, Yugo Fukazawa, Naomi Kamasawa, Harumi Harada,
    Makoto Itakura, Elek Molnár, Manabu Abe, Kenji Sakimura, and Ryuichi Shigemoto.
    “The Number and Distribution of AMPA Receptor Channels Containing Fast Kinetic
    GluA3 and GluA4 Subunits at Auditory Nerve Synapses Depend on the Target Cells.”
    <i>Brain Structure and Function</i>. Springer, 2017. <a href="https://doi.org/10.1007/s00429-017-1408-0">https://doi.org/10.1007/s00429-017-1408-0</a>.
  ieee: M. Rubio <i>et al.</i>, “The number and distribution of AMPA receptor channels
    containing fast kinetic GluA3 and GluA4 subunits at auditory nerve synapses depend
    on the target cells,” <i>Brain Structure and Function</i>, vol. 222, no. 8. Springer,
    pp. 3375–3393, 2017.
  ista: Rubio M, Matsui K, Fukazawa Y, Kamasawa N, Harada H, Itakura M, Molnár E,
    Abe M, Sakimura K, Shigemoto R. 2017. The number and distribution of AMPA receptor
    channels containing fast kinetic GluA3 and GluA4 subunits at auditory nerve synapses
    depend on the target cells. Brain Structure and Function. 222(8), 3375–3393.
  mla: Rubio, María, et al. “The Number and Distribution of AMPA Receptor Channels
    Containing Fast Kinetic GluA3 and GluA4 Subunits at Auditory Nerve Synapses Depend
    on the Target Cells.” <i>Brain Structure and Function</i>, vol. 222, no. 8, Springer,
    2017, pp. 3375–93, doi:<a href="https://doi.org/10.1007/s00429-017-1408-0">10.1007/s00429-017-1408-0</a>.
  short: M. Rubio, K. Matsui, Y. Fukazawa, N. Kamasawa, H. Harada, M. Itakura, E.
    Molnár, M. Abe, K. Sakimura, R. Shigemoto, Brain Structure and Function 222 (2017)
    3375–3393.
date_created: 2018-12-11T11:48:14Z
date_published: 2017-11-01T00:00:00Z
date_updated: 2023-09-27T14:14:51Z
day: '01'
ddc:
- '571'
department:
- _id: RySh
doi: 10.1007/s00429-017-1408-0
external_id:
  isi:
  - '000414761700002'
file:
- access_level: open_access
  checksum: 73787a22507de8fb585bb598e1418ca7
  content_type: application/pdf
  creator: system
  date_created: 2018-12-12T10:10:20Z
  date_updated: 2020-07-14T12:47:56Z
  file_id: '4806'
  file_name: IST-2017-881-v1+1_s00429-017-1408-0.pdf
  file_size: 4011126
  relation: main_file
file_date_updated: 2020-07-14T12:47:56Z
has_accepted_license: '1'
intvolume: '       222'
isi: 1
issue: '8'
language:
- iso: eng
month: '11'
oa: 1
oa_version: Published Version
page: 3375 - 3393
publication: Brain Structure and Function
publication_identifier:
  issn:
  - '18632653'
publication_status: published
publisher: Springer
publist_id: '6932'
pubrep_id: '881'
quality_controlled: '1'
scopus_import: '1'
status: public
title: The number and distribution of AMPA receptor channels containing fast kinetic
  GluA3 and GluA4 subunits at auditory nerve synapses depend on the target cells
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 222
year: '2017'
...
---
_id: '1094'
abstract:
- lang: eng
  text: Immunogold labeling of freeze-fracture replicas has recently been used for
    high-resolution visualization of protein localization in electron microscopy.
    This method has higher labeling efficiency than conventional immunogold methods
    for membrane molecules allowing precise quantitative measurements. However, one
    of the limitations of freeze-fracture replica immunolabeling is difficulty in
    keeping structural orientation and identifying labeled profiles in complex tissues
    like brain. The difficulty is partly due to fragmentation of freeze-fracture replica
    preparations during labeling procedures and limited morphological clues on the
    replica surface. To overcome these issues, we introduce here a grid-glued replica
    method combined with SEM observation. This method allows histological staining
    before dissolving the tissue and easy handling of replicas during immunogold labeling,
    and keeps the whole replica surface intact without fragmentation. The procedure
    described here is also useful for matched double-replica analysis allowing further
    identification of labeled profiles in corresponding P-face and E-face.
acknowledged_ssus:
- _id: EM-Fac
acknowledgement: 'We thank Prof. Elek Molnár for providing us a pan-AMPAR anti-body
  used in Fig.2 and Dr. Ludek Lovicar for technical assistance in scanning electron
  microscope imaging. This work was supported by the European Union (HBP—Project Ref.
  604102). '
alternative_title:
- Methods in Molecular Biology
article_processing_charge: No
author:
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
citation:
  ama: 'Harada H, Shigemoto R. Immunogold protein localization on grid-glued freeze-fracture
    replicas. In: <i>High-Resolution Imaging of Cellular Proteins</i>. Vol 1474. Springer;
    2016:203-216. doi:<a href="https://doi.org/10.1007/978-1-4939-6352-2_12">10.1007/978-1-4939-6352-2_12</a>'
  apa: Harada, H., &#38; Shigemoto, R. (2016). Immunogold protein localization on
    grid-glued freeze-fracture replicas. In <i>High-Resolution Imaging of Cellular
    Proteins</i> (Vol. 1474, pp. 203–216). Springer. <a href="https://doi.org/10.1007/978-1-4939-6352-2_12">https://doi.org/10.1007/978-1-4939-6352-2_12</a>
  chicago: Harada, Harumi, and Ryuichi Shigemoto. “Immunogold Protein Localization
    on Grid-Glued Freeze-Fracture Replicas.” In <i>High-Resolution Imaging of Cellular
    Proteins</i>, 1474:203–16. Springer, 2016. <a href="https://doi.org/10.1007/978-1-4939-6352-2_12">https://doi.org/10.1007/978-1-4939-6352-2_12</a>.
  ieee: H. Harada and R. Shigemoto, “Immunogold protein localization on grid-glued
    freeze-fracture replicas,” in <i>High-Resolution Imaging of Cellular Proteins</i>,
    vol. 1474, Springer, 2016, pp. 203–216.
  ista: 'Harada H, Shigemoto R. 2016.Immunogold protein localization on grid-glued
    freeze-fracture replicas. In: High-Resolution Imaging of Cellular Proteins. Methods
    in Molecular Biology, vol. 1474, 203–216.'
  mla: Harada, Harumi, and Ryuichi Shigemoto. “Immunogold Protein Localization on
    Grid-Glued Freeze-Fracture Replicas.” <i>High-Resolution Imaging of Cellular Proteins</i>,
    vol. 1474, Springer, 2016, pp. 203–16, doi:<a href="https://doi.org/10.1007/978-1-4939-6352-2_12">10.1007/978-1-4939-6352-2_12</a>.
  short: H. Harada, R. Shigemoto, in:, High-Resolution Imaging of Cellular Proteins,
    Springer, 2016, pp. 203–216.
date_created: 2018-12-11T11:50:06Z
date_published: 2016-08-12T00:00:00Z
date_updated: 2023-09-05T14:09:01Z
day: '12'
department:
- _id: RySh
doi: 10.1007/978-1-4939-6352-2_12
ec_funded: 1
intvolume: '      1474'
language:
- iso: eng
month: '08'
oa_version: None
page: 203 - 216
project:
- _id: 25CD3DD2-B435-11E9-9278-68D0E5697425
  call_identifier: FP7
  grant_number: '604102'
  name: Localization of ion channels and receptors by two and three-dimensional immunoelectron
    microscopic approaches
publication: High-Resolution Imaging of Cellular Proteins
publication_identifier:
  eissn:
  - 1611-3349
  issn:
  - 0302-9743
publication_status: published
publisher: Springer
publist_id: '6281'
quality_controlled: '1'
status: public
title: Immunogold protein localization on grid-glued freeze-fracture replicas
type: book_chapter
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 1474
year: '2016'
...
---
_id: '1546'
abstract:
- lang: eng
  text: Synaptic efficacy and precision are influenced by the coupling of voltage-gated
    Ca2+ channels (VGCCs) to vesicles. But because the topography of VGCCs and their
    proximity to vesicles is unknown, a quantitative understanding of the determinants
    of vesicular release at nanometer scale is lacking. To investigate this, we combined
    freeze-fracture replica immunogold labeling of Cav2.1 channels, local [Ca2+] imaging,
    and patch pipette perfusion of EGTA at the calyx of Held. Between postnatal day
    7 and 21, VGCCs formed variable sized clusters and vesicular release became less
    sensitive to EGTA, whereas fixed Ca2+ buffer properties remained constant. Experimentally
    constrained reaction-diffusion simulations suggest that Ca2+ sensors for vesicular
    release are located at the perimeter of VGCC clusters (&lt;30nm) and predict that
    VGCC number per cluster determines vesicular release probability without altering
    release time course. This &quot;perimeter release model&quot; provides a unifying
    framework accounting for developmental changes in both synaptic efficacy and time
    course.
acknowledgement: This work was supported by the Core Research for Evolutional Science
  and Technology (CREST) of Japan Science and Technology Agency to T.T. and R.S.;
  by the funding provided by Okinawa Institute of Science and Technology (OIST) to
  T.T. and Y.N.; by JSPS Core-to-Core Program, A. Advanced Networks to T.T.; by the
  Grant-in-Aid for Young Scientists from the Japanese Ministry of Education, Culture,
  Sports, Science and Technology (#23700474) to Y.N.; by the Centre National de la
  Recherche Scientifique through the Actions Thematiques et Initatives sur Programme,
  Fondation Fyssen, Fondation pour la Recherche Medicale, Federation pour la Recherche
  sur le Cerveau, Agence Nationale de la Recherche (ANR-2007-Neuro-008-01 and ANR-2010-BLAN-1411-01)
  to D.D. and Y.N.; and by the European Commission Coordination Action ENINET (LSHM-CT-2005-19063)
  to D.D. and R.A.S. R.A.S. and J.S.R. were funded by Wellcome Trust Senior (064413)
  and Principal (095667) Research Fellowship and an ERC advance grant (294667) to
  RAS.
author:
- first_name: Yukihiro
  full_name: Nakamura, Yukihiro
  last_name: Nakamura
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Naomi
  full_name: Kamasawa, Naomi
  last_name: Kamasawa
- first_name: Ko
  full_name: Matsui, Ko
  last_name: Matsui
- first_name: Jason
  full_name: Rothman, Jason
  last_name: Rothman
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: R Angus
  full_name: Silver, R Angus
  last_name: Silver
- first_name: David
  full_name: Digregorio, David
  last_name: Digregorio
- first_name: Tomoyuki
  full_name: Takahashi, Tomoyuki
  last_name: Takahashi
citation:
  ama: Nakamura Y, Harada H, Kamasawa N, et al. Nanoscale distribution of presynaptic
    Ca2+ channels and its impact on vesicular release during development. <i>Neuron</i>.
    2015;85(1):145-158. doi:<a href="https://doi.org/10.1016/j.neuron.2014.11.019">10.1016/j.neuron.2014.11.019</a>
  apa: Nakamura, Y., Harada, H., Kamasawa, N., Matsui, K., Rothman, J., Shigemoto,
    R., … Takahashi, T. (2015). Nanoscale distribution of presynaptic Ca2+ channels
    and its impact on vesicular release during development. <i>Neuron</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.neuron.2014.11.019">https://doi.org/10.1016/j.neuron.2014.11.019</a>
  chicago: Nakamura, Yukihiro, Harumi Harada, Naomi Kamasawa, Ko Matsui, Jason Rothman,
    Ryuichi Shigemoto, R Angus Silver, David Digregorio, and Tomoyuki Takahashi. “Nanoscale
    Distribution of Presynaptic Ca2+ Channels and Its Impact on Vesicular Release
    during Development.” <i>Neuron</i>. Elsevier, 2015. <a href="https://doi.org/10.1016/j.neuron.2014.11.019">https://doi.org/10.1016/j.neuron.2014.11.019</a>.
  ieee: Y. Nakamura <i>et al.</i>, “Nanoscale distribution of presynaptic Ca2+ channels
    and its impact on vesicular release during development,” <i>Neuron</i>, vol. 85,
    no. 1. Elsevier, pp. 145–158, 2015.
  ista: Nakamura Y, Harada H, Kamasawa N, Matsui K, Rothman J, Shigemoto R, Silver
    RA, Digregorio D, Takahashi T. 2015. Nanoscale distribution of presynaptic Ca2+
    channels and its impact on vesicular release during development. Neuron. 85(1),
    145–158.
  mla: Nakamura, Yukihiro, et al. “Nanoscale Distribution of Presynaptic Ca2+ Channels
    and Its Impact on Vesicular Release during Development.” <i>Neuron</i>, vol. 85,
    no. 1, Elsevier, 2015, pp. 145–58, doi:<a href="https://doi.org/10.1016/j.neuron.2014.11.019">10.1016/j.neuron.2014.11.019</a>.
  short: Y. Nakamura, H. Harada, N. Kamasawa, K. Matsui, J. Rothman, R. Shigemoto,
    R.A. Silver, D. Digregorio, T. Takahashi, Neuron 85 (2015) 145–158.
date_created: 2018-12-11T11:52:39Z
date_published: 2015-01-07T00:00:00Z
date_updated: 2021-01-12T06:51:31Z
day: '07'
ddc:
- '570'
department:
- _id: RySh
doi: 10.1016/j.neuron.2014.11.019
file:
- access_level: open_access
  checksum: 725f4d5be2dbb44b283ce722645ef37d
  content_type: application/pdf
  creator: system
  date_created: 2018-12-12T10:15:47Z
  date_updated: 2020-07-14T12:45:01Z
  file_id: '5170'
  file_name: IST-2016-482-v1+1_1-s2.0-S0896627314010472-main.pdf
  file_size: 3080111
  relation: main_file
file_date_updated: 2020-07-14T12:45:01Z
has_accepted_license: '1'
intvolume: '        85'
issue: '1'
language:
- iso: eng
month: '01'
oa: 1
oa_version: Published Version
page: 145 - 158
publication: Neuron
publication_status: published
publisher: Elsevier
publist_id: '5625'
pubrep_id: '482'
quality_controlled: '1'
scopus_import: 1
status: public
title: Nanoscale distribution of presynaptic Ca2+ channels and its impact on vesicular
  release during development
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 85
year: '2015'
...
---
_id: '1898'
abstract:
- lang: eng
  text: Fast synaptic transmission is important for rapid information processing.
    To explore the maximal rate of neuronal signaling and to analyze the presynaptic
    mechanisms, we focused on the input layer of the cerebellar cortex, where exceptionally
    high action potential (AP) frequencies have been reported invivo. With paired
    recordings between presynaptic cerebellar mossy fiber boutons and postsynaptic
    granule cells, we demonstrate reliable neurotransmission upto ~1 kHz. Presynaptic
    APs are ultrafast, with ~100μs half-duration. Both Kv1 and Kv3 potassium channels
    mediate the fast repolarization, rapidly inactivating sodium channels ensure metabolic
    efficiency, and little AP broadening occurs during bursts of up to 1.5 kHz. Presynaptic
    Cav2.1 (P/Q-type) calcium channels open efficiently during ultrafast APs. Furthermore,
    a subset of synaptic vesicles is tightly coupled to Ca2+ channels, and vesicles
    are rapidly recruited to the release site. These data reveal mechanisms of presynaptic
    AP generation and transmitter release underlying neuronal kHz signaling.
author:
- first_name: Andreas
  full_name: Ritzau Jost, Andreas
  last_name: Ritzau Jost
- first_name: Igor
  full_name: Delvendahl, Igor
  last_name: Delvendahl
- first_name: Annika
  full_name: Rings, Annika
  last_name: Rings
- first_name: Niklas
  full_name: Byczkowicz, Niklas
  last_name: Byczkowicz
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Johannes
  full_name: Hirrlinger, Johannes
  last_name: Hirrlinger
- first_name: Jens
  full_name: Eilers, Jens
  last_name: Eilers
- first_name: Stefan
  full_name: Hallermann, Stefan
  last_name: Hallermann
citation:
  ama: Ritzau Jost A, Delvendahl I, Rings A, et al. Ultrafast action potentials mediate
    kilohertz signaling at a central synapse. <i>Neuron</i>. 2014;84(1):152-163. doi:<a
    href="https://doi.org/10.1016/j.neuron.2014.08.036">10.1016/j.neuron.2014.08.036</a>
  apa: Ritzau Jost, A., Delvendahl, I., Rings, A., Byczkowicz, N., Harada, H., Shigemoto,
    R., … Hallermann, S. (2014). Ultrafast action potentials mediate kilohertz signaling
    at a central synapse. <i>Neuron</i>. Elsevier. <a href="https://doi.org/10.1016/j.neuron.2014.08.036">https://doi.org/10.1016/j.neuron.2014.08.036</a>
  chicago: Ritzau Jost, Andreas, Igor Delvendahl, Annika Rings, Niklas Byczkowicz,
    Harumi Harada, Ryuichi Shigemoto, Johannes Hirrlinger, Jens Eilers, and Stefan
    Hallermann. “Ultrafast Action Potentials Mediate Kilohertz Signaling at a Central
    Synapse.” <i>Neuron</i>. Elsevier, 2014. <a href="https://doi.org/10.1016/j.neuron.2014.08.036">https://doi.org/10.1016/j.neuron.2014.08.036</a>.
  ieee: A. Ritzau Jost <i>et al.</i>, “Ultrafast action potentials mediate kilohertz
    signaling at a central synapse,” <i>Neuron</i>, vol. 84, no. 1. Elsevier, pp.
    152–163, 2014.
  ista: Ritzau Jost A, Delvendahl I, Rings A, Byczkowicz N, Harada H, Shigemoto R,
    Hirrlinger J, Eilers J, Hallermann S. 2014. Ultrafast action potentials mediate
    kilohertz signaling at a central synapse. Neuron. 84(1), 152–163.
  mla: Ritzau Jost, Andreas, et al. “Ultrafast Action Potentials Mediate Kilohertz
    Signaling at a Central Synapse.” <i>Neuron</i>, vol. 84, no. 1, Elsevier, 2014,
    pp. 152–63, doi:<a href="https://doi.org/10.1016/j.neuron.2014.08.036">10.1016/j.neuron.2014.08.036</a>.
  short: A. Ritzau Jost, I. Delvendahl, A. Rings, N. Byczkowicz, H. Harada, R. Shigemoto,
    J. Hirrlinger, J. Eilers, S. Hallermann, Neuron 84 (2014) 152–163.
date_created: 2018-12-11T11:54:36Z
date_published: 2014-10-01T00:00:00Z
date_updated: 2021-01-12T06:53:55Z
day: '01'
department:
- _id: RySh
doi: 10.1016/j.neuron.2014.08.036
intvolume: '        84'
issue: '1'
language:
- iso: eng
month: '10'
oa_version: None
page: 152 - 163
publication: Neuron
publication_status: published
publisher: Elsevier
publist_id: '5197'
quality_controlled: '1'
scopus_import: 1
status: public
title: Ultrafast action potentials mediate kilohertz signaling at a central synapse
type: journal_article
user_id: 4435EBFC-F248-11E8-B48F-1D18A9856A87
volume: 84
year: '2014'
...
---
_id: '2478'
abstract:
- lang: eng
  text: Despite the pivotal functions of the NMDA receptor (NMDAR) for neural circuit
    development and synaptic plasticity, the molecular mechanisms underlying the dynamics
    of NMDAR trafficking are poorly understood. The cell adhesion molecule neuroligin-1
    (NL1) modifies NMDAR-dependent synaptic transmission and synaptic plasticity,
    but it is unclear whether NL1 controls synaptic accumulation or function of the
    receptors. Here, we provide evidence that NL1 regulates the abundance of NMDARs
    at postsynaptic sites. This function relies on extracellular, NL1 isoform-specific
    sequences that facilitate biochemical interactions between NL1 and the NMDAR GluN1
    subunit. Our work uncovers NL1 isoform-specific cisinteractions with ionotropic
    glutamate receptors as a key mechanism for controlling synaptic properties.
author:
- first_name: Elaine
  full_name: Budreck, Elaine C
  last_name: Budreck
- first_name: Oh
  full_name: Kwon, Oh-Bin
  last_name: Kwon
- first_name: Jung
  full_name: Jung, Jung-Hoon
  last_name: Jung
- first_name: Stéphane
  full_name: Baudouin, Stéphane J
  last_name: Baudouin
- first_name: Albert
  full_name: Thommen, Albert
  last_name: Thommen
- first_name: Hye
  full_name: Kim, Hye-Sun
  last_name: Kim
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
- first_name: Harumi
  full_name: Harumi Harada
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Katsuhiko
  full_name: Tabuchi, Katsuhiko
  last_name: Tabuchi
- first_name: Ryuichi
  full_name: Ryuichi Shigemoto
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Peter
  full_name: Scheiffele, Peter
  last_name: Scheiffele
- first_name: Joung
  full_name: Kim, Joung-Hun
  last_name: Kim
citation:
  ama: Budreck E, Kwon O, Jung J, et al. Neuroligin-1 controls synaptic abundance
    of NMDA-type glutamate receptors through extracellular coupling. <i>PNAS</i>.
    2013;110(2):725-730. doi:<a href="https://doi.org/10.1073/pnas.1214718110">10.1073/pnas.1214718110</a>
  apa: Budreck, E., Kwon, O., Jung, J., Baudouin, S., Thommen, A., Kim, H., … Kim,
    J. (2013). Neuroligin-1 controls synaptic abundance of NMDA-type glutamate receptors
    through extracellular coupling. <i>PNAS</i>. National Academy of Sciences. <a
    href="https://doi.org/10.1073/pnas.1214718110">https://doi.org/10.1073/pnas.1214718110</a>
  chicago: Budreck, Elaine, Oh Kwon, Jung Jung, Stéphane Baudouin, Albert Thommen,
    Hye Kim, Yugo Fukazawa, et al. “Neuroligin-1 Controls Synaptic Abundance of NMDA-Type
    Glutamate Receptors through Extracellular Coupling.” <i>PNAS</i>. National Academy
    of Sciences, 2013. <a href="https://doi.org/10.1073/pnas.1214718110">https://doi.org/10.1073/pnas.1214718110</a>.
  ieee: E. Budreck <i>et al.</i>, “Neuroligin-1 controls synaptic abundance of NMDA-type
    glutamate receptors through extracellular coupling,” <i>PNAS</i>, vol. 110, no.
    2. National Academy of Sciences, pp. 725–730, 2013.
  ista: Budreck E, Kwon O, Jung J, Baudouin S, Thommen A, Kim H, Fukazawa Y, Harada
    H, Tabuchi K, Shigemoto R, Scheiffele P, Kim J. 2013. Neuroligin-1 controls synaptic
    abundance of NMDA-type glutamate receptors through extracellular coupling. PNAS.
    110(2), 725–730.
  mla: Budreck, Elaine, et al. “Neuroligin-1 Controls Synaptic Abundance of NMDA-Type
    Glutamate Receptors through Extracellular Coupling.” <i>PNAS</i>, vol. 110, no.
    2, National Academy of Sciences, 2013, pp. 725–30, doi:<a href="https://doi.org/10.1073/pnas.1214718110">10.1073/pnas.1214718110</a>.
  short: E. Budreck, O. Kwon, J. Jung, S. Baudouin, A. Thommen, H. Kim, Y. Fukazawa,
    H. Harada, K. Tabuchi, R. Shigemoto, P. Scheiffele, J. Kim, PNAS 110 (2013) 725–730.
date_created: 2018-12-11T11:57:54Z
date_published: 2013-01-08T00:00:00Z
date_updated: 2021-01-12T06:57:43Z
day: '08'
doi: 10.1073/pnas.1214718110
extern: 1
intvolume: '       110'
issue: '2'
month: '01'
page: 725 - 730
publication: PNAS
publication_status: published
publisher: National Academy of Sciences
publist_id: '4423'
quality_controlled: 0
status: public
title: Neuroligin-1 controls synaptic abundance of NMDA-type glutamate receptors through
  extracellular coupling
type: journal_article
volume: 110
year: '2013'
...
---
_id: '2690'
abstract:
- lang: eng
  text: Establishing the spatiotemporal concentration profile of neurotransmitter
    following synaptic vesicular release is essential for our understanding of inter-neuronal
    communication. Such profile is a determinant of synaptic strength, short-term
    plasticity and inter-synaptic crosstalk. Synaptically released glutamate has been
    suggested to reach a few millimolar in concentration and last for &lt;1 ms. The
    synaptic cleft is often conceived as a single concentration compartment, whereas
    a huge gradient likely exists. Modelling studies have attempted to describe this
    gradient, but two key parameters, the number of glutamate in a vesicle (NGlu)
    and its diffusion coefficient (DGlu) in the extracellular space, remained unresolved.
    To determine this profile, the rat calyx of Held synapse at postnatal day 12-16
    was studied where diffusion of glutamate occurs two-dimensionally and where quantification
    of AMPA receptor distribution on individual postsynaptic specialization on medial
    nucleus of the trapezoid body principal cells is possible using SDS-digested freeze-fracture
    replica labelling. To assess the performance of these receptors as glutamate sensors,
    a kinetic model of the receptors was constructed from outside-out patch recordings.
    From here, we simulated synaptic responses and compared them with the EPSC recordings.
    Combinations of NGlu and DGlu with an optimum of 7000 and 0.3 μm2 ms-1 reproduced
    the data, suggesting slow diffusion. Further simulations showed that a single
    vesicle does not saturate the synaptic receptors, and that glutamate spillover
    does not affect the conductance amplitude at this synapse. Using the estimated
    profile, we also evaluated how the number of multiple vesicle releases at individual
    active zones affects the amplitude of postsynaptic signals.
author:
- first_name: Timotheus
  full_name: Budisantoso, Timotheus
  last_name: Budisantoso
- first_name: Harumi
  full_name: Harumi Harada
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Naomi
  full_name: Kamasawa, Naomi
  last_name: Kamasawa
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
- first_name: Ryuichi
  full_name: Ryuichi Shigemoto
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Ko
  full_name: Matsui, Ko
  last_name: Matsui
citation:
  ama: Budisantoso T, Harada H, Kamasawa N, Fukazawa Y, Shigemoto R, Matsui K. Evaluation
    of glutamate concentration transient in the synaptic cleft of the rat calyx of
    Held. <i>Journal of Physiology</i>. 2013;591(1):219-239. doi:<a href="https://doi.org/10.1113/jphysiol.2012.241398">10.1113/jphysiol.2012.241398</a>
  apa: Budisantoso, T., Harada, H., Kamasawa, N., Fukazawa, Y., Shigemoto, R., &#38;
    Matsui, K. (2013). Evaluation of glutamate concentration transient in the synaptic
    cleft of the rat calyx of Held. <i>Journal of Physiology</i>. Wiley-Blackwell.
    <a href="https://doi.org/10.1113/jphysiol.2012.241398">https://doi.org/10.1113/jphysiol.2012.241398</a>
  chicago: Budisantoso, Timotheus, Harumi Harada, Naomi Kamasawa, Yugo Fukazawa, Ryuichi
    Shigemoto, and Ko Matsui. “Evaluation of Glutamate Concentration Transient in
    the Synaptic Cleft of the Rat Calyx of Held.” <i>Journal of Physiology</i>. Wiley-Blackwell,
    2013. <a href="https://doi.org/10.1113/jphysiol.2012.241398">https://doi.org/10.1113/jphysiol.2012.241398</a>.
  ieee: T. Budisantoso, H. Harada, N. Kamasawa, Y. Fukazawa, R. Shigemoto, and K.
    Matsui, “Evaluation of glutamate concentration transient in the synaptic cleft
    of the rat calyx of Held,” <i>Journal of Physiology</i>, vol. 591, no. 1. Wiley-Blackwell,
    pp. 219–239, 2013.
  ista: Budisantoso T, Harada H, Kamasawa N, Fukazawa Y, Shigemoto R, Matsui K. 2013.
    Evaluation of glutamate concentration transient in the synaptic cleft of the rat
    calyx of Held. Journal of Physiology. 591(1), 219–239.
  mla: Budisantoso, Timotheus, et al. “Evaluation of Glutamate Concentration Transient
    in the Synaptic Cleft of the Rat Calyx of Held.” <i>Journal of Physiology</i>,
    vol. 591, no. 1, Wiley-Blackwell, 2013, pp. 219–39, doi:<a href="https://doi.org/10.1113/jphysiol.2012.241398">10.1113/jphysiol.2012.241398</a>.
  short: T. Budisantoso, H. Harada, N. Kamasawa, Y. Fukazawa, R. Shigemoto, K. Matsui,
    Journal of Physiology 591 (2013) 219–239.
date_created: 2018-12-11T11:59:05Z
date_published: 2013-01-01T00:00:00Z
date_updated: 2021-01-12T06:59:04Z
day: '01'
doi: 10.1113/jphysiol.2012.241398
extern: 1
intvolume: '       591'
issue: '1'
month: '01'
page: 219 - 239
publication: Journal of Physiology
publication_status: published
publisher: Wiley-Blackwell
publist_id: '4207'
quality_controlled: 0
status: public
title: Evaluation of glutamate concentration transient in the synaptic cleft of the
  rat calyx of Held
type: journal_article
volume: 591
year: '2013'
...
---
_id: '2615'
abstract:
- lang: eng
  text: Taste-mGluR4, cloned from taste tissues, is a truncated variant of brain-expressed
    mGluR4a (brain-mGluR4), and is known to be a candidate for the receptor involved
    in the umami taste sense. Although the expression patterns of taste- and brain-mGluR4
    mRNAs have been demonstrated, no mention has so far been made of the expression
    of these two mGluR4 proteins in taste tissues. The present study examined the
    expression of taste-mGluR4 and brain-mGluR4 proteins in rat taste tissues by using
    a specific antibody for mGluR4a which shared a C-terminus of both taste- and brain-mGluR4,
    for immunoblot analysis and immunohistochemistry. Immunoblot analysis showed that
    both brain-mGluR4 and taste-mGluR4 were expressed in the taste tissues. Taste-mGluR4
    was not detected in the cerebellum. The immunoreactive band for brain-mGluR4 protein
    was much stronger than that for taste-mGluR4 protein. In the cryosections of fungiform,
    foliate and circumvallate papillae, the antibody against taste-mGluR4 exhibited
    intense labeling of the taste pores and taste hairs in all the taste buds of gustatory
    papillae examined; the immunoreaction to the antibody against brain-mGluR4 was
    more intense at the same sites of the taste buds. The portions of the taste bud
    cells below the taste pore and surrounding keratinocytes did not show any immunoreactivities.
    The results of the present study strongly suggest that, in addition to taste-mGluR4,
    brain-mGluR4 may function even more importantly than the former as a receptor
    for glutamate, i.e. the umami taste sensation.
article_processing_charge: No
article_type: original
author:
- first_name: Takashi
  full_name: Toyono, Takashi
  last_name: Toyono
- first_name: Yuji
  full_name: Seta, Yuji
  last_name: Seta
- first_name: Shinji
  full_name: Sataoka, Shinji
  last_name: Sataoka
- first_name: Harumi
  full_name: Harada, Harumi
  id: 2E55CDF2-F248-11E8-B48F-1D18A9856A87
  last_name: Harada
  orcid: 0000-0001-7429-7896
- first_name: Takahiko
  full_name: Morotomi, Takahiko
  last_name: Morotomi
- first_name: Shintaro
  full_name: Kawano, Shintaro
  last_name: Kawano
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Kuniaki
  full_name: Toyoshima, Kuniaki
  last_name: Toyoshima
citation:
  ama: Toyono T, Seta Y, Sataoka S, et al. Expression of the metabotropic glutamate
    receptor, mGluR4a, in the taste hairs of taste buds in rat gustatory papillae.
    <i>Archives of Histology and Cytology</i>. 2002;65(1):91-96. doi:<a href="https://doi.org/10.1679/aohc.65.91">10.1679/aohc.65.91</a>
  apa: Toyono, T., Seta, Y., Sataoka, S., Harada, H., Morotomi, T., Kawano, S., …
    Toyoshima, K. (2002). Expression of the metabotropic glutamate receptor, mGluR4a,
    in the taste hairs of taste buds in rat gustatory papillae. <i>Archives of Histology
    and Cytology</i>. Japan Society of Histological Documentation. <a href="https://doi.org/10.1679/aohc.65.91">https://doi.org/10.1679/aohc.65.91</a>
  chicago: Toyono, Takashi, Yuji Seta, Shinji Sataoka, Harumi Harada, Takahiko Morotomi,
    Shintaro Kawano, Ryuichi Shigemoto, and Kuniaki Toyoshima. “Expression of the
    Metabotropic Glutamate Receptor, MGluR4a, in the Taste Hairs of Taste Buds in
    Rat Gustatory Papillae.” <i>Archives of Histology and Cytology</i>. Japan Society
    of Histological Documentation, 2002. <a href="https://doi.org/10.1679/aohc.65.91">https://doi.org/10.1679/aohc.65.91</a>.
  ieee: T. Toyono <i>et al.</i>, “Expression of the metabotropic glutamate receptor,
    mGluR4a, in the taste hairs of taste buds in rat gustatory papillae,” <i>Archives
    of Histology and Cytology</i>, vol. 65, no. 1. Japan Society of Histological Documentation,
    pp. 91–96, 2002.
  ista: Toyono T, Seta Y, Sataoka S, Harada H, Morotomi T, Kawano S, Shigemoto R,
    Toyoshima K. 2002. Expression of the metabotropic glutamate receptor, mGluR4a,
    in the taste hairs of taste buds in rat gustatory papillae. Archives of Histology
    and Cytology. 65(1), 91–96.
  mla: Toyono, Takashi, et al. “Expression of the Metabotropic Glutamate Receptor,
    MGluR4a, in the Taste Hairs of Taste Buds in Rat Gustatory Papillae.” <i>Archives
    of Histology and Cytology</i>, vol. 65, no. 1, Japan Society of Histological Documentation,
    2002, pp. 91–96, doi:<a href="https://doi.org/10.1679/aohc.65.91">10.1679/aohc.65.91</a>.
  short: T. Toyono, Y. Seta, S. Sataoka, H. Harada, T. Morotomi, S. Kawano, R. Shigemoto,
    K. Toyoshima, Archives of Histology and Cytology 65 (2002) 91–96.
date_created: 2018-12-11T11:58:41Z
date_published: 2002-01-01T00:00:00Z
date_updated: 2023-07-25T10:00:15Z
day: '01'
doi: 10.1679/aohc.65.91
extern: '1'
external_id:
  pmid:
  - '12002614'
intvolume: '        65'
issue: '1'
language:
- iso: eng
month: '01'
oa_version: None
page: 91 - 96
pmid: 1
publication: Archives of Histology and Cytology
publication_identifier:
  issn:
  - 0914-9465
publication_status: published
publisher: Japan Society of Histological Documentation
publist_id: '4283'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Expression of the metabotropic glutamate receptor, mGluR4a, in the taste hairs
  of taste buds in rat gustatory papillae
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 65
year: '2002'
...