---
_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: '7207'
abstract:
- lang: eng
  text: The hippocampus plays key roles in learning and memory and is a main target
    of Alzheimer's disease (AD), which causes progressive memory impairments. Despite
    numerous investigations about the processes required for the normal hippocampal
    functions, the neurotransmitter receptors involved in the synaptic deficits by
    which AD disables the hippocampus are not yet characterized. By combining histoblots,
    western blots, immunohistochemistry and high‐resolution immunoelectron microscopic
    methods for GABAB receptors, this study provides a quantitative description of
    the expression and the subcellular localization of GABAB1 in the hippocampus in
    a mouse model of AD at 1, 6 and 12 months of age. Western blots and histoblots
    showed that the total amount of protein and the laminar expression pattern of
    GABAB1 were similar in APP/PS1 mice and in age‐matched wild‐type mice. In contrast,
    immunoelectron microscopic techniques showed that the subcellular localization
    of GABAB1 subunit did not change significantly in APP/PS1 mice at 1 month of age,
    was significantly reduced in the stratum lacunosum‐moleculare of CA1 pyramidal
    cells at 6 months of age and significantly reduced at the membrane surface of
    CA1 pyramidal cells at 12 months of age. This reduction of plasma membrane GABAB1
    was paralleled by a significant increase of the subunit at the intracellular sites.
    We further observed a decrease of membrane‐targeted GABAB receptors in axon terminals
    contacting CA1 pyramidal cells. Our data demonstrate compartment‐ and age‐dependent
    reduction of plasma membrane‐targeted GABAB receptors in the CA1 region of the
    hippocampus, suggesting that this decrease might be enough to alter the GABAB‐mediated
    synaptic transmission taking place in AD.
article_processing_charge: No
article_type: original
author:
- first_name: Alejandro
  full_name: Martín-Belmonte, Alejandro
  last_name: Martín-Belmonte
- first_name: Carolina
  full_name: Aguado, Carolina
  last_name: Aguado
- first_name: Rocío
  full_name: Alfaro-Ruíz, Rocío
  last_name: Alfaro-Ruíz
- first_name: Ana Esther
  full_name: Moreno-Martínez, Ana Esther
  last_name: Moreno-Martínez
- first_name: Luis
  full_name: De La Ossa, Luis
  last_name: De La Ossa
- first_name: José
  full_name: Martínez-Hernández, José
  last_name: Martínez-Hernández
- first_name: Alain
  full_name: Buisson, Alain
  last_name: Buisson
- first_name: Simon
  full_name: Früh, Simon
  last_name: Früh
- first_name: Bernhard
  full_name: Bettler, Bernhard
  last_name: Bettler
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
- first_name: Rafael
  full_name: Luján, Rafael
  last_name: Luján
citation:
  ama: Martín-Belmonte A, Aguado C, Alfaro-Ruíz R, et al. Reduction in the neuronal
    surface of post and presynaptic GABA&#62;B&#60; receptors in the hippocampus in
    a mouse model of Alzheimer’s disease. <i>Brain Pathology</i>. 2020;30(3):554-575.
    doi:<a href="https://doi.org/10.1111/bpa.12802">10.1111/bpa.12802</a>
  apa: Martín-Belmonte, A., Aguado, C., Alfaro-Ruíz, R., Moreno-Martínez, A. E., De
    La Ossa, L., Martínez-Hernández, J., … Luján, R. (2020). Reduction in the neuronal
    surface of post and presynaptic GABA&#62;B&#60; receptors in the hippocampus in
    a mouse model of Alzheimer’s disease. <i>Brain Pathology</i>. Wiley. <a href="https://doi.org/10.1111/bpa.12802">https://doi.org/10.1111/bpa.12802</a>
  chicago: Martín-Belmonte, Alejandro, Carolina Aguado, Rocío Alfaro-Ruíz, Ana Esther
    Moreno-Martínez, Luis De La Ossa, José Martínez-Hernández, Alain Buisson, et al.
    “Reduction in the Neuronal Surface of Post and Presynaptic GABA&#62;B&#60; Receptors
    in the Hippocampus in a Mouse Model of Alzheimer’s Disease.” <i>Brain Pathology</i>.
    Wiley, 2020. <a href="https://doi.org/10.1111/bpa.12802">https://doi.org/10.1111/bpa.12802</a>.
  ieee: A. Martín-Belmonte <i>et al.</i>, “Reduction in the neuronal surface of post
    and presynaptic GABA&#62;B&#60; receptors in the hippocampus in a mouse model
    of Alzheimer’s disease,” <i>Brain Pathology</i>, vol. 30, no. 3. Wiley, pp. 554–575,
    2020.
  ista: Martín-Belmonte A, Aguado C, Alfaro-Ruíz R, Moreno-Martínez AE, De La Ossa
    L, Martínez-Hernández J, Buisson A, Früh S, Bettler B, Shigemoto R, Fukazawa Y,
    Luján R. 2020. Reduction in the neuronal surface of post and presynaptic GABA&#62;B&#60;
    receptors in the hippocampus in a mouse model of Alzheimer’s disease. Brain Pathology.
    30(3), 554–575.
  mla: Martín-Belmonte, Alejandro, et al. “Reduction in the Neuronal Surface of Post
    and Presynaptic GABA&#62;B&#60; Receptors in the Hippocampus in a Mouse Model
    of Alzheimer’s Disease.” <i>Brain Pathology</i>, vol. 30, no. 3, Wiley, 2020,
    pp. 554–75, doi:<a href="https://doi.org/10.1111/bpa.12802">10.1111/bpa.12802</a>.
  short: A. Martín-Belmonte, C. Aguado, R. Alfaro-Ruíz, A.E. Moreno-Martínez, L. De
    La Ossa, J. Martínez-Hernández, A. Buisson, S. Früh, B. Bettler, R. Shigemoto,
    Y. Fukazawa, R. Luján, Brain Pathology 30 (2020) 554–575.
date_created: 2019-12-22T23:00:43Z
date_published: 2020-05-01T00:00:00Z
date_updated: 2023-09-06T14:48:01Z
day: '01'
ddc:
- '570'
department:
- _id: RySh
doi: 10.1111/bpa.12802
ec_funded: 1
external_id:
  isi:
  - '000502270900001'
  pmid:
  - '31729777'
file:
- access_level: open_access
  checksum: 549cc1b18f638a21d17a939ba5563fa9
  content_type: application/pdf
  creator: dernst
  date_created: 2020-09-22T09:47:19Z
  date_updated: 2020-09-22T09:47:19Z
  file_id: '8554'
  file_name: 2020_BrainPathology_MartinBelmonte.pdf
  file_size: 4220935
  relation: main_file
  success: 1
file_date_updated: 2020-09-22T09:47:19Z
has_accepted_license: '1'
intvolume: '        30'
isi: 1
issue: '3'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
page: 554-575
pmid: 1
project:
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
- _id: 26436750-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '785907'
  name: Human Brain Project Specific Grant Agreement 2 (HBP SGA 2)
publication: Brain Pathology
publication_identifier:
  eissn:
  - '17503639'
  issn:
  - '10156305'
publication_status: published
publisher: Wiley
quality_controlled: '1'
scopus_import: '1'
status: public
title: Reduction in the neuronal surface of post and presynaptic GABA>B< receptors
  in the hippocampus in a mouse model of Alzheimer's disease
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: 30
year: '2020'
...
---
_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: '292'
abstract:
- lang: eng
  text: 'Retina is a paradigmatic system for studying sensory encoding: the transformation
    of light into spiking activity of ganglion cells. The inverse problem, where stimulus
    is reconstructed from spikes, has received less attention, especially for complex
    stimuli that should be reconstructed “pixel-by-pixel”. We recorded around a hundred
    neurons from a dense patch in a rat retina and decoded movies of multiple small
    randomly-moving discs. We constructed nonlinear (kernelized and neural network)
    decoders that improved significantly over linear results. An important contribution
    to this was the ability of nonlinear decoders to reliably separate between neural
    responses driven by locally fluctuating light signals, and responses at locally
    constant light driven by spontaneous-like activity. This improvement crucially
    depended on the precise, non-Poisson temporal structure of individual spike trains,
    which originated in the spike-history dependence of neural responses. We propose
    a general principle by which downstream circuitry could discriminate between spontaneous
    and stimulus-driven activity based solely on higher-order statistical structure
    in the incoming spike trains.'
article_number: e1006057
article_processing_charge: Yes
article_type: original
author:
- first_name: Vicent
  full_name: Botella Soler, Vicent
  id: 421234E8-F248-11E8-B48F-1D18A9856A87
  last_name: Botella Soler
  orcid: 0000-0002-8790-1914
- first_name: Stephane
  full_name: Deny, Stephane
  last_name: Deny
- first_name: Georg S
  full_name: Martius, Georg S
  last_name: Martius
- first_name: Olivier
  full_name: Marre, Olivier
  last_name: Marre
- first_name: Gasper
  full_name: Tkacik, Gasper
  id: 3D494DCA-F248-11E8-B48F-1D18A9856A87
  last_name: Tkacik
  orcid: 0000-0002-6699-1455
citation:
  ama: Botella Soler V, Deny S, Martius GS, Marre O, Tkačik G. Nonlinear decoding
    of a complex movie from the mammalian retina. <i>PLoS Computational Biology</i>.
    2018;14(5). doi:<a href="https://doi.org/10.1371/journal.pcbi.1006057">10.1371/journal.pcbi.1006057</a>
  apa: Botella Soler, V., Deny, S., Martius, G. S., Marre, O., &#38; Tkačik, G. (2018).
    Nonlinear decoding of a complex movie from the mammalian retina. <i>PLoS Computational
    Biology</i>. Public Library of Science. <a href="https://doi.org/10.1371/journal.pcbi.1006057">https://doi.org/10.1371/journal.pcbi.1006057</a>
  chicago: Botella Soler, Vicente, Stephane Deny, Georg S Martius, Olivier Marre,
    and Gašper Tkačik. “Nonlinear Decoding of a Complex Movie from the Mammalian Retina.”
    <i>PLoS Computational Biology</i>. Public Library of Science, 2018. <a href="https://doi.org/10.1371/journal.pcbi.1006057">https://doi.org/10.1371/journal.pcbi.1006057</a>.
  ieee: V. Botella Soler, S. Deny, G. S. Martius, O. Marre, and G. Tkačik, “Nonlinear
    decoding of a complex movie from the mammalian retina,” <i>PLoS Computational
    Biology</i>, vol. 14, no. 5. Public Library of Science, 2018.
  ista: Botella Soler V, Deny S, Martius GS, Marre O, Tkačik G. 2018. Nonlinear decoding
    of a complex movie from the mammalian retina. PLoS Computational Biology. 14(5),
    e1006057.
  mla: Botella Soler, Vicente, et al. “Nonlinear Decoding of a Complex Movie from
    the Mammalian Retina.” <i>PLoS Computational Biology</i>, vol. 14, no. 5, e1006057,
    Public Library of Science, 2018, doi:<a href="https://doi.org/10.1371/journal.pcbi.1006057">10.1371/journal.pcbi.1006057</a>.
  short: V. Botella Soler, S. Deny, G.S. Martius, O. Marre, G. Tkačik, PLoS Computational
    Biology 14 (2018).
date_created: 2018-12-11T11:45:39Z
date_published: 2018-05-10T00:00:00Z
date_updated: 2024-02-21T13:45:25Z
day: '10'
ddc:
- '570'
department:
- _id: GaTk
doi: 10.1371/journal.pcbi.1006057
ec_funded: 1
external_id:
  isi:
  - '000434012100002'
file:
- access_level: open_access
  checksum: 3026f94d235219e15514505fdbadf34e
  content_type: application/pdf
  creator: dernst
  date_created: 2019-02-13T11:07:15Z
  date_updated: 2020-07-14T12:45:53Z
  file_id: '5974'
  file_name: 2018_Plos_Botella_Soler.pdf
  file_size: 3460786
  relation: main_file
file_date_updated: 2020-07-14T12:45:53Z
has_accepted_license: '1'
intvolume: '        14'
isi: 1
issue: '5'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
project:
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
- _id: 254D1A94-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: P 25651-N26
  name: Sensitivity to higher-order statistics in natural scenes
publication: PLoS Computational Biology
publication_status: published
publisher: Public Library of Science
quality_controlled: '1'
related_material:
  link:
  - description: News on IST Homepage
    relation: press_release
    url: https://ist.ac.at/en/news/video-of-moving-discs-reconstructed-from-rat-retinal-neuron-signals/
  record:
  - id: '5584'
    relation: research_data
    status: public
scopus_import: '1'
status: public
title: Nonlinear decoding of a complex movie from the mammalian retina
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: 14
year: '2018'
...
---
_id: '612'
abstract:
- lang: eng
  text: Metabotropic GABAB receptors mediate slow inhibitory effects presynaptically
    and postsynaptically through the modulation of different effector signalling pathways.
    Here, we analysed the distribution of GABAB receptors using highly sensitive SDS-digested
    freeze-fracture replica labelling in mouse cerebellar Purkinje cells. Immunoreactivity
    for GABAB1 was observed on presynaptic and, more abundantly, on postsynaptic compartments,
    showing both scattered and clustered distribution patterns. Quantitative analysis
    of immunoparticles revealed a somato-dendritic gradient, with the density of immunoparticles
    increasing 26-fold from somata to dendritic spines. To understand the spatial
    relationship of GABAB receptors with two key effector ion channels, the G protein-gated
    inwardly rectifying K+ (GIRK/Kir3) channel and the voltage-dependent Ca2+ channel,
    biochemical and immunohistochemical approaches were performed. Co-immunoprecipitation
    analysis demonstrated that GABAB receptors co-assembled with GIRK and CaV2.1 channels
    in the cerebellum. Using double-labelling immunoelectron microscopic techniques,
    co-clustering between GABAB1 and GIRK2 was detected in dendritic spines, whereas
    they were mainly segregated in the dendritic shafts. In contrast, co-clustering
    of GABAB1 and CaV2.1 was detected in dendritic shafts but not spines. Presynaptically,
    although no significant co-clustering of GABAB1 and GIRK2 or CaV2.1 channels was
    detected, inter-cluster distance for GABAB1 and GIRK2 was significantly smaller
    in the active zone than in the dendritic shafts, and that for GABAB1 and CaV2.1
    was significantly smaller in the active zone than in the dendritic shafts and
    spines. Thus, GABAB receptors are associated with GIRK and CaV2.1 channels in
    different subcellular compartments. These data provide a better framework for
    understanding the different roles played by GABAB receptors and their effector
    ion channels in the cerebellar network.
article_processing_charge: No
article_type: original
author:
- first_name: Rafael
  full_name: Luján, Rafael
  last_name: Luján
- first_name: Carolina
  full_name: Aguado, Carolina
  last_name: Aguado
- first_name: Francisco
  full_name: Ciruela, Francisco
  last_name: Ciruela
- first_name: Javier
  full_name: Cózar, Javier
  last_name: Cózar
- first_name: David
  full_name: Kleindienst, David
  id: 42E121A4-F248-11E8-B48F-1D18A9856A87
  last_name: Kleindienst
- first_name: Luis
  full_name: De La Ossa, Luis
  last_name: De La Ossa
- first_name: Bernhard
  full_name: Bettler, Bernhard
  last_name: Bettler
- first_name: Kevin
  full_name: Wickman, Kevin
  last_name: Wickman
- first_name: Masahiko
  full_name: Watanabe, Masahiko
  last_name: Watanabe
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
citation:
  ama: Luján R, Aguado C, Ciruela F, et al. Differential association of GABAB receptors
    with their effector ion channels in Purkinje cells. <i>Brain Structure and Function</i>.
    2018;223(3):1565-1587. doi:<a href="https://doi.org/10.1007/s00429-017-1568-y">10.1007/s00429-017-1568-y</a>
  apa: Luján, R., Aguado, C., Ciruela, F., Cózar, J., Kleindienst, D., De La Ossa,
    L., … Fukazawa, Y. (2018). Differential association of GABAB receptors with their
    effector ion channels in Purkinje cells. <i>Brain Structure and Function</i>.
    Springer. <a href="https://doi.org/10.1007/s00429-017-1568-y">https://doi.org/10.1007/s00429-017-1568-y</a>
  chicago: Luján, Rafael, Carolina Aguado, Francisco Ciruela, Javier Cózar, David
    Kleindienst, Luis De La Ossa, Bernhard Bettler, et al. “Differential Association
    of GABAB Receptors with Their Effector Ion Channels in Purkinje Cells.” <i>Brain
    Structure and Function</i>. Springer, 2018. <a href="https://doi.org/10.1007/s00429-017-1568-y">https://doi.org/10.1007/s00429-017-1568-y</a>.
  ieee: R. Luján <i>et al.</i>, “Differential association of GABAB receptors with
    their effector ion channels in Purkinje cells,” <i>Brain Structure and Function</i>,
    vol. 223, no. 3. Springer, pp. 1565–1587, 2018.
  ista: Luján R, Aguado C, Ciruela F, Cózar J, Kleindienst D, De La Ossa L, Bettler
    B, Wickman K, Watanabe M, Shigemoto R, Fukazawa Y. 2018. Differential association
    of GABAB receptors with their effector ion channels in Purkinje cells. Brain Structure
    and Function. 223(3), 1565–1587.
  mla: Luján, Rafael, et al. “Differential Association of GABAB Receptors with Their
    Effector Ion Channels in Purkinje Cells.” <i>Brain Structure and Function</i>,
    vol. 223, no. 3, Springer, 2018, pp. 1565–87, doi:<a href="https://doi.org/10.1007/s00429-017-1568-y">10.1007/s00429-017-1568-y</a>.
  short: R. Luján, C. Aguado, F. Ciruela, J. Cózar, D. Kleindienst, L. De La Ossa,
    B. Bettler, K. Wickman, M. Watanabe, R. Shigemoto, Y. Fukazawa, Brain Structure
    and Function 223 (2018) 1565–1587.
date_created: 2018-12-11T11:47:29Z
date_published: 2018-04-01T00:00:00Z
date_updated: 2024-03-25T23:30:16Z
day: '01'
ddc:
- '571'
department:
- _id: RySh
doi: 10.1007/s00429-017-1568-y
ec_funded: 1
external_id:
  isi:
  - '000428419500030'
file:
- access_level: open_access
  checksum: a55b3103476ecb5f4f983d8801807e8b
  content_type: application/pdf
  creator: system
  date_created: 2018-12-12T10:15:36Z
  date_updated: 2020-07-14T12:47:20Z
  file_id: '5157'
  file_name: IST-2018-1013-v1+1_2018_Kleindienst_Differential.pdf
  file_size: 5542926
  relation: main_file
file_date_updated: 2020-07-14T12:47:20Z
has_accepted_license: '1'
intvolume: '       223'
isi: 1
issue: '3'
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
page: 1565 - 1587
project:
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
- _id: 25681D80-B435-11E9-9278-68D0E5697425
  call_identifier: FP7
  grant_number: '291734'
  name: International IST Postdoc Fellowship Programme
publication: Brain Structure and Function
publication_status: published
publisher: Springer
publist_id: '7192'
pubrep_id: '1013'
quality_controlled: '1'
related_material:
  record:
  - id: '9562'
    relation: dissertation_contains
    status: public
scopus_import: '1'
status: public
title: Differential association of GABAB receptors with their effector ion channels
  in Purkinje 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: 223
year: '2018'
...
---
_id: '41'
abstract:
- lang: eng
  text: 'The small-conductance, Ca2+-activated K+ (SK) channel subtype SK2 regulates
    the spike rate and firing frequency, as well as Ca2+ transients in Purkinje cells
    (PCs). To understand the molecular basis by which SK2 channels mediate these functions,
    we analyzed the exact location and densities of SK2 channels along the neuronal
    surface of the mouse cerebellar PCs using SDS-digested freeze-fracture replica
    labeling (SDS-FRL) of high sensitivity combined with quantitative analyses. Immunogold
    particles for SK2 were observed on post- and pre-synaptic compartments showing
    both scattered and clustered distribution patterns. We found an axo-somato-dendritic
    gradient of the SK2 particle density increasing 12-fold from soma to dendritic
    spines. Using two different immunogold approaches, we also found that SK2 immunoparticles
    were frequently adjacent to, but never overlap with, the postsynaptic density
    of excitatory synapses in PC spines. Co-immunoprecipitation analysis demonstrated
    that SK2 channels form macromolecular complexes with two types of proteins that
    mobilize Ca2+: CaV2.1 channels and mGlu1α receptors in the cerebellum. Freeze-fracture
    replica double-labeling showed significant co-clustering of particles for SK2
    with those for CaV2.1 channels and mGlu1α receptors. SK2 channels were also detected
    at presynaptic sites, mostly at the presynaptic active zone (AZ), where they are
    close to CaV2.1 channels, though they are not significantly co-clustered. These
    data demonstrate that SK2 channels located in different neuronal compartments
    can associate with distinct proteins mobilizing Ca2+, and suggest that the ultrastructural
    association of SK2 with CaV2.1 and mGlu1α provides the mechanism that ensures
    voltage (excitability) regulation by distinct intracellular Ca2+ transients in
    PCs.'
article_number: '311'
article_processing_charge: No
article_type: original
author:
- first_name: Rafæl
  full_name: Luján, Rafæl
  last_name: Luján
- first_name: Carolina
  full_name: Aguado, Carolina
  last_name: Aguado
- first_name: Francisco
  full_name: Ciruela, Francisco
  last_name: Ciruela
- first_name: Xavier
  full_name: Arus, Xavier
  last_name: Arus
- first_name: Alejandro
  full_name: Martín Belmonte, Alejandro
  last_name: Martín Belmonte
- first_name: Rocío
  full_name: Alfaro Ruiz, Rocío
  last_name: Alfaro Ruiz
- first_name: Jesus
  full_name: Martinez Gomez, Jesus
  last_name: Martinez Gomez
- first_name: Luis
  full_name: De La Ossa, Luis
  last_name: De La Ossa
- first_name: Masahiko
  full_name: Watanabe, Masahiko
  last_name: Watanabe
- first_name: John
  full_name: Adelman, John
  last_name: Adelman
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
citation:
  ama: Luján R, Aguado C, Ciruela F, et al. Sk2 channels associate with mGlu1α receptors
    and CaV2.1 channels in Purkinje cells. <i>Frontiers in Cellular Neuroscience</i>.
    2018;12. doi:<a href="https://doi.org/10.3389/fncel.2018.00311">10.3389/fncel.2018.00311</a>
  apa: Luján, R., Aguado, C., Ciruela, F., Arus, X., Martín Belmonte, A., Alfaro Ruiz,
    R., … Fukazawa, Y. (2018). Sk2 channels associate with mGlu1α receptors and CaV2.1
    channels in Purkinje cells. <i>Frontiers in Cellular Neuroscience</i>. Frontiers
    Media. <a href="https://doi.org/10.3389/fncel.2018.00311">https://doi.org/10.3389/fncel.2018.00311</a>
  chicago: Luján, Rafæl, Carolina Aguado, Francisco Ciruela, Xavier Arus, Alejandro
    Martín Belmonte, Rocío Alfaro Ruiz, Jesus Martinez Gomez, et al. “Sk2 Channels
    Associate with MGlu1α Receptors and CaV2.1 Channels in Purkinje Cells.” <i>Frontiers
    in Cellular Neuroscience</i>. Frontiers Media, 2018. <a href="https://doi.org/10.3389/fncel.2018.00311">https://doi.org/10.3389/fncel.2018.00311</a>.
  ieee: R. Luján <i>et al.</i>, “Sk2 channels associate with mGlu1α receptors and
    CaV2.1 channels in Purkinje cells,” <i>Frontiers in Cellular Neuroscience</i>,
    vol. 12. Frontiers Media, 2018.
  ista: Luján R, Aguado C, Ciruela F, Arus X, Martín Belmonte A, Alfaro Ruiz R, Martinez
    Gomez J, De La Ossa L, Watanabe M, Adelman J, Shigemoto R, Fukazawa Y. 2018. Sk2
    channels associate with mGlu1α receptors and CaV2.1 channels in Purkinje cells.
    Frontiers in Cellular Neuroscience. 12, 311.
  mla: Luján, Rafæl, et al. “Sk2 Channels Associate with MGlu1α Receptors and CaV2.1
    Channels in Purkinje Cells.” <i>Frontiers in Cellular Neuroscience</i>, vol. 12,
    311, Frontiers Media, 2018, doi:<a href="https://doi.org/10.3389/fncel.2018.00311">10.3389/fncel.2018.00311</a>.
  short: R. Luján, C. Aguado, F. Ciruela, X. Arus, A. Martín Belmonte, R. Alfaro Ruiz,
    J. Martinez Gomez, L. De La Ossa, M. Watanabe, J. Adelman, R. Shigemoto, Y. Fukazawa,
    Frontiers in Cellular Neuroscience 12 (2018).
date_created: 2018-12-11T11:44:19Z
date_published: 2018-09-19T00:00:00Z
date_updated: 2023-09-18T09:31:18Z
day: '19'
ddc:
- '570'
department:
- _id: RySh
doi: 10.3389/fncel.2018.00311
ec_funded: 1
external_id:
  isi:
  - '000445090100002'
file:
- access_level: open_access
  checksum: 0bcaec8d596162af0b7fe3f31325d480
  content_type: application/pdf
  creator: dernst
  date_created: 2018-12-17T08:49:03Z
  date_updated: 2020-07-14T12:46:23Z
  file_id: '5684'
  file_name: fncel-12-00311.pdf
  file_size: 6834251
  relation: main_file
file_date_updated: 2020-07-14T12:46:23Z
has_accepted_license: '1'
intvolume: '        12'
isi: 1
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
project:
- _id: 25CBA828-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '720270'
  name: Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)
publication: Frontiers in Cellular Neuroscience
publication_identifier:
  issn:
  - '16625102'
publication_status: published
publisher: Frontiers Media
publist_id: '8013'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Sk2 channels associate with mGlu1α receptors and CaV2.1 channels in Purkinje
  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: 12
year: '2018'
...