[{"volume":169,"publication_status":"published","date_published":"2021-07-27T00:00:00Z","_id":"9756","abstract":[{"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.","lang":"eng"}],"article_processing_charge":"No","publication_identifier":{"isbn":["9781071615218"],"eisbn":["9781071615225"]},"user_id":"D865714E-FA4E-11E9-B85B-F5C5E5697425","date_updated":"2024-03-25T23:30:16Z","has_accepted_license":"1","year":"2021","oa_version":"None","status":"public","intvolume":"       169","quality_controlled":"1","department":[{"_id":"RySh"},{"_id":"EM-Fac"}],"publication":" Receptor and Ion Channel Detection in the Brain","publisher":"Humana","date_created":"2021-07-30T09:34:56Z","series_title":"Neuromethods","place":"New York","month":"07","page":"267-283","ddc":["573"],"doi":"10.1007/978-1-0716-1522-5_19","language":[{"iso":"eng"}],"keyword":["Freeze-fracture replica: Deep learning","Immunogold labeling","Integral membrane protein","Electron microscopy"],"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.).","project":[{"call_identifier":"H2020","_id":"25CA28EA-B435-11E9-9278-68D0E5697425","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","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)","grant_number":"720270"}],"related_material":{"record":[{"id":"9562","relation":"dissertation_contains","status":"public"}]},"alternative_title":["Neuromethods"],"type":"book_chapter","author":[{"id":"3F99E422-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-9735-5315","last_name":"Kaufmann","full_name":"Kaufmann, Walter","first_name":"Walter"},{"first_name":"David","full_name":"Kleindienst, David","last_name":"Kleindienst","id":"42E121A4-F248-11E8-B48F-1D18A9856A87"},{"id":"2E55CDF2-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-7429-7896","last_name":"Harada","full_name":"Harada, Harumi","first_name":"Harumi"},{"id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444","last_name":"Shigemoto","first_name":"Ryuichi","full_name":"Shigemoto, Ryuichi"}],"day":"27","title":"High-Resolution localization and quantitation of membrane proteins by SDS-digested freeze-fracture replica labeling (SDS-FRL)","ec_funded":1,"citation":{"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.","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>","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>","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.","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.","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>.","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>."}},{"project":[{"grant_number":"720270","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)","_id":"25CBA828-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"},{"call_identifier":"H2020","_id":"26436750-B435-11E9-9278-68D0E5697425","grant_number":"785907","name":"Human Brain Project Specific Grant Agreement 2 (HBP SGA 2)"}],"pmid":1,"ddc":["570"],"doi":"10.1111/bpa.12802","language":[{"iso":"eng"}],"title":"Reduction in the neuronal surface of post and presynaptic GABA>B< receptors in the hippocampus in a mouse model of Alzheimer's disease","ec_funded":1,"citation":{"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.","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.","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>.","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.","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>","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>"},"author":[{"last_name":"Martín-Belmonte","full_name":"Martín-Belmonte, Alejandro","first_name":"Alejandro"},{"first_name":"Carolina","full_name":"Aguado, Carolina","last_name":"Aguado"},{"last_name":"Alfaro-Ruíz","first_name":"Rocío","full_name":"Alfaro-Ruíz, Rocío"},{"last_name":"Moreno-Martínez","full_name":"Moreno-Martínez, Ana Esther","first_name":"Ana Esther"},{"last_name":"De La Ossa","first_name":"Luis","full_name":"De La Ossa, Luis"},{"full_name":"Martínez-Hernández, José","first_name":"José","last_name":"Martínez-Hernández"},{"full_name":"Buisson, Alain","first_name":"Alain","last_name":"Buisson"},{"full_name":"Früh, Simon","first_name":"Simon","last_name":"Früh"},{"last_name":"Bettler","full_name":"Bettler, Bernhard","first_name":"Bernhard"},{"id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444","first_name":"Ryuichi","full_name":"Shigemoto, Ryuichi","last_name":"Shigemoto"},{"last_name":"Fukazawa","first_name":"Yugo","full_name":"Fukazawa, Yugo"},{"last_name":"Luján","first_name":"Rafael","full_name":"Luján, Rafael"}],"type":"journal_article","day":"01","isi":1,"publisher":"Wiley","status":"public","intvolume":"        30","quality_controlled":"1","department":[{"_id":"RySh"}],"publication":"Brain Pathology","page":"554-575","date_created":"2019-12-22T23:00:43Z","month":"05","publication_identifier":{"eissn":["17503639"],"issn":["10156305"]},"date_updated":"2023-09-06T14:48:01Z","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","scopus_import":"1","external_id":{"isi":["000502270900001"],"pmid":["31729777"]},"has_accepted_license":"1","oa_version":"Published Version","year":"2020","article_type":"original","oa":1,"publication_status":"published","volume":30,"tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)"},"file_date_updated":"2020-09-22T09:47:19Z","issue":"3","article_processing_charge":"No","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."}],"_id":"7207","date_published":"2020-05-01T00:00:00Z","file":[{"date_created":"2020-09-22T09:47:19Z","success":1,"content_type":"application/pdf","relation":"main_file","file_id":"8554","checksum":"549cc1b18f638a21d17a939ba5563fa9","date_updated":"2020-09-22T09:47:19Z","access_level":"open_access","file_name":"2020_BrainPathology_MartinBelmonte.pdf","creator":"dernst","file_size":4220935}]},{"ddc":["570"],"doi":"10.1016/j.isci.2019.11.025","language":[{"iso":"eng"}],"project":[{"grant_number":"694539","name":"In situ analysis of single channel subunit composition in neurons: physiological implication in synaptic plasticity and behaviour","call_identifier":"H2020","_id":"25CA28EA-B435-11E9-9278-68D0E5697425"},{"call_identifier":"H2020","_id":"25CBA828-B435-11E9-9278-68D0E5697425","grant_number":"720270","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)"}],"pmid":1,"related_material":{"record":[{"relation":"dissertation_contains","id":"11393","status":"public"}]},"author":[{"id":"4427179E-F248-11E8-B48F-1D18A9856A87","first_name":"Shigekazu","full_name":"Tabata, Shigekazu","last_name":"Tabata"},{"id":"4BE3BC94-F248-11E8-B48F-1D18A9856A87","last_name":"Jevtic","first_name":"Marijo","full_name":"Jevtic, Marijo"},{"first_name":"Nobutaka","full_name":"Kurashige, Nobutaka","last_name":"Kurashige"},{"last_name":"Fuchida","full_name":"Fuchida, Hirokazu","first_name":"Hirokazu"},{"last_name":"Kido","first_name":"Munetsugu","full_name":"Kido, Munetsugu"},{"last_name":"Tani","full_name":"Tani, Kazushi","first_name":"Kazushi"},{"first_name":"Naoki","full_name":"Zenmyo, Naoki","last_name":"Zenmyo"},{"full_name":"Uchinomiya, Shohei","first_name":"Shohei","last_name":"Uchinomiya"},{"first_name":"Harumi","full_name":"Harada, Harumi","last_name":"Harada","orcid":"0000-0001-7429-7896","id":"2E55CDF2-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Itakura","full_name":"Itakura, Makoto","first_name":"Makoto"},{"full_name":"Hamachi, Itaru","first_name":"Itaru","last_name":"Hamachi"},{"full_name":"Shigemoto, Ryuichi","first_name":"Ryuichi","last_name":"Shigemoto","orcid":"0000-0001-8761-9444","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Ojida","full_name":"Ojida, Akio","first_name":"Akio"}],"type":"journal_article","day":"20","title":"Electron microscopic detection of single membrane proteins by a specific chemical labeling","ec_funded":1,"citation":{"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.","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>"},"intvolume":"        22","status":"public","quality_controlled":"1","department":[{"_id":"RySh"}],"publication":"iScience","publisher":"Elsevier","date_created":"2020-01-29T15:56:56Z","month":"12","page":"256-268","publication_identifier":{"issn":["2589-0042"]},"user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","date_updated":"2024-03-25T23:30:07Z","external_id":{"isi":[":000504652000020"],"pmid":["31786521"]},"scopus_import":"1","year":"2019","has_accepted_license":"1","oa_version":"Published Version","article_type":"original","volume":22,"tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)"},"file_date_updated":"2020-07-14T12:47:57Z","oa":1,"publication_status":"published","_id":"7391","date_published":"2019-12-20T00:00:00Z","abstract":[{"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.","lang":"eng"}],"file":[{"file_name":"2019_iScience_Tabata.pdf","creator":"dernst","file_size":7197776,"checksum":"f3e90056a49f09b205b1c4f8c739ffd1","date_updated":"2020-07-14T12:47:57Z","access_level":"open_access","content_type":"application/pdf","relation":"main_file","file_id":"7448","date_created":"2020-02-04T10:48:36Z"}],"issue":"12","article_processing_charge":"No"},{"issue":"5","article_processing_charge":"Yes","_id":"292","date_published":"2018-05-10T00:00:00Z","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."}],"file":[{"file_name":"2018_Plos_Botella_Soler.pdf","file_size":3460786,"creator":"dernst","date_updated":"2020-07-14T12:45:53Z","access_level":"open_access","checksum":"3026f94d235219e15514505fdbadf34e","content_type":"application/pdf","relation":"main_file","file_id":"5974","date_created":"2019-02-13T11:07:15Z"}],"article_number":"e1006057","oa":1,"publication_status":"published","volume":14,"tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)"},"file_date_updated":"2020-07-14T12:45:53Z","oa_version":"Published Version","has_accepted_license":"1","year":"2018","article_type":"original","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","date_updated":"2024-02-21T13:45:25Z","scopus_import":"1","external_id":{"isi":["000434012100002"]},"date_created":"2018-12-11T11:45:39Z","month":"05","isi":1,"publisher":"Public Library of Science","status":"public","intvolume":"        14","quality_controlled":"1","department":[{"_id":"GaTk"}],"publication":"PLoS Computational Biology","title":"Nonlinear decoding of a complex movie from the mammalian retina","ec_funded":1,"citation":{"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>","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>","short":"V. Botella Soler, S. Deny, G.S. Martius, O. Marre, G. Tkačik, PLoS Computational Biology 14 (2018).","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>.","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.","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>."},"type":"journal_article","author":[{"id":"421234E8-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8790-1914","last_name":"Botella Soler","first_name":"Vicent","full_name":"Botella Soler, Vicent"},{"last_name":"Deny","first_name":"Stephane","full_name":"Deny, Stephane"},{"first_name":"Georg S","full_name":"Martius, Georg S","last_name":"Martius"},{"last_name":"Marre","first_name":"Olivier","full_name":"Marre, Olivier"},{"last_name":"Tkacik","full_name":"Tkacik, Gasper","first_name":"Gasper","id":"3D494DCA-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-6699-1455"}],"day":"10","project":[{"call_identifier":"H2020","_id":"25CBA828-B435-11E9-9278-68D0E5697425","grant_number":"720270","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)"},{"name":"Sensitivity to higher-order statistics in natural scenes","grant_number":"P 25651-N26","_id":"254D1A94-B435-11E9-9278-68D0E5697425","call_identifier":"FWF"}],"related_material":{"link":[{"url":"https://ist.ac.at/en/news/video-of-moving-discs-reconstructed-from-rat-retinal-neuron-signals/","description":"News on IST Homepage","relation":"press_release"}],"record":[{"status":"public","id":"5584","relation":"research_data"}]},"ddc":["570"],"doi":"10.1371/journal.pcbi.1006057","language":[{"iso":"eng"}]},{"status":"public","intvolume":"       223","quality_controlled":"1","department":[{"_id":"RySh"}],"publication":"Brain Structure and Function","isi":1,"publisher":"Springer","date_created":"2018-12-11T11:47:29Z","month":"04","page":"1565 - 1587","doi":"10.1007/s00429-017-1568-y","ddc":["571"],"language":[{"iso":"eng"}],"related_material":{"record":[{"id":"9562","relation":"dissertation_contains","status":"public"}]},"project":[{"call_identifier":"H2020","_id":"25CBA828-B435-11E9-9278-68D0E5697425","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)","grant_number":"720270"},{"_id":"25681D80-B435-11E9-9278-68D0E5697425","call_identifier":"FP7","name":"International IST Postdoc Fellowship Programme","grant_number":"291734"}],"type":"journal_article","author":[{"full_name":"Luján, Rafael","first_name":"Rafael","last_name":"Luján"},{"first_name":"Carolina","full_name":"Aguado, Carolina","last_name":"Aguado"},{"first_name":"Francisco","full_name":"Ciruela, Francisco","last_name":"Ciruela"},{"full_name":"Cózar, Javier","first_name":"Javier","last_name":"Cózar"},{"first_name":"David","full_name":"Kleindienst, David","last_name":"Kleindienst","id":"42E121A4-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Luis","full_name":"De La Ossa, Luis","last_name":"De La Ossa"},{"last_name":"Bettler","full_name":"Bettler, Bernhard","first_name":"Bernhard"},{"first_name":"Kevin","full_name":"Wickman, Kevin","last_name":"Wickman"},{"last_name":"Watanabe","full_name":"Watanabe, Masahiko","first_name":"Masahiko"},{"first_name":"Ryuichi","full_name":"Shigemoto, Ryuichi","last_name":"Shigemoto","orcid":"0000-0001-8761-9444","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Fukazawa","full_name":"Fukazawa, Yugo","first_name":"Yugo"}],"day":"01","title":"Differential association of GABAB receptors with their effector ion channels in Purkinje cells","ec_funded":1,"citation":{"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.","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>.","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.","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>","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>"},"volume":223,"pubrep_id":"1013","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)"},"file_date_updated":"2020-07-14T12:47:20Z","oa":1,"publication_status":"published","_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."}],"date_published":"2018-04-01T00:00:00Z","file":[{"content_type":"application/pdf","file_id":"5157","relation":"main_file","date_created":"2018-12-12T10:15:36Z","file_name":"IST-2018-1013-v1+1_2018_Kleindienst_Differential.pdf","creator":"system","file_size":5542926,"date_updated":"2020-07-14T12:47:20Z","access_level":"open_access","checksum":"a55b3103476ecb5f4f983d8801807e8b"}],"issue":"3","article_processing_charge":"No","date_updated":"2024-03-25T23:30:16Z","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","scopus_import":"1","external_id":{"isi":["000428419500030"]},"has_accepted_license":"1","oa_version":"Published Version","year":"2018","publist_id":"7192","article_type":"original"},{"date_created":"2018-12-11T11:44:19Z","month":"09","intvolume":"        12","status":"public","publication":"Frontiers in Cellular Neuroscience","quality_controlled":"1","department":[{"_id":"RySh"}],"isi":1,"publisher":"Frontiers Media","type":"journal_article","author":[{"full_name":"Luján, Rafæl","first_name":"Rafæl","last_name":"Luján"},{"last_name":"Aguado","full_name":"Aguado, Carolina","first_name":"Carolina"},{"first_name":"Francisco","full_name":"Ciruela, Francisco","last_name":"Ciruela"},{"last_name":"Arus","first_name":"Xavier","full_name":"Arus, Xavier"},{"last_name":"Martín Belmonte","first_name":"Alejandro","full_name":"Martín Belmonte, Alejandro"},{"first_name":"Rocío","full_name":"Alfaro Ruiz, Rocío","last_name":"Alfaro Ruiz"},{"first_name":"Jesus","full_name":"Martinez Gomez, Jesus","last_name":"Martinez Gomez"},{"full_name":"De La Ossa, Luis","first_name":"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"},{"last_name":"Shigemoto","full_name":"Shigemoto, Ryuichi","first_name":"Ryuichi","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444"},{"last_name":"Fukazawa","full_name":"Fukazawa, Yugo","first_name":"Yugo"}],"day":"19","title":"Sk2 channels associate with mGlu1α receptors and CaV2.1 channels in Purkinje cells","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>","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).","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>.","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>.","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.","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."},"ec_funded":1,"doi":"10.3389/fncel.2018.00311","ddc":["570"],"language":[{"iso":"eng"}],"project":[{"_id":"25CBA828-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","grant_number":"720270","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)"}],"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."}],"_id":"41","date_published":"2018-09-19T00:00:00Z","file":[{"content_type":"application/pdf","file_id":"5684","relation":"main_file","date_created":"2018-12-17T08:49:03Z","file_name":"fncel-12-00311.pdf","file_size":6834251,"creator":"dernst","checksum":"0bcaec8d596162af0b7fe3f31325d480","date_updated":"2020-07-14T12:46:23Z","access_level":"open_access"}],"article_number":"311","article_processing_charge":"No","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)"},"volume":12,"file_date_updated":"2020-07-14T12:46:23Z","oa":1,"publication_status":"published","has_accepted_license":"1","year":"2018","oa_version":"Published Version","article_type":"original","publist_id":"8013","publication_identifier":{"issn":["16625102"]},"external_id":{"isi":["000445090100002"]},"scopus_import":"1","date_updated":"2023-09-18T09:31:18Z","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1"}]