[{"ec_funded":1,"issue":"Supplement_1","volume":71,"publication_status":"published","publication_identifier":{"issn":["2050-5698"],"eissn":["2050-5701"]},"language":[{"iso":"eng"}],"main_file_link":[{"url":"https://doi.org/10.1093/jmicro/dfab048","open_access":"1"}],"scopus_import":"1","intvolume":" 71","month":"03","abstract":[{"lang":"eng","text":"Genetically encoded tags have introduced extensive lines of application from purification of tagged proteins to their visualization at the single molecular, cellular, histological and whole-body levels. Combined with other rapidly developing technologies such as clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, proteomics, super-resolution microscopy and proximity labeling, a large variety of genetically encoded tags have been developed in the last two decades. In this review, I focus on the current status of tag development for electron microscopic (EM) visualization of proteins with metal particle labeling. Compared with conventional immunoelectron microscopy using gold particles, tag-mediated metal particle labeling has several advantages that could potentially improve the sensitivity, spatial and temporal resolution, and applicability to a wide range of proteins of interest (POIs). It may enable researchers to detect single molecules in situ, allowing the quantitative measurement of absolute numbers and exact localization patterns of POI in the ultrastructural context. Thus, genetically encoded tags for EM could revolutionize the field as green fluorescence protein did for light microscopy, although we still have many challenges to overcome before reaching this goal."}],"pmid":1,"oa_version":"Published Version","department":[{"_id":"RySh"}],"date_updated":"2023-08-03T06:08:01Z","article_type":"original","type":"journal_article","status":"public","_id":"10889","page":"i72-i80","date_created":"2022-03-20T23:01:39Z","date_published":"2022-03-01T00:00:00Z","doi":"10.1093/jmicro/dfab048","year":"2022","isi":1,"publication":"Microscopy","day":"01","oa":1,"quality_controlled":"1","publisher":"Oxford Academic","acknowledgement":"European Research Council Advanced Grant (694539 to R.S.).","article_processing_charge":"No","external_id":{"isi":["000768384100011"],"pmid":["35275179"]},"author":[{"last_name":"Shigemoto","full_name":"Shigemoto, Ryuichi","orcid":"0000-0001-8761-9444","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","first_name":"Ryuichi"}],"title":"Electron microscopic visualization of single molecules by tag-mediated metal particle labeling","citation":{"short":"R. Shigemoto, Microscopy 71 (2022) i72–i80.","ieee":"R. Shigemoto, “Electron microscopic visualization of single molecules by tag-mediated metal particle labeling,” Microscopy, vol. 71, no. Supplement_1. Oxford Academic, pp. i72–i80, 2022.","apa":"Shigemoto, R. (2022). Electron microscopic visualization of single molecules by tag-mediated metal particle labeling. Microscopy. Oxford Academic. https://doi.org/10.1093/jmicro/dfab048","ama":"Shigemoto R. Electron microscopic visualization of single molecules by tag-mediated metal particle labeling. Microscopy. 2022;71(Supplement_1):i72-i80. doi:10.1093/jmicro/dfab048","mla":"Shigemoto, Ryuichi. “Electron Microscopic Visualization of Single Molecules by Tag-Mediated Metal Particle Labeling.” Microscopy, vol. 71, no. Supplement_1, Oxford Academic, 2022, pp. i72–80, doi:10.1093/jmicro/dfab048.","ista":"Shigemoto R. 2022. Electron microscopic visualization of single molecules by tag-mediated metal particle labeling. Microscopy. 71(Supplement_1), i72–i80.","chicago":"Shigemoto, Ryuichi. “Electron Microscopic Visualization of Single Molecules by Tag-Mediated Metal Particle Labeling.” Microscopy. Oxford Academic, 2022. https://doi.org/10.1093/jmicro/dfab048."},"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","project":[{"grant_number":"694539","name":"In situ analysis of single channel subunit composition in neurons: physiological implication in synaptic plasticity and behaviour","_id":"25CA28EA-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"}]},{"abstract":[{"lang":"eng","text":"Elevation of soluble wild-type (WT) tau occurs in synaptic compartments in Alzheimer’s disease. We addressed whether tau elevation affects synaptic transmission at the calyx of Held in slices from mice brainstem. Whole-cell loading of WT human tau (h-tau) in presynaptic terminals at 10–20 µM caused microtubule (MT) assembly and activity-dependent rundown of excitatory neurotransmission. Capacitance measurements revealed that the primary target of WT h-tau is vesicle endocytosis. Blocking MT assembly using nocodazole prevented tau-induced impairments of endocytosis and neurotransmission. Immunofluorescence imaging analyses revealed that MT assembly by WT h-tau loading was associated with an increased MT-bound fraction of the endocytic protein dynamin. A synthetic dodecapeptide corresponding to dynamin 1-pleckstrin-homology domain inhibited MT-dynamin interaction and rescued tau-induced impairments of endocytosis and neurotransmission. We conclude that elevation of presynaptic WT tau induces de novo assembly of MTs, thereby sequestering free dynamins. As a result, endocytosis and subsequent vesicle replenishment are impaired, causing activity-dependent rundown of neurotransmission."}],"pmid":1,"oa_version":"Published Version","scopus_import":"1","intvolume":" 11","month":"05","publication_status":"published","publication_identifier":{"eissn":["2050-084X"]},"language":[{"iso":"eng"}],"file":[{"content_type":"application/pdf","relation":"main_file","access_level":"open_access","success":1,"file_id":"11421","checksum":"ccddbd167e00ff8375f12998af497152","file_size":2466296,"date_updated":"2022-05-30T08:09:16Z","creator":"cchlebak","file_name":"elife-73542-v2.pdf","date_created":"2022-05-30T08:09:16Z"}],"volume":11,"_id":"11419","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"},"article_type":"original","type":"journal_article","status":"public","date_updated":"2023-08-03T07:15:49Z","ddc":["616"],"file_date_updated":"2022-05-30T08:09:16Z","department":[{"_id":"RySh"}],"acknowledgement":"We thank Yasuo Ihara, Nobuyuki Nukina, and Takeshi Sakaba for comments and Patrick Stoney for editing this paper. We also thank Shota Okuda and Mikako Matsubara for their contributions in the early stage of this study, and Satoko Wada-Kakuda for technical assistant with in vitro analysis of tau. This research was supported by funding from Okinawa Institute of Science and Technology and from Technology (OIST) and Core Research for the Evolutional Science and Technology of Japan Science and Technology Agency (CREST) to TT, and by Scientific Research on Innovative Areas to TM (Brain Protein Aging and Dementia Control 26117004).","oa":1,"publisher":"eLife Sciences Publications","quality_controlled":"1","year":"2022","isi":1,"has_accepted_license":"1","publication":"eLife","day":"05","date_created":"2022-05-29T22:01:54Z","date_published":"2022-05-05T00:00:00Z","doi":"10.7554/eLife.73542","article_number":"e73542","citation":{"mla":"Hori, Tetsuya, et al. “Microtubule Assembly by Tau Impairs Endocytosis and Neurotransmission via Dynamin Sequestration in Alzheimer’s Disease Synapse Model.” ELife, vol. 11, e73542, eLife Sciences Publications, 2022, doi:10.7554/eLife.73542.","apa":"Hori, T., Eguchi, K., Wang, H. Y., Miyasaka, T., Guillaud, L., Taoufiq, Z., … Takahashi, T. (2022). Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer’s disease synapse model. ELife. eLife Sciences Publications. https://doi.org/10.7554/eLife.73542","ama":"Hori T, Eguchi K, Wang HY, et al. Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer’s disease synapse model. eLife. 2022;11. doi:10.7554/eLife.73542","ieee":"T. Hori et al., “Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer’s disease synapse model,” eLife, vol. 11. eLife Sciences Publications, 2022.","short":"T. Hori, K. Eguchi, H.Y. Wang, T. Miyasaka, L. Guillaud, Z. Taoufiq, S. Mahapatra, H. Yamada, K. Takei, T. Takahashi, ELife 11 (2022).","chicago":"Hori, Tetsuya, Kohgaku Eguchi, Han Ying Wang, Tomohiro Miyasaka, Laurent Guillaud, Zacharie Taoufiq, Satyajit Mahapatra, Hiroshi Yamada, Kohji Takei, and Tomoyuki Takahashi. “Microtubule Assembly by Tau Impairs Endocytosis and Neurotransmission via Dynamin Sequestration in Alzheimer’s Disease Synapse Model.” ELife. eLife Sciences Publications, 2022. https://doi.org/10.7554/eLife.73542.","ista":"Hori T, Eguchi K, Wang HY, Miyasaka T, Guillaud L, Taoufiq Z, Mahapatra S, Yamada H, Takei K, Takahashi T. 2022. Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer’s disease synapse model. eLife. 11, e73542."},"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","external_id":{"pmid":["35471147 "],"isi":["000876231600001"]},"article_processing_charge":"No","author":[{"first_name":"Tetsuya","last_name":"Hori","full_name":"Hori, Tetsuya"},{"full_name":"Eguchi, Kohgaku","orcid":"0000-0002-6170-2546","last_name":"Eguchi","first_name":"Kohgaku","id":"2B7846DC-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Wang, Han Ying","last_name":"Wang","first_name":"Han Ying"},{"first_name":"Tomohiro","last_name":"Miyasaka","full_name":"Miyasaka, Tomohiro"},{"last_name":"Guillaud","full_name":"Guillaud, Laurent","first_name":"Laurent"},{"last_name":"Taoufiq","full_name":"Taoufiq, Zacharie","first_name":"Zacharie"},{"first_name":"Satyajit","full_name":"Mahapatra, Satyajit","last_name":"Mahapatra"},{"first_name":"Hiroshi","full_name":"Yamada, Hiroshi","last_name":"Yamada"},{"full_name":"Takei, Kohji","last_name":"Takei","first_name":"Kohji"},{"last_name":"Takahashi","full_name":"Takahashi, Tomoyuki","first_name":"Tomoyuki"}],"title":"Microtubule assembly by tau impairs endocytosis and neurotransmission via dynamin sequestration in Alzheimer's disease synapse model"},{"language":[{"iso":"eng"}],"file":[{"content_type":"application/pdf","relation":"main_file","access_level":"open_access","success":1,"checksum":"88e49715ad6a1abf0fdb27efd65368dc","file_id":"12413","file_size":11013325,"date_updated":"2023-01-27T07:53:18Z","creator":"dernst","file_name":"2022_AlzheimersResearch_MartinBelmont.pdf","date_created":"2023-01-27T07:53:18Z"}],"publication_status":"published","publication_identifier":{"issn":["1758-9193"]},"volume":14,"oa_version":"Published Version","abstract":[{"lang":"eng","text":"Alzheimer’s disease (AD) is characterized by a reorganization of brain activity determining network hyperexcitability and loss of synaptic plasticity. Precisely, a dysfunction in metabotropic GABAB receptor signalling through G protein-gated inwardly rectifying K+ (GIRK or Kir3) channels on the hippocampus has been postulated. Thus, we determined the impact of amyloid-β (Aβ) pathology in GIRK channel density, subcellular distribution, and its association with GABAB receptors in hippocampal CA1 pyramidal neurons from the APP/PS1 mouse model using quantitative SDS-digested freeze-fracture replica labelling (SDS-FRL) and proximity ligation in situ assay (P-LISA). In wild type mice, single SDS-FRL detection revealed a similar dendritic gradient for GIRK1 and GIRK2 in CA1 pyramidal cells, with higher densities in spines, and GIRK3 showed a lower and uniform distribution. Double SDS-FRL showed a co-clustering of GIRK2 and GIRK1 in post- and presynaptic compartments, but not for GIRK2 and GIRK3. Likewise, double GABAB1 and GIRK2 SDS-FRL detection displayed a high degree of co-clustering in nanodomains (40–50 nm) mostly in spines and axon terminals. In APP/PS1 mice, the density of GIRK2 and GIRK1, but not for GIRK3, was significantly reduced along the neuronal surface of CA1 pyramidal cells and in axon terminals contacting them. Importantly, GABAB1 and GIRK2 co-clustering was not present in APP/PS1 mice. Similarly, P-LISA experiments revealed a significant reduction in GABAB1 and GIRK2 interaction on the hippocampus of this animal model. Overall, our results provide compelling evidence showing a significant reduction on the cell surface density of pre- and postsynaptic GIRK1 and GIRK2, but not GIRK3, and a decline in GABAB receptors and GIRK2 channels co-clustering in hippocampal pyramidal neurons from APP/PS1 mice, thus suggesting that a disruption in the GABAB receptor–GIRK channel membrane assembly causes dysregulation in the GABAB signalling via GIRK channels in this AD animal model."}],"intvolume":" 14","month":"09","scopus_import":"1","ddc":["570"],"date_updated":"2023-08-04T09:23:10Z","file_date_updated":"2023-01-27T07:53:18Z","department":[{"_id":"RySh"}],"_id":"12212","keyword":["Cognitive Neuroscience","Neurology (clinical)","Neurology"],"status":"public","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"},"article_type":"original","type":"journal_article","publication":"Alzheimer's Research & Therapy","day":"21","year":"2022","isi":1,"has_accepted_license":"1","date_created":"2023-01-16T09:45:51Z","date_published":"2022-09-21T00:00:00Z","doi":"10.1186/s13195-022-01078-5","acknowledgement":"We thank Ms. Diane Latawiec for the English revision of the manuscript. Funding sources were the Spanish Ministerio de Economía y Competitividad, Junta de Comunidades de Castilla-La Mancha (Spain), and Life Science Innovation Center at University of Fukui. We thank Centres de Recerca de Catalunya (CERCA) Programme/Generalitat de Catalunya for IDIBELL institutional support. We thank Hitoshi Takagi and Takako Maegawa at the University of Fukui for their technical assistance on SDS-FRL experiments.\r\nThis work was supported by grants from the Spanish Ministerio de Economía y Competitividad (BFU2015-63769-R, RTI2018-095812-B-I00, and PID2021-125875OB-I00) and Junta de Comunidades de Castilla-La Mancha (SBPLY/17/180501/000229 and SBPLY/21/180501/000064) to RL, Life Science Innovation Center at University of Fukui and JSPS KAKENHI (Grant Numbers 16H04662, 19H03323, and 20H05058) to YF, and Margarita Salas fellowship from Ministerio de Universidades and Universidad de Castilla-La Mancha to AMB.","oa":1,"publisher":"Springer Nature","quality_controlled":"1","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"mla":"Martín-Belmonte, Alejandro, et al. “Nanoscale Alterations in GABAB Receptors and GIRK Channel Organization on the Hippocampus of APP/PS1 Mice.” Alzheimer’s Research & Therapy, vol. 14, 136, Springer Nature, 2022, doi:10.1186/s13195-022-01078-5.","ama":"Martín-Belmonte A, Aguado C, Alfaro-Ruiz R, et al. Nanoscale alterations in GABAB receptors and GIRK channel organization on the hippocampus of APP/PS1 mice. Alzheimer’s Research & Therapy. 2022;14. doi:10.1186/s13195-022-01078-5","apa":"Martín-Belmonte, A., Aguado, C., Alfaro-Ruiz, R., Moreno-Martínez, A. E., de la Ossa, L., Aso, E., … Luján, R. (2022). Nanoscale alterations in GABAB receptors and GIRK channel organization on the hippocampus of APP/PS1 mice. Alzheimer’s Research & Therapy. Springer Nature. https://doi.org/10.1186/s13195-022-01078-5","short":"A. Martín-Belmonte, C. Aguado, R. Alfaro-Ruiz, A.E. Moreno-Martínez, L. de la Ossa, E. Aso, L. Gómez-Acero, R. Shigemoto, Y. Fukazawa, F. Ciruela, R. Luján, Alzheimer’s Research & Therapy 14 (2022).","ieee":"A. Martín-Belmonte et al., “Nanoscale alterations in GABAB receptors and GIRK channel organization on the hippocampus of APP/PS1 mice,” Alzheimer’s Research & Therapy, vol. 14. Springer Nature, 2022.","chicago":"Martín-Belmonte, Alejandro, Carolina Aguado, Rocío Alfaro-Ruiz, Ana Esther Moreno-Martínez, Luis de la Ossa, Ester Aso, Laura Gómez-Acero, et al. “Nanoscale Alterations in GABAB Receptors and GIRK Channel Organization on the Hippocampus of APP/PS1 Mice.” Alzheimer’s Research & Therapy. Springer Nature, 2022. https://doi.org/10.1186/s13195-022-01078-5.","ista":"Martín-Belmonte A, Aguado C, Alfaro-Ruiz R, Moreno-Martínez AE, de la Ossa L, Aso E, Gómez-Acero L, Shigemoto R, Fukazawa Y, Ciruela F, Luján R. 2022. Nanoscale alterations in GABAB receptors and GIRK channel organization on the hippocampus of APP/PS1 mice. Alzheimer’s Research & Therapy. 14, 136."},"title":"Nanoscale alterations in GABAB receptors and GIRK channel organization on the hippocampus of APP/PS1 mice","article_processing_charge":"No","external_id":{"isi":["000857985500001"]},"author":[{"first_name":"Alejandro","last_name":"Martín-Belmonte","full_name":"Martín-Belmonte, Alejandro"},{"last_name":"Aguado","full_name":"Aguado, Carolina","first_name":"Carolina"},{"first_name":"Rocío","full_name":"Alfaro-Ruiz, Rocío","last_name":"Alfaro-Ruiz"},{"first_name":"Ana Esther","full_name":"Moreno-Martínez, Ana Esther","last_name":"Moreno-Martínez"},{"last_name":"de la Ossa","full_name":"de la Ossa, Luis","first_name":"Luis"},{"last_name":"Aso","full_name":"Aso, Ester","first_name":"Ester"},{"first_name":"Laura","last_name":"Gómez-Acero","full_name":"Gómez-Acero, Laura"},{"last_name":"Shigemoto","orcid":"0000-0001-8761-9444","full_name":"Shigemoto, Ryuichi","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","first_name":"Ryuichi"},{"first_name":"Yugo","full_name":"Fukazawa, Yugo","last_name":"Fukazawa"},{"first_name":"Francisco","last_name":"Ciruela","full_name":"Ciruela, Francisco"},{"first_name":"Rafael","last_name":"Luján","full_name":"Luján, Rafael"}],"article_number":"136"},{"department":[{"_id":"RySh"}],"date_updated":"2023-10-03T10:58:31Z","status":"public","article_type":"original","type":"journal_article","_id":"11333","volume":28,"issue":"30","language":[{"iso":"eng"}],"publication_status":"published","publication_identifier":{"issn":["09476539"],"eissn":["15213765"]},"intvolume":" 28","month":"05","main_file_link":[{"open_access":"1","url":"https://doi.org/10.1002/chem.202200807"}],"scopus_import":"1","oa_version":"Published Version","pmid":1,"abstract":[{"lang":"eng","text":"Adenosine triphosphate (ATP) is the energy source for various biochemical processes and biomolecular motors in living things. Development of ATP antagonists and their stimuli-controlled actions offer a novel approach to regulate biological processes. Herein, we developed azobenzene-based photoswitchable ATP antagonists for controlling the activity of motor proteins; cytoplasmic and axonemal dyneins. The new ATP antagonists showed reversible photoswitching of cytoplasmic dynein activity in an in vitro dynein-microtubule system due to the trans and cis photoisomerization of their azobenzene segment. Importantly, our ATP antagonists reversibly regulated the axonemal dynein motor activity for the force generation in a demembranated model of Chlamydomonas reinhardtii. We found that the trans and cis isomers of ATP antagonists significantly differ in their affinity to the ATP binding site."}],"title":"Dynamic control of microbial movement by photoswitchable ATP antagonists","external_id":{"isi":["000781658800001"],"pmid":["35332959"]},"article_processing_charge":"No","author":[{"first_name":"Sampreeth","last_name":"Thayyil","full_name":"Thayyil, Sampreeth"},{"full_name":"Nishigami, Yukinori","last_name":"Nishigami","first_name":"Yukinori"},{"last_name":"Islam","full_name":"Islam, Muhammad J","id":"C94881D2-008F-11EA-8E08-2637E6697425","first_name":"Muhammad J"},{"last_name":"Hashim","full_name":"Hashim, P. K.","first_name":"P. K."},{"last_name":"Furuta","full_name":"Furuta, Ken'Ya","first_name":"Ken'Ya"},{"full_name":"Oiwa, Kazuhiro","last_name":"Oiwa","first_name":"Kazuhiro"},{"first_name":"Jian","full_name":"Yu, Jian","last_name":"Yu"},{"first_name":"Min","full_name":"Yao, Min","last_name":"Yao"},{"last_name":"Nakagaki","full_name":"Nakagaki, Toshiyuki","first_name":"Toshiyuki"},{"full_name":"Tamaoki, Nobuyuki","last_name":"Tamaoki","first_name":"Nobuyuki"}],"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","citation":{"ista":"Thayyil S, Nishigami Y, Islam MJ, Hashim PK, Furuta K, Oiwa K, Yu J, Yao M, Nakagaki T, Tamaoki N. 2022. Dynamic control of microbial movement by photoswitchable ATP antagonists. Chemistry - A European Journal. 28(30), e202200807.","chicago":"Thayyil, Sampreeth, Yukinori Nishigami, Muhammad J Islam, P. K. Hashim, Ken’Ya Furuta, Kazuhiro Oiwa, Jian Yu, Min Yao, Toshiyuki Nakagaki, and Nobuyuki Tamaoki. “Dynamic Control of Microbial Movement by Photoswitchable ATP Antagonists.” Chemistry - A European Journal. Wiley, 2022. https://doi.org/10.1002/chem.202200807.","apa":"Thayyil, S., Nishigami, Y., Islam, M. J., Hashim, P. K., Furuta, K., Oiwa, K., … Tamaoki, N. (2022). Dynamic control of microbial movement by photoswitchable ATP antagonists. Chemistry - A European Journal. Wiley. https://doi.org/10.1002/chem.202200807","ama":"Thayyil S, Nishigami Y, Islam MJ, et al. Dynamic control of microbial movement by photoswitchable ATP antagonists. Chemistry - A European Journal. 2022;28(30). doi:10.1002/chem.202200807","short":"S. Thayyil, Y. Nishigami, M.J. Islam, P.K. Hashim, K. Furuta, K. Oiwa, J. Yu, M. Yao, T. Nakagaki, N. Tamaoki, Chemistry - A European Journal 28 (2022).","ieee":"S. Thayyil et al., “Dynamic control of microbial movement by photoswitchable ATP antagonists,” Chemistry - A European Journal, vol. 28, no. 30. Wiley, 2022.","mla":"Thayyil, Sampreeth, et al. “Dynamic Control of Microbial Movement by Photoswitchable ATP Antagonists.” Chemistry - A European Journal, vol. 28, no. 30, e202200807, Wiley, 2022, doi:10.1002/chem.202200807."},"article_number":"e202200807","date_created":"2022-04-24T22:01:44Z","doi":"10.1002/chem.202200807","date_published":"2022-05-25T00:00:00Z","publication":"Chemistry - A European Journal","day":"25","year":"2022","isi":1,"oa":1,"publisher":"Wiley","quality_controlled":"1"},{"has_accepted_license":"1","year":"2022","day":"16","page":"108","date_published":"2022-05-16T00:00:00Z","doi":"10.15479/at:ista:11393","date_created":"2022-05-17T08:57:41Z","publisher":"Institute of Science and Technology Austria","oa":1,"citation":{"chicago":"Jevtic, Marijo. “Contextual Fear Learning Induced Changes in AMPA Receptor Subtypes along the Proximodistal Axis in Dorsal Hippocampus.” Institute of Science and Technology Austria, 2022. https://doi.org/10.15479/at:ista:11393.","ista":"Jevtic M. 2022. Contextual fear learning induced changes in AMPA receptor subtypes along the proximodistal axis in dorsal hippocampus. Institute of Science and Technology Austria.","mla":"Jevtic, Marijo. Contextual Fear Learning Induced Changes in AMPA Receptor Subtypes along the Proximodistal Axis in Dorsal Hippocampus. Institute of Science and Technology Austria, 2022, doi:10.15479/at:ista:11393.","short":"M. Jevtic, Contextual Fear Learning Induced Changes in AMPA Receptor Subtypes along the Proximodistal Axis in Dorsal Hippocampus, Institute of Science and Technology Austria, 2022.","ieee":"M. Jevtic, “Contextual fear learning induced changes in AMPA receptor subtypes along the proximodistal axis in dorsal hippocampus,” Institute of Science and Technology Austria, 2022.","ama":"Jevtic M. Contextual fear learning induced changes in AMPA receptor subtypes along the proximodistal axis in dorsal hippocampus. 2022. doi:10.15479/at:ista:11393","apa":"Jevtic, M. (2022). Contextual fear learning induced changes in AMPA receptor subtypes along the proximodistal axis in dorsal hippocampus. Institute of Science and Technology Austria. https://doi.org/10.15479/at:ista:11393"},"user_id":"8b945eb4-e2f2-11eb-945a-df72226e66a9","author":[{"first_name":"Marijo","id":"4BE3BC94-F248-11E8-B48F-1D18A9856A87","full_name":"Jevtic, Marijo","last_name":"Jevtic"}],"article_processing_charge":"No","title":"Contextual fear learning induced changes in AMPA receptor subtypes along the proximodistal axis in dorsal hippocampus","publication_identifier":{"issn":["2663-337X"]},"publication_status":"published","degree_awarded":"PhD","file":[{"access_level":"closed","relation":"source_file","content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","embargo_to":"open_access","checksum":"8fc695d88020d70d231dad0e9f10b138","file_id":"11395","creator":"cchlebak","date_updated":"2023-05-17T22:30:03Z","file_size":56427603,"date_created":"2022-05-17T09:08:06Z","file_name":"MJ thesis.docx"},{"content_type":"application/pdf","relation":"main_file","access_level":"open_access","embargo":"2023-05-16","checksum":"c1dd20a1aece521b3500607b00e463d6","file_id":"11397","file_size":4351981,"date_updated":"2023-05-17T22:30:03Z","creator":"cchlebak","file_name":"MJ_thesis_PDFA.pdf","date_created":"2022-05-17T12:09:25Z"}],"language":[{"iso":"eng"}],"related_material":{"record":[{"id":"7391","status":"public","relation":"part_of_dissertation"}]},"abstract":[{"lang":"eng","text":"AMPA receptors (AMPARs) mediate fast excitatory neurotransmission and their role is\r\nimplicated in complex processes such as learning and memory and various neurological\r\ndiseases. These receptors are composed of different subunits and the subunit composition can\r\naffect channel properties, receptor trafficking and interaction with other associated proteins.\r\nUsing the high sensitivity SDS-digested freeze-fracture replica labeling (SDS-FRL) for\r\nelectron microscopy I investigated the number, density, and localization of AMPAR subunits,\r\nGluA1, GluA2, GluA3, and GluA1-3 (panAMPA) in pyramidal cells in the CA1 area of mouse\r\nhippocampus. I have found that the immunogold labeling for all of these subunits in the\r\npostsynaptic sites was highest in stratum radiatum and lowest in stratum lacunosummoleculare. The labeling density for the all subunits in the extrasynaptic sites showed a gradual\r\nincrease from the pyramidal cell soma towards the distal part of stratum radiatum. The densities\r\nof extrasynaptic GluA1, GluA2 and panAMPA labeling reached 10-15% of synaptic densities,\r\nwhile the ratio of extrasynaptic labeling for GluA3 was significantly lower compared than those\r\nfor other subunits. The labeling patterns for GluA1, GluA2 and GluA1-3 are similar and their\r\ndensities were higher in the periphery than center of synapses. In contrast, the GluA3-\r\ncontaining receptors were more centrally localized compared to the GluA1- and GluA2-\r\ncontaining receptors.\r\nThe hippocampus plays a central role in learning and memory. Contextual learning has been\r\nshown to require the delivery of AMPA receptors to CA1 synapses in the dorsal hippocampus.\r\nHowever, proximodistal heterogeneity of this plasticity and particular contribution of different\r\nAMPA receptor subunits are not fully understood. By combining inhibitory avoidance task, a\r\nhippocampus-dependent contextual fear-learning paradigm, with SDS-FRL, I have revealed an\r\nincrease in synaptic density specific to GluA1-containing AMPA receptors in the CA1 area.\r\nThe intrasynaptic distribution of GluA1 also changed from the periphery to center-preferred\r\npattern. Furthermore, this synaptic plasticity was evident selectively in stratum radiatum but\r\nnot stratum oriens, and in the CA1 subregion proximal but not distal to CA2. These findings\r\nfurther contribute to our understanding of how specific hippocampal subregions and AMPA\r\nreceptor subunits are involved in physiological learning.\r\nAlthough the immunolabeling results above shed light on subunit-specific plasticity in\r\nAMPAR distribution, no tools to visualize and study the subunit composition at the single\r\nchannel level in situ have been available. Electron microscopy with conventional immunogold\r\nlabeling approaches has limitations in the single channel analysis because of the large size of\r\nantibodies and steric hindrance hampering multiple subunit labeling of single channels. I\r\nmanaged to develop a new chemical labeling system using a short peptide tag and small\r\nsynthetic probes, which form specific covalent bond with a cysteine residue in the tag fused to\r\nproteins of interest (reactive tag system). I additionally made substantial progress into adapting\r\nthis system for AMPA receptor subunits."}],"acknowledged_ssus":[{"_id":"EM-Fac"}],"oa_version":"Published Version","alternative_title":["ISTA Thesis"],"month":"05","supervisor":[{"last_name":"Shigemoto","orcid":"0000-0001-8761-9444","full_name":"Shigemoto, Ryuichi","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","first_name":"Ryuichi"}],"date_updated":"2023-09-07T14:53:44Z","ddc":["570"],"file_date_updated":"2023-05-17T22:30:03Z","department":[{"_id":"GradSch"},{"_id":"RySh"}],"_id":"11393","type":"dissertation","status":"public"}]