Publications by authors named "Tobias M Rasse"

24 Publications

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Autophagy inhibition rescues structural and functional defects caused by the loss of mitochondrial chaperone in .

Autophagy 2021 Jan 25:1-15. Epub 2021 Jan 25.

Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen , Tübingen, Germany.

We investigated in larval and adult models whether loss of the mitochondrial chaperone is sufficient to cause pathological alterations commonly observed in Parkinson disease. At affected larval neuromuscular junctions, no effects on terminal size, bouton size or number, synapse size, or number were observed, suggesting that we studied an early stage of pathogenesis. At this stage, we noted a loss of synaptic vesicle proteins and active zone components, delayed synapse maturation, reduced evoked and spontaneous excitatory junctional potentials, increased synaptic fatigue, and cytoskeleton rearrangements. The adult model displayed ATP depletion, altered body posture, and susceptibility to heat-induced paralysis. Adult phenotypes could be suppressed by knockdown of , and . The knockdown of components of the macroautophagy/autophagy machinery or overexpression of human broadly rescued larval and adult phenotypes, while disease-associated variants did not. Overexpression of or promotion of autophagy exacerbated defects. AEL: after egg laying; AZ: active zone; brp: bruchpilot; Csp: cysteine string protein; dlg: discs large; eEJPs: evoked excitatory junctional potentials; GluR: glutamate receptor; HO: hydrogen peroxide; mEJP: miniature excitatory junctional potentials; MT: microtubule; NMJ: neuromuscular junction; PD: Parkinson disease; : PTEN-induced putative kinase 1; PSD: postsynaptic density; SSR: subsynaptic reticulum; SV: synaptic vesicle; VGlut: vesicular glutamate transporter.
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http://dx.doi.org/10.1080/15548627.2020.1871211DOI Listing
January 2021

OpSeF: Open Source Python Framework for Collaborative Instance Segmentation of Bioimages.

Front Bioeng Biotechnol 2020 6;8:558880. Epub 2020 Oct 6.

Synthetic and Systems Biology Unit, Biological Research Center (BRC), Szeged, Hungary.

Various pre-trained deep learning models for the segmentation of bioimages have been made available as developer-to-end-user solutions. They are optimized for ease of use and usually require neither knowledge of machine learning nor coding skills. However, individually testing these tools is tedious and success is uncertain. Here, we present the Open Segmentation Framework (OpSeF), a Python framework for deep learning-based instance segmentation. OpSeF aims at facilitating the collaboration of biomedical users with experienced image analysts. It builds on the analysts' knowledge in Python, machine learning, and workflow design to solve complex analysis tasks at any scale in a reproducible, well-documented way. OpSeF defines standard inputs and outputs, thereby facilitating modular workflow design and interoperability with other software. Users play an important role in problem definition, quality control, and manual refinement of results. OpSeF semi-automates preprocessing, convolutional neural network (CNN)-based segmentation in 2D or 3D, and postprocessing. It facilitates benchmarking of multiple models in parallel. OpSeF streamlines the optimization of parameters for pre- and postprocessing such, that an available model may frequently be used without retraining. Even if sufficiently good results are not achievable with this approach, intermediate results can inform the analysts in the selection of the most promising CNN-architecture in which the biomedical user might invest the effort of manually labeling training data. We provide Jupyter notebooks that document sample workflows based on various image collections. Analysts may find these notebooks useful to illustrate common segmentation challenges, as they prepare the advanced user for gradually taking over some of their tasks and completing their projects independently. The notebooks may also be used to explore the analysis options available within OpSeF in an interactive way and to document and share final workflows. Currently, three mechanistically distinct CNN-based segmentation methods, the U-Net implementation used in Cellprofiler 3.0, StarDist, and Cellpose have been integrated within OpSeF. The addition of new networks requires little; the addition of new models requires no coding skills. Thus, OpSeF might soon become both an interactive model repository, in which pre-trained models might be shared, evaluated, and reused with ease.
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http://dx.doi.org/10.3389/fbioe.2020.558880DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7576117PMC
October 2020

Restraint of presynaptic protein levels by Wnd/DLK signaling mediates synaptic defects associated with the kinesin-3 motor Unc-104.

Elife 2017 09 19;6. Epub 2017 Sep 19.

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, United States.

The kinesin-3 family member Unc-104/KIF1A is required for axonal transport of many presynaptic components to synapses, and mutation of this gene results in synaptic dysfunction in mice, flies and worms. Our studies at the neuromuscular junction indicate that many synaptic defects in mutants are mediated independently of Unc-104's transport function, via the Wallenda (Wnd)/DLK MAP kinase axonal damage signaling pathway. Wnd signaling becomes activated when Unc-104's function is disrupted, and leads to impairment of synaptic structure and function by restraining the expression level of active zone (AZ) and synaptic vesicle (SV) components. This action concomitantly suppresses the buildup of synaptic proteins in neuronal cell bodies, hence may play an adaptive role to stresses that impair axonal transport. Wnd signaling also becomes activated when pre-synaptic proteins are over-expressed, suggesting the existence of a feedback circuit to match synaptic protein levels to the transport capacity of the axon.
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http://dx.doi.org/10.7554/eLife.24271DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5605197PMC
September 2017

The KIF1A homolog Unc-104 is important for spontaneous release, postsynaptic density maturation and perisynaptic scaffold organization.

Sci Rep 2017 03 27;7:38172. Epub 2017 Mar 27.

Junior Research Group Synaptic Plasticity, Hertie-Institute for Clinical Brain Research, University of Tübingen, Otfried-Müller-Str. 27, 72076 Tübingen 72076, Germany.

The kinesin-3 family member KIF1A has been shown to be important for experience dependent neuroplasticity. In Drosophila, amorphic mutations in the KIF1A homolog unc-104 disrupt the formation of mature boutons. Disease associated KIF1A mutations have been associated with motor and sensory dysfunctions as well as non-syndromic intellectual disability in humans. A hypomorphic mutation in the forkhead-associated domain of Unc-104, unc-104, impairs active zone maturation resulting in an increased fraction of post-synaptic glutamate receptor fields that lack the active zone scaffolding protein Bruchpilot. Here, we show that the unc-104mutation causes defects in synaptic transmission as manifested by reduced amplitude of both evoked and miniature excitatory junctional potentials. Structural defects observed in the postsynaptic compartment of mutant NMJs include reduced glutamate receptor field size, and altered glutamate receptor composition. In addition, we observed marked loss of postsynaptic scaffolding proteins and reduced complexity of the sub-synaptic reticulum, which could be rescued by pre- but not postsynaptic expression of unc-104. Our results highlight the importance of kinesin-3 based axonal transport in synaptic transmission and provide novel insights into the role of Unc-104 in synapse maturation.
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http://dx.doi.org/10.1038/srep38172DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5366810PMC
March 2017

The Drosophila KIF1A Homolog unc-104 Is Important for Site-Specific Synapse Maturation.

Front Cell Neurosci 2016 5;10:207. Epub 2016 Sep 5.

Junior Research Group Synaptic Plasticity, Hertie-Institute for Clinical Brain Research, University of TübingenTübingen, Germany; Schaller Research Group at the University of Heidelberg and DKFZ, Proteostasis in Neurodegenerative Disease (B180), German Cancer Research CenterHeidelberg, Germany.

Mutations in the kinesin-3 family member KIF1A have been associated with hereditary spastic paraplegia (HSP), hereditary and sensory autonomic neuropathy type 2 (HSAN2) and non-syndromic intellectual disability (ID). Both autosomal recessive and autosomal dominant forms of inheritance have been reported. Loss of KIF1A or its homolog unc-104 causes early postnatal or embryonic lethality in mice and Drosophila, respectively. In this study, we use a previously described hypomorphic allele of unc-104, unc-104(bris) , to investigate the impact of partial loss-of-function of kinesin-3 on synapse maturation at the Drosophila neuromuscular junction (NMJ). Unc-104(bris) mutants exhibit structural defects where a subset of synapses at the NMJ lack all investigated active zone (AZ) proteins, suggesting a complete failure in the formation of the cytomatrix at the active zone (CAZ) at these sites. Modulating synaptic Bruchpilot (Brp) levels by ectopic overexpression or RNAi-mediated knockdown suggests that the loss of AZ components such as Ca(2+) channels and Liprin-α is caused by impaired kinesin-3 based transport rather than due to the absence of the key AZ organizer protein, Brp. In addition to defects in CAZ assembly, unc-104(bris) mutants display further defects such as depletion of dense core and synaptic vesicle (SV) markers from the NMJ. Notably, the level of Rab3, which is important for the allocation of AZ proteins to individual release sites, was severely reduced at unc-104(bris) mutant NMJs. Overexpression of Rab3 partially ameliorates synaptic phenotypes of unc-104(bris) larvae, suggesting that lack of presynaptic Rab3 contributes to defects in synapse maturation.
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http://dx.doi.org/10.3389/fncel.2016.00207DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5011141PMC
September 2016

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).

Authors:
Daniel J Klionsky Kotb Abdelmohsen Akihisa Abe Md Joynal Abedin Hagai Abeliovich Abraham Acevedo Arozena Hiroaki Adachi Christopher M Adams Peter D Adams Khosrow Adeli Peter J Adhihetty Sharon G Adler Galila Agam Rajesh Agarwal Manish K Aghi Maria Agnello Patrizia Agostinis Patricia V Aguilar Julio Aguirre-Ghiso Edoardo M Airoldi Slimane Ait-Si-Ali Takahiko Akematsu Emmanuel T Akporiaye Mohamed Al-Rubeai Guillermo M Albaiceta Chris Albanese Diego Albani Matthew L Albert Jesus Aldudo Hana Algül Mehrdad Alirezaei Iraide Alloza Alexandru Almasan Maylin Almonte-Beceril Emad S Alnemri Covadonga Alonso Nihal Altan-Bonnet Dario C Altieri Silvia Alvarez Lydia Alvarez-Erviti Sandro Alves Giuseppina Amadoro Atsuo Amano Consuelo Amantini Santiago Ambrosio Ivano Amelio Amal O Amer Mohamed Amessou Angelika Amon Zhenyi An Frank A Anania Stig U Andersen Usha P Andley Catherine K Andreadi Nathalie Andrieu-Abadie Alberto Anel David K Ann Shailendra Anoopkumar-Dukie Manuela Antonioli Hiroshi Aoki Nadezda Apostolova Saveria Aquila Katia Aquilano Koichi Araki Eli Arama Agustin Aranda Jun Araya Alexandre Arcaro Esperanza Arias Hirokazu Arimoto Aileen R Ariosa Jane L Armstrong Thierry Arnould Ivica Arsov Katsuhiko Asanuma Valerie Askanas Eric Asselin Ryuichiro Atarashi Sally S Atherton Julie D Atkin Laura D Attardi Patrick Auberger Georg Auburger Laure Aurelian Riccardo Autelli Laura Avagliano Maria Laura Avantaggiati Limor Avrahami Suresh Awale Neelam Azad Tiziana Bachetti Jonathan M Backer Dong-Hun Bae Jae-Sung Bae Ok-Nam Bae Soo Han Bae Eric H Baehrecke Seung-Hoon Baek Stephen Baghdiguian Agnieszka Bagniewska-Zadworna Hua Bai Jie Bai Xue-Yuan Bai Yannick Bailly Kithiganahalli Narayanaswamy Balaji Walter Balduini Andrea Ballabio Rena Balzan Rajkumar Banerjee Gábor Bánhegyi Haijun Bao Benoit Barbeau Maria D Barrachina Esther Barreiro Bonnie Bartel Alberto Bartolomé Diane C Bassham Maria Teresa Bassi Robert C Bast Alakananda Basu Maria Teresa Batista Henri Batoko Maurizio Battino Kyle Bauckman Bradley L Baumgarner K Ulrich Bayer Rupert Beale Jean-François Beaulieu George R Beck Christoph Becker J David Beckham Pierre-André Bédard Patrick J Bednarski Thomas J Begley Christian Behl Christian Behrends Georg Mn Behrens Kevin E Behrns Eloy Bejarano Amine Belaid Francesca Belleudi Giovanni Bénard Guy Berchem Daniele Bergamaschi Matteo Bergami Ben Berkhout Laura Berliocchi Amélie Bernard Monique Bernard Francesca Bernassola Anne Bertolotti Amanda S Bess Sébastien Besteiro Saverio Bettuzzi Savita Bhalla Shalmoli Bhattacharyya Sujit K Bhutia Caroline Biagosch Michele Wolfe Bianchi Martine Biard-Piechaczyk Viktor Billes Claudia Bincoletto Baris Bingol Sara W Bird Marc Bitoun Ivana Bjedov Craig Blackstone Lionel Blanc Guillermo A Blanco Heidi Kiil Blomhoff Emilio Boada-Romero Stefan Böckler Marianne Boes Kathleen Boesze-Battaglia Lawrence H Boise Alessandra Bolino Andrea Boman Paolo Bonaldo Matteo Bordi Jürgen Bosch Luis M Botana Joelle Botti German Bou Marina Bouché Marion Bouchecareilh Marie-Josée Boucher Michael E Boulton Sebastien G Bouret Patricia Boya Michaël Boyer-Guittaut Peter V Bozhkov Nathan Brady Vania Mm Braga Claudio Brancolini Gerhard H Braus José M Bravo-San Pedro Lisa A Brennan Emery H Bresnick Patrick Brest Dave Bridges Marie-Agnès Bringer Marisa Brini Glauber C Brito Bertha Brodin Paul S Brookes Eric J Brown Karen Brown Hal E Broxmeyer Alain Bruhat Patricia Chakur Brum John H Brumell Nicola Brunetti-Pierri Robert J Bryson-Richardson Shilpa Buch Alastair M Buchan Hikmet Budak Dmitry V Bulavin Scott J Bultman Geert Bultynck Vladimir Bumbasirevic Yan Burelle Robert E Burke Margit Burmeister Peter Bütikofer Laura Caberlotto Ken Cadwell Monika Cahova Dongsheng Cai Jingjing Cai Qian Cai Sara Calatayud Nadine Camougrand Michelangelo Campanella Grant R Campbell Matthew Campbell Silvia Campello Robin Candau Isabella Caniggia Lavinia Cantoni Lizhi Cao Allan B Caplan Michele Caraglia Claudio Cardinali Sandra Morais Cardoso Jennifer S Carew Laura A Carleton Cathleen R Carlin Silvia Carloni Sven R Carlsson Didac Carmona-Gutierrez Leticia Am Carneiro Oliana Carnevali Serena Carra Alice Carrier Bernadette Carroll Caty Casas Josefina Casas Giuliana Cassinelli Perrine Castets Susana Castro-Obregon Gabriella Cavallini Isabella Ceccherini Francesco Cecconi Arthur I Cederbaum Valentín Ceña Simone Cenci Claudia Cerella Davide Cervia Silvia Cetrullo Hassan Chaachouay Han-Jung Chae Andrei S Chagin Chee-Yin Chai Gopal Chakrabarti Georgios Chamilos Edmond Yw Chan Matthew Tv Chan Dhyan Chandra Pallavi Chandra Chih-Peng Chang Raymond Chuen-Chung Chang Ta Yuan Chang John C Chatham Saurabh Chatterjee Santosh Chauhan Yongsheng Che Michael E Cheetham Rajkumar Cheluvappa Chun-Jung Chen Gang Chen Guang-Chao Chen Guoqiang Chen Hongzhuan Chen Jeff W Chen Jian-Kang Chen Min Chen Mingzhou Chen Peiwen Chen Qi Chen Quan Chen Shang-Der Chen Si Chen Steve S-L Chen Wei Chen Wei-Jung Chen Wen Qiang Chen Wenli Chen Xiangmei Chen Yau-Hung Chen Ye-Guang Chen Yin Chen Yingyu Chen Yongshun Chen Yu-Jen Chen Yue-Qin Chen Yujie Chen Zhen Chen Zhong Chen Alan Cheng Christopher Hk Cheng Hua Cheng Heesun Cheong Sara Cherry Jason Chesney Chun Hei Antonio Cheung Eric Chevet Hsiang Cheng Chi Sung-Gil Chi Fulvio Chiacchiera Hui-Ling Chiang Roberto Chiarelli Mario Chiariello Marcello Chieppa Lih-Shen Chin Mario Chiong Gigi Nc Chiu Dong-Hyung Cho Ssang-Goo Cho William C Cho Yong-Yeon Cho Young-Seok Cho Augustine Mk Choi Eui-Ju Choi Eun-Kyoung Choi Jayoung Choi Mary E Choi Seung-Il Choi Tsui-Fen Chou Salem Chouaib Divaker Choubey Vinay Choubey Kuan-Chih Chow Kamal Chowdhury Charleen T Chu Tsung-Hsien Chuang Taehoon Chun Hyewon Chung Taijoon Chung Yuen-Li Chung Yong-Joon Chwae Valentina Cianfanelli Roberto Ciarcia Iwona A Ciechomska Maria Rosa Ciriolo Mara Cirone Sofie Claerhout Michael J Clague Joan Clària Peter Gh Clarke Robert Clarke Emilio Clementi Cédric Cleyrat Miriam Cnop Eliana M Coccia Tiziana Cocco Patrice Codogno Jörn Coers Ezra Ew Cohen David Colecchia Luisa Coletto Núria S Coll Emma Colucci-Guyon Sergio Comincini Maria Condello Katherine L Cook Graham H Coombs Cynthia D Cooper J Mark Cooper Isabelle Coppens Maria Tiziana Corasaniti Marco Corazzari Ramon Corbalan Elisabeth Corcelle-Termeau Mario D Cordero Cristina Corral-Ramos Olga Corti Andrea Cossarizza Paola Costelli Safia Costes Susan L Cotman Ana Coto-Montes Sandra Cottet Eduardo Couve Lori R Covey L Ashley Cowart Jeffery S Cox Fraser P Coxon Carolyn B Coyne Mark S Cragg Rolf J Craven Tiziana Crepaldi Jose L Crespo Alfredo Criollo Valeria Crippa Maria Teresa Cruz Ana Maria Cuervo Jose M Cuezva Taixing Cui Pedro R Cutillas Mark J Czaja Maria F Czyzyk-Krzeska Ruben K Dagda Uta Dahmen Chunsun Dai Wenjie Dai Yun Dai Kevin N Dalby Luisa Dalla Valle Guillaume Dalmasso Marcello D'Amelio Markus Damme Arlette Darfeuille-Michaud Catherine Dargemont Victor M Darley-Usmar Srinivasan Dasarathy Biplab Dasgupta Srikanta Dash Crispin R Dass Hazel Marie Davey Lester M Davids David Dávila Roger J Davis Ted M Dawson Valina L Dawson Paula Daza Jackie de Belleroche Paul de Figueiredo Regina Celia Bressan Queiroz de Figueiredo José de la Fuente Luisa De Martino Antonella De Matteis Guido Ry De Meyer Angelo De Milito Mauro De Santi Wanderley de Souza Vincenzo De Tata Daniela De Zio Jayanta Debnath Reinhard Dechant Jean-Paul Decuypere Shane Deegan Benjamin Dehay Barbara Del Bello Dominic P Del Re Régis Delage-Mourroux Lea Md Delbridge Louise Deldicque Elizabeth Delorme-Axford Yizhen Deng Joern Dengjel Melanie Denizot Paul Dent Channing J Der Vojo Deretic Benoît Derrien Eric Deutsch Timothy P Devarenne Rodney J Devenish Sabrina Di Bartolomeo Nicola Di Daniele Fabio Di Domenico Alessia Di Nardo Simone Di Paola Antonio Di Pietro Livia Di Renzo Aaron DiAntonio Guillermo Díaz-Araya Ines Díaz-Laviada Maria T Diaz-Meco Javier Diaz-Nido Chad A Dickey Robert C Dickson Marc Diederich Paul Digard Ivan Dikic Savithrama P Dinesh-Kumar Chan Ding Wen-Xing Ding Zufeng Ding Luciana Dini Jörg Hw Distler Abhinav Diwan Mojgan Djavaheri-Mergny Kostyantyn Dmytruk Renwick Cj Dobson Volker Doetsch Karol Dokladny Svetlana Dokudovskaya Massimo Donadelli X Charlie Dong Xiaonan Dong Zheng Dong Terrence M Donohue Kelly S Doran Gabriella D'Orazi Gerald W Dorn Victor Dosenko Sami Dridi Liat Drucker Jie Du Li-Lin Du Lihuan Du André du Toit Priyamvada Dua Lei Duan Pu Duann Vikash Kumar Dubey Michael R Duchen Michel A Duchosal Helene Duez Isabelle Dugail Verónica I Dumit Mara C Duncan Elaine A Dunlop William A Dunn Nicolas Dupont Luc Dupuis Raúl V Durán Thomas M Durcan Stéphane Duvezin-Caubet Umamaheswar Duvvuri Vinay Eapen Darius Ebrahimi-Fakhari Arnaud Echard Leopold Eckhart Charles L Edelstein Aimee L Edinger Ludwig Eichinger Tobias Eisenberg Avital Eisenberg-Lerner N Tony Eissa Wafik S El-Deiry Victoria El-Khoury Zvulun Elazar Hagit Eldar-Finkelman Chris Jh Elliott Enzo Emanuele Urban Emmenegger Nikolai Engedal Anna-Mart Engelbrecht Simone Engelender Jorrit M Enserink Ralf Erdmann Jekaterina Erenpreisa Rajaraman Eri Jason L Eriksen Andreja Erman Ricardo Escalante Eeva-Liisa Eskelinen Lucile Espert Lorena Esteban-Martínez Thomas J Evans Mario Fabri Gemma Fabrias Cinzia Fabrizi Antonio Facchiano Nils J Færgeman Alberto Faggioni W Douglas Fairlie Chunhai Fan Daping Fan Jie Fan Shengyun Fang Manolis Fanto Alessandro Fanzani Thomas Farkas Mathias Faure Francois B Favier Howard Fearnhead Massimo Federici Erkang Fei Tania C Felizardo Hua Feng Yibin Feng Yuchen Feng Thomas A Ferguson Álvaro F Fernández Maite G Fernandez-Barrena Jose C Fernandez-Checa Arsenio Fernández-López Martin E Fernandez-Zapico Olivier Feron Elisabetta Ferraro Carmen Veríssima Ferreira-Halder Laszlo Fesus Ralph Feuer Fabienne C Fiesel Eduardo C Filippi-Chiela Giuseppe Filomeni Gian Maria Fimia John H Fingert Steven Finkbeiner Toren Finkel Filomena Fiorito Paul B Fisher Marc Flajolet Flavio Flamigni Oliver Florey Salvatore Florio R Andres Floto Marco Folini Carlo Follo Edward A Fon Francesco Fornai Franco Fortunato Alessandro Fraldi Rodrigo Franco Arnaud Francois Aurélie François Lisa B Frankel Iain Dc Fraser Norbert Frey Damien G Freyssenet Christian Frezza Scott L Friedman Daniel E Frigo Dongxu Fu José M Fuentes Juan Fueyo Yoshio Fujitani Yuuki Fujiwara Mikihiro Fujiya Mitsunori Fukuda Simone Fulda Carmela Fusco Bozena Gabryel Matthias Gaestel Philippe Gailly Malgorzata Gajewska Sehamuddin Galadari Gad Galili Inmaculada Galindo Maria F Galindo Giovanna Galliciotti Lorenzo Galluzzi Luca Galluzzi Vincent Galy Noor Gammoh Sam Gandy Anand K Ganesan Swamynathan Ganesan Ian G Ganley Monique Gannagé Fen-Biao Gao Feng Gao Jian-Xin Gao Lorena García Nannig Eleonora García Véscovi Marina Garcia-Macía Carmen Garcia-Ruiz Abhishek D Garg Pramod Kumar Garg Ricardo Gargini Nils Christian Gassen Damián Gatica Evelina Gatti Julie Gavard Evripidis Gavathiotis Liang Ge Pengfei Ge Shengfang Ge Po-Wu Gean Vania Gelmetti Armando A Genazzani Jiefei Geng Pascal Genschik Lisa Gerner Jason E Gestwicki David A Gewirtz Saeid Ghavami Eric Ghigo Debabrata Ghosh Anna Maria Giammarioli Francesca Giampieri Claudia Giampietri Alexandra Giatromanolaki Derrick J Gibbings Lara Gibellini Spencer B Gibson Vanessa Ginet Antonio Giordano Flaviano Giorgini Elisa Giovannetti Stephen E Girardin Suzana Gispert Sandy Giuliano Candece L Gladson Alvaro Glavic Martin Gleave Nelly Godefroy Robert M Gogal Kuppan Gokulan Gustavo H Goldman Delia Goletti Michael S Goligorsky Aldrin V Gomes Ligia C Gomes Hernando Gomez Candelaria Gomez-Manzano Rubén Gómez-Sánchez Dawit Ap Gonçalves Ebru Goncu Qingqiu Gong Céline Gongora Carlos B Gonzalez Pedro Gonzalez-Alegre Pilar Gonzalez-Cabo Rosa Ana González-Polo Ing Swie Goping Carlos Gorbea Nikolai V Gorbunov Daphne R Goring Adrienne M Gorman Sharon M Gorski Sandro Goruppi Shino Goto-Yamada Cecilia Gotor Roberta A Gottlieb Illana Gozes Devrim Gozuacik Yacine Graba Martin Graef Giovanna E Granato Gary Dean Grant Steven Grant Giovanni Luca Gravina Douglas R Green Alexander Greenhough Michael T Greenwood Benedetto Grimaldi Frédéric Gros Charles Grose Jean-Francois Groulx Florian Gruber Paolo Grumati Tilman Grune Jun-Lin Guan Kun-Liang Guan Barbara Guerra Carlos Guillen Kailash Gulshan Jan Gunst Chuanyong Guo Lei Guo Ming Guo Wenjie Guo Xu-Guang Guo Andrea A Gust Åsa B Gustafsson Elaine Gutierrez Maximiliano G Gutierrez Ho-Shin Gwak Albert Haas James E Haber Shinji Hadano Monica Hagedorn David R Hahn Andrew J Halayko Anne Hamacher-Brady Kozo Hamada Ahmed Hamai Andrea Hamann Maho Hamasaki Isabelle Hamer Qutayba Hamid Ester M Hammond Feng Han Weidong Han James T Handa John A Hanover Malene Hansen Masaru Harada Ljubica Harhaji-Trajkovic J Wade Harper Abdel Halim Harrath Adrian L Harris James Harris Udo Hasler Peter Hasselblatt Kazuhisa Hasui Robert G Hawley Teresa S Hawley Congcong He Cynthia Y He Fengtian He Gu He Rong-Rong He Xian-Hui He You-Wen He Yu-Ying He Joan K Heath Marie-Josée Hébert Robert A Heinzen Gudmundur Vignir Helgason Michael Hensel Elizabeth P Henske Chengtao Her Paul K Herman Agustín Hernández Carlos Hernandez Sonia Hernández-Tiedra Claudio Hetz P Robin Hiesinger Katsumi Higaki Sabine Hilfiker Bradford G Hill Joseph A Hill William D Hill Keisuke Hino Daniel Hofius Paul Hofman Günter U Höglinger Jörg Höhfeld Marina K Holz Yonggeun Hong David A Hood Jeroen Jm Hoozemans Thorsten Hoppe Chin Hsu Chin-Yuan Hsu Li-Chung Hsu Dong Hu Guochang Hu Hong-Ming Hu Hongbo Hu Ming Chang Hu Yu-Chen Hu Zhuo-Wei Hu Fang Hua Ya Hua Canhua Huang Huey-Lan Huang Kuo-How Huang Kuo-Yang Huang Shile Huang Shiqian Huang Wei-Pang Huang Yi-Ran Huang Yong Huang Yunfei Huang Tobias B Huber Patricia Huebbe Won-Ki Huh Juha J Hulmi Gang Min Hur James H Hurley Zvenyslava Husak Sabah Na Hussain Salik Hussain Jung Jin Hwang Seungmin Hwang Thomas Is Hwang Atsuhiro Ichihara Yuzuru Imai Carol Imbriano Megumi Inomata Takeshi Into Valentina Iovane Juan L Iovanna Renato V Iozzo Nancy Y Ip Javier E Irazoqui Pablo Iribarren Yoshitaka Isaka Aleksandra J Isakovic Harry Ischiropoulos Jeffrey S Isenberg Mohammad Ishaq Hiroyuki Ishida Isao Ishii Jane E Ishmael Ciro Isidoro Ken-Ichi Isobe Erika Isono Shohreh Issazadeh-Navikas Koji Itahana Eisuke Itakura Andrei I Ivanov Anand Krishnan V Iyer José M Izquierdo Yotaro Izumi Valentina Izzo Marja Jäättelä Nadia Jaber Daniel John Jackson William T Jackson Tony George Jacob Thomas S Jacques Chinnaswamy Jagannath Ashish Jain Nihar Ranjan Jana Byoung Kuk Jang Alkesh Jani Bassam Janji Paulo Roberto Jannig Patric J Jansson Steve Jean Marina Jendrach Ju-Hong Jeon Niels Jessen Eui-Bae Jeung Kailiang Jia Lijun Jia Hong Jiang Hongchi Jiang Liwen Jiang Teng Jiang Xiaoyan Jiang Xuejun Jiang Xuejun Jiang Ying Jiang Yongjun Jiang Alberto Jiménez Cheng Jin Hongchuan Jin Lei Jin Meiyan Jin Shengkan Jin Umesh Kumar Jinwal Eun-Kyeong Jo Terje Johansen Daniel E Johnson Gail Vw Johnson James D Johnson Eric Jonasch Chris Jones Leo Ab Joosten Joaquin Jordan Anna-Maria Joseph Bertrand Joseph Annie M Joubert Dianwen Ju Jingfang Ju Hsueh-Fen Juan Katrin Juenemann Gábor Juhász Hye Seung Jung Jae U Jung Yong-Keun Jung Heinz Jungbluth Matthew J Justice Barry Jutten Nadeem O Kaakoush Kai Kaarniranta Allen Kaasik Tomohiro Kabuta Bertrand Kaeffer Katarina Kågedal Alon Kahana Shingo Kajimura Or Kakhlon Manjula Kalia Dhan V Kalvakolanu Yoshiaki Kamada Konstantinos Kambas Vitaliy O Kaminskyy Harm H Kampinga Mustapha Kandouz Chanhee Kang Rui Kang Tae-Cheon Kang Tomotake Kanki Thirumala-Devi Kanneganti Haruo Kanno Anumantha G Kanthasamy Marc Kantorow Maria Kaparakis-Liaskos Orsolya Kapuy Vassiliki Karantza Md Razaul Karim Parimal Karmakar Arthur Kaser Susmita Kaushik Thomas Kawula A Murat Kaynar Po-Yuan Ke Zun-Ji Ke John H Kehrl Kate E Keller Jongsook Kim Kemper Anne K Kenworthy Oliver Kepp Andreas Kern Santosh Kesari David Kessel Robin Ketteler Isis do Carmo Kettelhut Bilon Khambu Muzamil Majid Khan Vinoth Km Khandelwal Sangeeta Khare Juliann G Kiang Amy A Kiger Akio Kihara Arianna L Kim Cheol Hyeon Kim Deok Ryong Kim Do-Hyung Kim Eung Kweon Kim Hye Young Kim Hyung-Ryong Kim Jae-Sung Kim Jeong Hun Kim Jin Cheon Kim Jin Hyoung Kim Kwang Woon Kim Michael D Kim Moon-Moo Kim Peter K Kim Seong Who Kim Soo-Youl Kim Yong-Sun Kim Yonghyun Kim Adi Kimchi Alec C Kimmelman Tomonori Kimura Jason S King Karla Kirkegaard Vladimir Kirkin Lorrie A Kirshenbaum Shuji Kishi Yasuo Kitajima Katsuhiko Kitamoto Yasushi Kitaoka Kaio Kitazato Rudolf A Kley Walter T Klimecki Michael Klinkenberg Jochen Klucken Helene Knævelsrud Erwin Knecht Laura Knuppertz Jiunn-Liang Ko Satoru Kobayashi Jan C Koch Christelle Koechlin-Ramonatxo Ulrich Koenig Young Ho Koh Katja Köhler Sepp D Kohlwein Masato Koike Masaaki Komatsu Eiki Kominami Dexin Kong Hee Jeong Kong Eumorphia G Konstantakou Benjamin T Kopp Tamas Korcsmaros Laura Korhonen Viktor I Korolchuk Nadya V Koshkina Yanjun Kou Michael I Koukourakis Constantinos Koumenis Attila L Kovács Tibor Kovács Werner J Kovacs Daisuke Koya Claudine Kraft Dimitri Krainc Helmut Kramer Tamara Kravic-Stevovic Wilhelm Krek Carole Kretz-Remy Roswitha Krick Malathi Krishnamurthy Janos Kriston-Vizi Guido Kroemer Michael C Kruer Rejko Kruger Nicholas T Ktistakis Kazuyuki Kuchitsu Christian Kuhn Addanki Pratap Kumar Anuj Kumar Ashok Kumar Deepak Kumar Dhiraj Kumar Rakesh Kumar Sharad Kumar Mondira Kundu Hsing-Jien Kung Atsushi Kuno Sheng-Han Kuo Jeff Kuret Tino Kurz Terry Kwok Taeg Kyu Kwon Yong Tae Kwon Irene Kyrmizi Albert R La Spada Frank Lafont Tim Lahm Aparna Lakkaraju Truong Lam Trond Lamark Steve Lancel Terry H Landowski Darius J R Lane Jon D Lane Cinzia Lanzi Pierre Lapaquette Louis R Lapierre Jocelyn Laporte Johanna Laukkarinen Gordon W Laurie Sergio Lavandero Lena Lavie Matthew J LaVoie Betty Yuen Kwan Law Helen Ka-Wai Law Kelsey B Law Robert Layfield Pedro A Lazo Laurent Le Cam Karine G Le Roch Hervé Le Stunff Vijittra Leardkamolkarn Marc Lecuit Byung-Hoon Lee Che-Hsin Lee Erinna F Lee Gyun Min Lee He-Jin Lee Hsinyu Lee Jae Keun Lee Jongdae Lee Ju-Hyun Lee Jun Hee Lee Michael Lee Myung-Shik Lee Patty J Lee Sam W Lee Seung-Jae Lee Shiow-Ju Lee Stella Y Lee Sug Hyung Lee Sung Sik Lee Sung-Joon Lee Sunhee Lee Ying-Ray Lee Yong J Lee Young H Lee Christiaan Leeuwenburgh Sylvain Lefort Renaud Legouis Jinzhi Lei Qun-Ying Lei David A Leib Gil Leibowitz Istvan Lekli Stéphane D Lemaire John J Lemasters Marius K Lemberg Antoinette Lemoine Shuilong Leng Guido Lenz Paola Lenzi Lilach O Lerman Daniele Lettieri Barbato Julia I-Ju Leu Hing Y Leung Beth Levine Patrick A Lewis Frank Lezoualc'h Chi Li Faqiang Li Feng-Jun Li Jun Li Ke Li Lian Li Min Li Min Li Qiang Li Rui Li Sheng Li Wei Li Wei Li Xiaotao Li Yumin Li Jiqin Lian Chengyu Liang Qiangrong Liang Yulin Liao Joana Liberal Pawel P Liberski Pearl Lie Andrew P Lieberman Hyunjung Jade Lim Kah-Leong Lim Kyu Lim Raquel T Lima Chang-Shen Lin Chiou-Feng Lin Fang Lin Fangming Lin Fu-Cheng Lin Kui Lin Kwang-Huei Lin Pei-Hui Lin Tianwei Lin Wan-Wan Lin Yee-Shin Lin Yong Lin Rafael Linden Dan Lindholm Lisa M Lindqvist Paul Lingor Andreas Linkermann Lance A Liotta Marta M Lipinski Vitor A Lira Michael P Lisanti Paloma B Liton Bo Liu Chong Liu Chun-Feng Liu Fei Liu Hung-Jen Liu Jianxun Liu Jing-Jing Liu Jing-Lan Liu Ke Liu Leyuan Liu Liang Liu Quentin Liu Rong-Yu Liu Shiming Liu Shuwen Liu Wei Liu Xian-De Liu Xiangguo Liu Xiao-Hong Liu Xinfeng Liu Xu Liu Xueqin Liu Yang Liu Yule Liu Zexian Liu Zhe Liu Juan P Liuzzi Gérard Lizard Mila Ljujic Irfan J Lodhi Susan E Logue Bal L Lokeshwar Yun Chau Long Sagar Lonial Benjamin Loos Carlos López-Otín Cristina López-Vicario Mar Lorente Philip L Lorenzi Péter Lõrincz Marek Los Michael T Lotze Penny E Lovat Binfeng Lu Bo Lu Jiahong Lu Qing Lu She-Min Lu Shuyan Lu Yingying Lu Frédéric Luciano Shirley Luckhart John Milton Lucocq Paula Ludovico Aurelia Lugea Nicholas W Lukacs Julian J Lum Anders H Lund Honglin Luo Jia Luo Shouqing Luo Claudio Luparello Timothy Lyons Jianjie Ma Yi Ma Yong Ma Zhenyi Ma Juliano Machado Glaucia M Machado-Santelli Fernando Macian Gustavo C MacIntosh Jeffrey P MacKeigan Kay F Macleod John D MacMicking Lee Ann MacMillan-Crow Frank Madeo Muniswamy Madesh Julio Madrigal-Matute Akiko Maeda Tatsuya Maeda Gustavo Maegawa Emilia Maellaro Hannelore Maes Marta Magariños Kenneth Maiese Tapas K Maiti Luigi Maiuri Maria Chiara Maiuri Carl G Maki Roland Malli Walter Malorni Alina Maloyan Fathia Mami-Chouaib Na Man Joseph D Mancias Eva-Maria Mandelkow Michael A Mandell Angelo A Manfredi Serge N Manié Claudia Manzoni Kai Mao Zixu Mao Zong-Wan Mao Philippe Marambaud Anna Maria Marconi Zvonimir Marelja Gabriella Marfe Marta Margeta Eva Margittai Muriel Mari Francesca V Mariani Concepcio Marin Sara Marinelli Guillermo Mariño Ivanka Markovic Rebecca Marquez Alberto M Martelli Sascha Martens Katie R Martin Seamus J Martin Shaun Martin Miguel A Martin-Acebes Paloma Martín-Sanz Camille Martinand-Mari Wim Martinet Jennifer Martinez Nuria Martinez-Lopez Ubaldo Martinez-Outschoorn Moisés Martínez-Velázquez Marta Martinez-Vicente Waleska Kerllen Martins Hirosato Mashima James A Mastrianni Giuseppe Matarese Paola Matarrese Roberto Mateo Satoaki Matoba Naomichi Matsumoto Takehiko Matsushita Akira Matsuura Takeshi Matsuzawa Mark P Mattson Soledad Matus Norma Maugeri Caroline Mauvezin Andreas Mayer Dusica Maysinger Guillermo D Mazzolini Mary Kate McBrayer Kimberly McCall Craig McCormick Gerald M McInerney Skye C McIver Sharon McKenna John J McMahon Iain A McNeish Fatima Mechta-Grigoriou Jan Paul Medema Diego L Medina Klara Megyeri Maryam Mehrpour Jawahar L Mehta Yide Mei Ute-Christiane Meier Alfred J Meijer Alicia Meléndez Gerry Melino Sonia Melino Edesio Jose Tenorio de Melo Maria A Mena Marc D Meneghini Javier A Menendez Regina Menezes Liesu Meng Ling-Hua Meng Songshu Meng Rossella Menghini A Sue Menko Rubem Fs Menna-Barreto Manoj B Menon Marco A Meraz-Ríos Giuseppe Merla Luciano Merlini Angelica M Merlot Andreas Meryk Stefania Meschini Joel N Meyer Man-Tian Mi Chao-Yu Miao Lucia Micale Simon Michaeli Carine Michiels Anna Rita Migliaccio Anastasia Susie Mihailidou Dalibor Mijaljica Katsuhiko Mikoshiba Enrico Milan Leonor Miller-Fleming Gordon B Mills Ian G Mills Georgia Minakaki Berge A Minassian Xiu-Fen Ming Farida Minibayeva Elena A Minina Justine D Mintern Saverio Minucci Antonio Miranda-Vizuete Claire H Mitchell Shigeki Miyamoto Keisuke Miyazawa Noboru Mizushima Katarzyna Mnich Baharia Mograbi Simin Mohseni Luis Ferreira Moita Marco Molinari Maurizio Molinari Andreas Buch Møller Bertrand Mollereau Faustino Mollinedo Marco Mongillo Martha M Monick Serena Montagnaro Craig Montell Darren J Moore Michael N Moore Rodrigo Mora-Rodriguez Paula I Moreira Etienne Morel Maria Beatrice Morelli Sandra Moreno Michael J Morgan Arnaud Moris Yuji Moriyasu Janna L Morrison Lynda A Morrison Eugenia Morselli Jorge Moscat Pope L Moseley Serge Mostowy Elisa Motori Denis Mottet Jeremy C Mottram Charbel E-H Moussa Vassiliki E Mpakou Hasan Mukhtar Jean M Mulcahy Levy Sylviane Muller Raquel Muñoz-Moreno Cristina Muñoz-Pinedo Christian Münz Maureen E Murphy James T Murray Aditya Murthy Indira U Mysorekar Ivan R Nabi Massimo Nabissi Gustavo A Nader Yukitoshi Nagahara Yoshitaka Nagai Kazuhiro Nagata Anika Nagelkerke Péter Nagy Samisubbu R Naidu Sreejayan Nair Hiroyasu Nakano Hitoshi Nakatogawa Meera Nanjundan Gennaro Napolitano Naweed I Naqvi Roberta Nardacci Derek P Narendra Masashi Narita Anna Chiara Nascimbeni Ramesh Natarajan Luiz C Navegantes Steffan T Nawrocki Taras Y Nazarko Volodymyr Y Nazarko Thomas Neill Luca M Neri Mihai G Netea Romana T Netea-Maier Bruno M Neves Paul A Ney Ioannis P Nezis Hang Tt Nguyen Huu Phuc Nguyen Anne-Sophie Nicot Hilde Nilsen Per Nilsson Mikio Nishimura Ichizo Nishino Mireia Niso-Santano Hua Niu Ralph A Nixon Vincent Co Njar Takeshi Noda Angelika A Noegel Elsie Magdalena Nolte Erik Norberg Koenraad K Norga Sakineh Kazemi Noureini Shoji Notomi Lucia Notterpek Karin Nowikovsky Nobuyuki Nukina Thorsten Nürnberger Valerie B O'Donnell Tracey O'Donovan Peter J O'Dwyer Ina Oehme Clara L Oeste Michinaga Ogawa Besim Ogretmen Yuji Ogura Young J Oh Masaki Ohmuraya Takayuki Ohshima Rani Ojha Koji Okamoto Toshiro Okazaki F Javier Oliver Karin Ollinger Stefan Olsson Daniel P Orban Paulina Ordonez Idil Orhon Laszlo Orosz Eyleen J O'Rourke Helena Orozco Angel L Ortega Elena Ortona Laura D Osellame Junko Oshima Shigeru Oshima Heinz D Osiewacz Takanobu Otomo Kinya Otsu Jing-Hsiung James Ou Tiago F Outeiro Dong-Yun Ouyang Hongjiao Ouyang Michael Overholtzer Michelle A Ozbun P Hande Ozdinler Bulent Ozpolat Consiglia Pacelli Paolo Paganetti Guylène Page Gilles Pages Ugo Pagnini Beata Pajak Stephen C Pak Karolina Pakos-Zebrucka Nazzy Pakpour Zdena Palková Francesca Palladino Kathrin Pallauf Nicolas Pallet Marta Palmieri Søren R Paludan Camilla Palumbo Silvia Palumbo Olatz Pampliega Hongming Pan Wei Pan Theocharis Panaretakis Aseem Pandey Areti Pantazopoulou Zuzana Papackova Daniela L Papademetrio Issidora Papassideri Alessio Papini Nirmala Parajuli Julian Pardo Vrajesh V Parekh Giancarlo Parenti Jong-In Park Junsoo Park Ohkmae K Park Roy Parker Rosanna Parlato Jan B Parys Katherine R Parzych Jean-Max Pasquet Benoit Pasquier Kishore Bs Pasumarthi Daniel Patschan Cam Patterson Sophie Pattingre Scott Pattison Arnim Pause Hermann Pavenstädt Flaminia Pavone Zully Pedrozo Fernando J Peña Miguel A Peñalva Mario Pende Jianxin Peng Fabio Penna Josef M Penninger Anna Pensalfini Salvatore Pepe Gustavo Js Pereira Paulo C Pereira Verónica Pérez-de la Cruz María Esther Pérez-Pérez Diego Pérez-Rodríguez Dolores Pérez-Sala Celine Perier Andras Perl David H Perlmutter Ida Perrotta Shazib Pervaiz Maija Pesonen Jeffrey E Pessin Godefridus J Peters Morten Petersen Irina Petrache Basil J Petrof Goran Petrovski James M Phang Mauro Piacentini Marina Pierdominici Philippe Pierre Valérie Pierrefite-Carle Federico Pietrocola Felipe X Pimentel-Muiños Mario Pinar Benjamin Pineda Ronit Pinkas-Kramarski Marcello Pinti Paolo Pinton Bilal Piperdi James M Piret Leonidas C Platanias Harald W Platta Edward D Plowey Stefanie Pöggeler Marc Poirot Peter Polčic Angelo Poletti Audrey H Poon Hana Popelka Blagovesta Popova Izabela Poprawa Shibu M Poulose Joanna Poulton Scott K Powers Ted Powers Mercedes Pozuelo-Rubio Krisna Prak Reinhild Prange Mark Prescott Muriel Priault Sharon Prince Richard L Proia Tassula Proikas-Cezanne Holger Prokisch Vasilis J Promponas Karin Przyklenk Rosa Puertollano Subbiah Pugazhenthi Luigi Puglielli Aurora Pujol Julien Puyal Dohun Pyeon Xin Qi Wen-Bin Qian Zheng-Hong Qin Yu Qiu Ziwei Qu Joe Quadrilatero Frederick Quinn Nina Raben Hannah Rabinowich Flavia Radogna Michael J Ragusa Mohamed Rahmani Komal Raina Sasanka Ramanadham Rajagopal Ramesh Abdelhaq Rami Sarron Randall-Demllo Felix Randow Hai Rao V Ashutosh Rao Blake B Rasmussen Tobias M Rasse Edward A Ratovitski Pierre-Emmanuel Rautou Swapan K Ray Babak Razani Bruce H Reed Fulvio Reggiori Markus Rehm Andreas S Reichert Theo Rein David J Reiner Eric Reits Jun Ren Xingcong Ren Maurizio Renna Jane Eb Reusch Jose L Revuelta Leticia Reyes Alireza R Rezaie Robert I Richards Des R Richardson Clémence Richetta Michael A Riehle Bertrand H Rihn Yasuko Rikihisa Brigit E Riley Gerald Rimbach Maria Rita Rippo Konstantinos Ritis Federica Rizzi Elizete Rizzo Peter J Roach Jeffrey Robbins Michel Roberge Gabriela Roca Maria Carmela Roccheri Sonia Rocha Cecilia Mp Rodrigues Clara I Rodríguez Santiago Rodriguez de Cordoba Natalia Rodriguez-Muela Jeroen Roelofs Vladimir V Rogov Troy T Rohn Bärbel Rohrer Davide Romanelli Luigina Romani Patricia Silvia Romano M Isabel G Roncero Jose Luis Rosa Alicia Rosello Kirill V Rosen Philip Rosenstiel Magdalena Rost-Roszkowska Kevin A Roth Gael Roué Mustapha Rouis Kasper M Rouschop Daniel T Ruan Diego Ruano David C Rubinsztein Edmund B Rucker Assaf Rudich Emil Rudolf Ruediger Rudolf Markus A Ruegg Carmen Ruiz-Roldan Avnika Ashok Ruparelia Paola Rusmini David W Russ Gian Luigi Russo Giuseppe Russo Rossella Russo Tor Erik Rusten Victoria Ryabovol Kevin M Ryan Stefan W Ryter David M Sabatini Michael Sacher Carsten Sachse Michael N Sack Junichi Sadoshima Paul Saftig Ronit Sagi-Eisenberg Sumit Sahni Pothana Saikumar Tsunenori Saito Tatsuya Saitoh Koichi Sakakura Machiko Sakoh-Nakatogawa Yasuhito Sakuraba María Salazar-Roa Paolo Salomoni Ashok K Saluja Paul M Salvaterra Rosa Salvioli Afshin Samali Anthony Mj Sanchez José A Sánchez-Alcázar Ricardo Sanchez-Prieto Marco Sandri Miguel A Sanjuan Stefano Santaguida Laura Santambrogio Giorgio Santoni Claudia Nunes Dos Santos Shweta Saran Marco Sardiello Graeme Sargent Pallabi Sarkar Sovan Sarkar Maria Rosa Sarrias Minnie M Sarwal Chihiro Sasakawa Motoko Sasaki Miklos Sass Ken Sato Miyuki Sato Joseph Satriano Niramol Savaraj Svetlana Saveljeva Liliana Schaefer Ulrich E Schaible Michael Scharl Hermann M Schatzl Randy Schekman Wiep Scheper Alfonso Schiavi Hyman M Schipper Hana Schmeisser Jens Schmidt Ingo Schmitz Bianca E Schneider E Marion Schneider Jaime L Schneider Eric A Schon Miriam J Schönenberger Axel H Schönthal Daniel F Schorderet Bernd Schröder Sebastian Schuck Ryan J Schulze Melanie Schwarten Thomas L Schwarz Sebastiano Sciarretta Kathleen Scotto A Ivana Scovassi Robert A Screaton Mark Screen Hugo Seca Simon Sedej Laura Segatori Nava Segev Per O Seglen Jose M Seguí-Simarro Juan Segura-Aguilar Ekihiro Seki Christian Sell Iban Seiliez Clay F Semenkovich Gregg L Semenza Utpal Sen Andreas L Serra Ana Serrano-Puebla Hiromi Sesaki Takao Setoguchi Carmine Settembre John J Shacka Ayesha N Shajahan-Haq Irving M Shapiro Shweta Sharma Hua She C-K James Shen Chiung-Chyi Shen Han-Ming Shen Sanbing Shen Weili Shen Rui Sheng Xianyong Sheng Zu-Hang Sheng Trevor G Shepherd Junyan Shi Qiang Shi Qinghua Shi Yuguang Shi Shusaku Shibutani Kenichi Shibuya Yoshihiro Shidoji Jeng-Jer Shieh Chwen-Ming Shih Yohta Shimada Shigeomi Shimizu Dong Wook Shin Mari L Shinohara Michiko Shintani Takahiro Shintani Tetsuo Shioi Ken Shirabe Ronit Shiri-Sverdlov Orian Shirihai Gordon C Shore Chih-Wen Shu Deepak Shukla Andriy A Sibirny Valentina Sica Christina J Sigurdson Einar M Sigurdsson Puran Singh Sijwali Beata Sikorska Wilian A Silveira Sandrine Silvente-Poirot Gary A Silverman Jan Simak Thomas Simmet Anna Katharina Simon Hans-Uwe Simon Cristiano Simone Matias Simons Anne Simonsen Rajat Singh Shivendra V Singh Shrawan K Singh Debasish Sinha Sangita Sinha Frank A Sinicrope Agnieszka Sirko Kapil Sirohi Balindiwe Jn Sishi Annie Sittler Parco M Siu Efthimios Sivridis Anna Skwarska Ruth Slack Iva Slaninová Nikolai Slavov Soraya S Smaili Keiran Sm Smalley Duncan R Smith Stefaan J Soenen Scott A Soleimanpour Anita Solhaug Kumaravel Somasundaram Jin H Son Avinash Sonawane Chunjuan Song Fuyong Song Hyun Kyu Song Ju-Xian Song Wei Song Kai Y Soo Anil K Sood Tuck Wah Soong Virawudh Soontornniyomkij Maurizio Sorice Federica Sotgia David R Soto-Pantoja Areechun Sotthibundhu Maria João Sousa Herman P Spaink Paul N Span Anne Spang Janet D Sparks Peter G Speck Stephen A Spector Claudia D Spies Wolfdieter Springer Daret St Clair Alessandra Stacchiotti Bart Staels Michael T Stang Daniel T Starczynowski Petro Starokadomskyy Clemens Steegborn John W Steele Leonidas Stefanis Joan Steffan Christine M Stellrecht Harald Stenmark Tomasz M Stepkowski Stęphan T Stern Craig Stevens Brent R Stockwell Veronika Stoka Zuzana Storchova Björn Stork Vassilis Stratoulias Dimitrios J Stravopodis Pavel Strnad Anne Marie Strohecker Anna-Lena Ström Per Stromhaug Jiri Stulik Yu-Xiong Su Zhaoliang Su Carlos S Subauste Srinivasa Subramaniam Carolyn M Sue Sang Won Suh Xinbing Sui Supawadee Sukseree David Sulzer Fang-Lin Sun Jiaren Sun Jun Sun Shi-Yong Sun Yang Sun Yi Sun Yingjie Sun Vinod Sundaramoorthy Joseph Sung Hidekazu Suzuki Kuninori Suzuki Naoki Suzuki Tadashi Suzuki Yuichiro J Suzuki Michele S Swanson Charles Swanton Karl Swärd Ghanshyam Swarup Sean T Sweeney Paul W Sylvester Zsuzsanna Szatmari Eva Szegezdi Peter W Szlosarek Heinrich Taegtmeyer Marco Tafani Emmanuel Taillebourg Stephen Wg Tait Krisztina Takacs-Vellai Yoshinori Takahashi Szabolcs Takáts Genzou Takemura Nagio Takigawa Nicholas J Talbot Elena Tamagno Jerome Tamburini Cai-Ping Tan Lan Tan Mei Lan Tan Ming Tan Yee-Joo Tan Keiji Tanaka Masaki Tanaka Daolin Tang Dingzhong Tang Guomei Tang Isei Tanida Kunikazu Tanji Bakhos A Tannous Jose A Tapia Inmaculada Tasset-Cuevas Marc Tatar Iman Tavassoly Nektarios Tavernarakis Allen Taylor Graham S Taylor Gregory A Taylor J Paul Taylor Mark J Taylor Elena V Tchetina Andrew R Tee Fatima Teixeira-Clerc Sucheta Telang Tewin Tencomnao Ba-Bie Teng Ru-Jeng Teng Faraj Terro Gianluca Tettamanti Arianne L Theiss Anne E Theron Kelly Jean Thomas Marcos P Thomé Paul G Thomes Andrew Thorburn Jeremy Thorner Thomas Thum Michael Thumm Teresa Lm Thurston Ling Tian Andreas Till Jenny Pan-Yun Ting Vladimir I Titorenko Lilach Toker Stefano Toldo Sharon A Tooze Ivan Topisirovic Maria Lyngaas Torgersen Liliana Torosantucci Alicia Torriglia Maria Rosaria Torrisi Cathy Tournier Roberto Towns Vladimir Trajkovic Leonardo H Travassos Gemma Triola Durga Nand Tripathi Daniela Trisciuoglio Rodrigo Troncoso Ioannis P Trougakos Anita C Truttmann Kuen-Jer Tsai Mario P Tschan Yi-Hsin Tseng Takayuki Tsukuba Allan Tsung Andrey S Tsvetkov Shuiping Tu Hsing-Yu Tuan Marco Tucci David A Tumbarello Boris Turk Vito Turk Robin Fb Turner Anders A Tveita Suresh C Tyagi Makoto Ubukata Yasuo Uchiyama Andrej Udelnow Takashi Ueno Midori Umekawa Rika Umemiya-Shirafuji Benjamin R Underwood Christian Ungermann Rodrigo P Ureshino Ryo Ushioda Vladimir N Uversky Néstor L Uzcátegui Thomas Vaccari Maria I Vaccaro Libuše Váchová Helin Vakifahmetoglu-Norberg Rut Valdor Enza Maria Valente Francois Vallette Angela M Valverde Greet Van den Berghe Ludo Van Den Bosch Gijs R van den Brink F Gisou van der Goot Ida J van der Klei Luc Jw van der Laan Wouter G van Doorn Marjolein van Egmond Kenneth L van Golen Luc Van Kaer Menno van Lookeren Campagne Peter Vandenabeele Wim Vandenberghe Ilse Vanhorebeek Isabel Varela-Nieto M Helena Vasconcelos Radovan Vasko Demetrios G Vavvas Ignacio Vega-Naredo Guillermo Velasco Athanassios D Velentzas Panagiotis D Velentzas Tibor Vellai Edo Vellenga Mikkel Holm Vendelbo Kartik Venkatachalam Natascia Ventura Salvador Ventura Patrícia St Veras Mireille Verdier Beata G Vertessy Andrea Viale Michel Vidal Helena L A Vieira Richard D Vierstra Nadarajah Vigneswaran Neeraj Vij Miquel Vila Margarita Villar Victor H Villar Joan Villarroya Cécile Vindis Giampietro Viola Maria Teresa Viscomi Giovanni Vitale Dan T Vogl Olga V Voitsekhovskaja Clarissa von Haefen Karin von Schwarzenberg Daniel E Voth Valérie Vouret-Craviari Kristina Vuori Jatin M Vyas Christian Waeber Cheryl Lyn Walker Mark J Walker Jochen Walter Lei Wan Xiangbo Wan Bo Wang Caihong Wang Chao-Yung Wang Chengshu Wang Chenran Wang Chuangui Wang Dong Wang Fen Wang Fuxin Wang Guanghui Wang Hai-Jie Wang Haichao Wang Hong-Gang Wang Hongmin Wang Horng-Dar Wang Jing Wang Junjun Wang Mei Wang Mei-Qing Wang Pei-Yu Wang Peng Wang Richard C Wang Shuo Wang Ting-Fang Wang Xian Wang Xiao-Jia Wang Xiao-Wei Wang Xin Wang Xuejun Wang Yan Wang Yanming Wang Ying Wang Ying-Jan Wang Yipeng Wang Yu Wang Yu Tian Wang Yuqing Wang Zhi-Nong Wang Pablo Wappner Carl Ward Diane McVey Ward Gary Warnes Hirotaka Watada Yoshihisa Watanabe Kei Watase Timothy E Weaver Colin D Weekes Jiwu Wei Thomas Weide Conrad C Weihl Günther Weindl Simone Nardin Weis Longping Wen Xin Wen Yunfei Wen Benedikt Westermann Cornelia M Weyand Anthony R White Eileen White J Lindsay Whitton Alexander J Whitworth Joëlle Wiels Franziska Wild Manon E Wildenberg Tom Wileman Deepti Srinivas Wilkinson Simon Wilkinson Dieter Willbold Chris Williams Katherine Williams Peter R Williamson Konstanze F Winklhofer Steven S Witkin Stephanie E Wohlgemuth Thomas Wollert Ernst J Wolvetang Esther Wong G William Wong Richard W Wong Vincent Kam Wai Wong Elizabeth A Woodcock Karen L Wright Chunlai Wu Defeng Wu Gen Sheng Wu Jian Wu Junfang Wu Mian Wu Min Wu Shengzhou Wu William Kk Wu Yaohua Wu Zhenlong Wu Cristina Pr Xavier Ramnik J Xavier Gui-Xian Xia Tian Xia Weiliang Xia Yong Xia Hengyi Xiao Jian Xiao Shi Xiao Wuhan Xiao Chuan-Ming Xie Zhiping Xie Zhonglin Xie Maria Xilouri Yuyan Xiong Chuanshan Xu Congfeng Xu Feng Xu Haoxing Xu Hongwei Xu Jian Xu Jianzhen Xu Jinxian Xu Liang Xu Xiaolei Xu Yangqing Xu Ye Xu Zhi-Xiang Xu Ziheng Xu Yu Xue Takahiro Yamada Ai Yamamoto Koji Yamanaka Shunhei Yamashina Shigeko Yamashiro Bing Yan Bo Yan Xianghua Yan Zhen Yan Yasuo Yanagi Dun-Sheng Yang Jin-Ming Yang Liu Yang Minghua Yang Pei-Ming Yang Peixin Yang Qian Yang Wannian Yang Wei Yuan Yang Xuesong Yang Yi Yang Ying Yang Zhifen Yang Zhihong Yang Meng-Chao Yao Pamela J Yao Xiaofeng Yao Zhenyu Yao Zhiyuan Yao Linda S Yasui Mingxiang Ye Barry Yedvobnick Behzad Yeganeh Elizabeth S Yeh Patricia L Yeyati Fan Yi Long Yi Xiao-Ming Yin Calvin K Yip Yeong-Min Yoo Young Hyun Yoo 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Zhuang Elio Ziparo Christos E Zois Teresa Zoladek Wei-Xing Zong Antonio Zorzano Susu M Zughaier

Autophagy 2016 ;12(1):1-222

kb Emory University, School of Medicine , Department of Microbiology and Immunology , Atlanta , GA , USA.

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October 2016

Cellular and molecular modifier pathways in tauopathies: the big picture from screening invertebrate models.

J Neurochem 2016 Apr 11;137(1):12-25. Epub 2016 Feb 11.

Schaller Research Group at the University of Heidelberg and DKFZ, Proteostasis in Neurodegenerative Disease (B180), German Cancer Research Center, Heidelberg, Germany.

Abnormal tau accumulations were observed and documented in post-mortem brains of patients affected by Alzheimer's disease (AD) long before the identification of mutations in the Microtubule-associated protein tau (MAPT) gene, encoding the tau protein, in a different neurodegenerative disease called Frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17). The discovery of mutations in the MAPT gene associated with FTDP-17 highlighted that dysfunctions in tau alone are sufficient to cause neurodegeneration. Invertebrate models have been diligently utilized in investigating tauopathies, contributing to the understanding of cellular and molecular pathways involved in disease etiology. An important discovery came with the demonstration that over-expression of human tau in Drosophila leads to premature mortality and neuronal dysfunction including neurodegeneration, recapitulating some key neuropathological features of the human disease. The simplicity of handling invertebrate models combined with the availability of a diverse range of experimental resources make these models, in particular Drosophila a powerful invertebrate screening tool. Consequently, several large-scale screens have been performed using Drosophila, to identify modifiers of tau toxicity. The screens have revealed not only common cellular and molecular pathways, but in some instances the same modifier has been independently identified in two or more screens suggesting a possible role for these modifiers in regulating tau toxicity. The purpose of this review is to discuss the genetic modifier screens on tauopathies performed in Drosophila and C. elegans models, and to highlight the common cellular and molecular pathways that have emerged from these studies. Here, we summarize results of tau toxicity screens providing mechanistic insights into pathological alterations in tauopathies. Key pathways or modifiers that have been identified are associated with a broad range of processes including, but not limited to, phosphorylation, cytoskeleton organization, axonal transport, regulation of cellular proteostasis, transcription, RNA metabolism, cell cycle regulation, and apoptosis. We discuss the utility and application of invertebrate models in elucidating the cellular and molecular functions of novel and uncharacterized disease modifiers identified in large-scale screens as well as for investigating the function of genes identified as risk factors in genome-wide association studies from human patients in the post-genomic era. In this review, we combined and summarized several large-scale modifier screens performed in invertebrate models to identify modifiers of tau toxicity. A summary of the screens show that diverse cellular processes are implicated in the modification of tau toxicity. Kinases and phosphatases are the most predominant class of modifiers followed by components required for cellular proteostasis and axonal transport and cytoskeleton elements.
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http://dx.doi.org/10.1111/jnc.13532DOI Listing
April 2016

Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila.

Neurobiol Dis 2015 Feb 8;74:76-88. Epub 2014 Nov 8.

German Center for Neurodegenerative Diseases (DZNE) Tübingen, Germany; Hertie Institute for Clinical Brain Research, Laboratory of Functional Neurogenetics, Tübingen, Germany. Electronic address:

Inclusions containing Fused in Sarcoma (FUS) are found in familial and sporadic cases of the incurable progressive motor neuron disease amyotrophic lateral sclerosis and in a common form of dementia, frontotemporal dementia. Most disease-associated mutations are located in the C-terminal proline-tyrosine nuclear localization sequence (PY-NLS) of FUS and impair its nuclear import. It has been shown in cell culture that the nuclear import of FUS is mediated by transportin, which binds the PY-NLS and the last arginine/glycine/glycine-rich (RGG) domain of FUS. Methylation of this last RGG domain by protein arginine methyltransferases (PRMTs) weakens transportin binding and therefore impairs nuclear translocation of FUS. To investigate the requirements for the nuclear import of FUS in an in vivo model, we generated different transgenic Drosophila lines expressing human FUS wild type (hFUS wt) and two disease-related variants P525L and R495X, in which the NLS is mutated or completely absent, respectively. To rule out effects caused by heterologous hFUS expression, we analysed the corresponding variants for the Drosophila FUS orthologue Cabeza (Caz wt, P398L, Q349X). Expression of these variants in eyes and motor neurons confirmed the PY-NLS-dependent nuclear localization of FUS/Caz and caused neurodegenerative effects. Surprisingly, FUS/Caz toxicity was correlated to the degree of its nuclear localization in this overexpression model. High levels of nuclear FUS/Caz became insoluble and reduced the endogenous Caz levels, confirming FUS autoregulation in Drosophila. RNAi-mediated knockdown of the two transportin orthologues interfered with the nuclear import of FUS/Caz and also enhanced the eye phenotype. Finally, we screened the Drosophila PRMT proteins (DART1-9) and found that knockdown of Dart1 led to a reduction in methylation of hFUS P525L and aggravated its phenotype. These findings show that the molecular mechanisms controlling the nuclear import of FUS/Caz and FUS autoregulation are conserved between humans and Drosophila. In addition to the well-known neurodegenerative effects of FUS loss-of function, our data suggest toxic potential of overexpressed FUS in the nucleus and of insoluble FUS.
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http://dx.doi.org/10.1016/j.nbd.2014.11.003DOI Listing
February 2015

UBE2E ubiquitin-conjugating enzymes and ubiquitin isopeptidase Y regulate TDP-43 protein ubiquitination.

J Biol Chem 2014 Jul 13;289(27):19164-79. Epub 2014 May 13.

From the Graduate School of Cellular and Molecular Neuroscience, University of Tübingen, Tübingen 72076, Germany, Laboratory of Functional Neurogenetics, Department of Neurodegeneration, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany, Laboratory of Functional Neurogenetics, Department of Neurodegeneration and

Trans-activation element DNA-binding protein of 43 kDa (TDP-43) characterizes insoluble protein aggregates in distinct subtypes of frontotemporal lobar degeneration and amyotrophic lateral sclerosis. TDP-43 mediates many RNA processing steps within distinct protein complexes. Here we identify novel TDP-43 protein interactors found in a yeast two-hybrid screen using an adult human brain cDNA library. We confirmed the TDP-43 interaction of seven hits by co-immunoprecipitation and assessed their co-localization in HEK293E cells. As pathological TDP-43 is ubiquitinated, we focused on the ubiquitin-conjugating enzyme UBE2E3 and the ubiquitin isopeptidase Y (UBPY). When cells were treated with proteasome inhibitor, ubiquitinated and insoluble TDP-43 species accumulated. All three UBE2E family members could enhance the ubiquitination of TDP-43, whereas catalytically inactive UBE2E3(C145S) was much less efficient. Conversely, silencing of UBE2E3 reduced TDP-43 ubiquitination. We examined 15 of the 48 known disease-associated TDP-43 mutants and found that one was excessively ubiquitinated. This strong TDP-43(K263E) ubiquitination was further enhanced by proteasomal inhibition as well as UBE2E3 expression. Conversely, UBE2E3 silencing and expression of UBPY reduced TDP-43(K263E) ubiquitination. Moreover, wild-type but not active site mutant UBPY reduced ubiquitination of TDP-43 C-terminal fragments and of a nuclear import-impaired mutant. In Drosophila melanogaster, UBPY silencing enhanced neurodegenerative TDP-43 phenotypes and the accumulation of insoluble high molecular weight TDP-43 and ubiquitin species. Thus, UBE2E3 and UBPY participate in the regulation of TDP-43 ubiquitination, solubility, and neurodegeneration.
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http://dx.doi.org/10.1074/jbc.M114.561704DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4081952PMC
July 2014

Knockdown of Hsc70-5/mortalin induces loss of synaptic mitochondria in a Drosophila Parkinson's disease model.

PLoS One 2013 30;8(12):e83714. Epub 2013 Dec 30.

Junior Research Group Synaptic Plasticity, Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen, Germany.

Mortalin is an essential component of the molecular machinery that imports nuclear-encoded proteins into mitochondria, assists in their folding, and protects against damage upon accumulation of dysfunctional, unfolded proteins in aging mitochondria. Mortalin dysfunction associated with Parkinson's disease (PD) increases the vulnerability of cultured cells to proteolytic stress and leads to changes in mitochondrial function and morphology. To date, Drosophila melanogaster has been successfully used to investigate pathogenesis following the loss of several other PD-associated genes. We generated the first loss-of-Hsc70-5/mortalin-function Drosophila model. The reduction of Mortalin expression recapitulates some of the defects observed in the existing Drosophila PD-models, which include reduced ATP levels, abnormal wing posture, shortened life span, and reduced spontaneous locomotor and climbing ability. Dopaminergic neurons seem to be more sensitive to the loss of mortalin than other neuronal sub-types and non-neuronal tissues. The loss of synaptic mitochondria is an early pathological change that might cause later degenerative events. It precedes both behavioral abnormalities and structural changes at the neuromuscular junction (NMJ) of mortalin-knockdown larvae that exhibit increased mitochondrial fragmentation. Autophagy is concomitantly up-regulated, suggesting that mitochondria are degraded via mitophagy. Ex vivo data from human fibroblasts identifies increased mitophagy as an early pathological change that precedes apoptosis. Given the specificity of the observed defects, we are confident that the loss-of-mortalin model presented in this study will be useful for further dissection of the complex network of pathways that underlie the development of mitochondrial parkinsonism.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0083714PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3875477PMC
September 2014

The kinesin-3, unc-104 regulates dendrite morphogenesis and synaptic development in Drosophila.

Genetics 2013 Sep 14;195(1):59-72. Epub 2013 Jun 14.

Junior Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen, 72076 Tübingen, Germany.

Kinesin-based transport is important for synaptogenesis, neuroplasticity, and maintaining synaptic function. In an anatomical screen of neurodevelopmental mutants, we identified the exchange of a conserved residue (R561H) in the forkhead-associated domain of the kinesin-3 family member Unc-104/KIF1A as the genetic cause for defects in synaptic terminal- and dendrite morphogenesis. Previous structure-based analysis suggested that the corresponding residue in KIF1A might be involved in stabilizing the activated state of kinesin-3 dimers. Herein we provide the first in vivo evidence for the functional importance of R561. The R561H allele (unc-104(bris)) is not embryonic lethal, which allowed us to investigate consequences of disturbed Unc-104 function on postembryonic synapse development and larval behavior. We demonstrate that Unc-104 regulates the reliable apposition of active zones and postsynaptic densities, possibly by controlling site-specific delivery of its cargo. Next, we identified a role for Unc-104 in restraining neuromuscular junction growth and coordinating dendrite branch morphogenesis, suggesting that Unc-104 is also involved in dendritic transport. Mutations in KIF1A/unc-104 have been associated with hereditary spastic paraplegia and hereditary sensory and autonomic neuropathy type 2. However, we did not observe synapse retraction or dystonic posterior paralysis. Overall, our study demonstrates the specificity of defects caused by selective impairments of distinct molecular motors and highlights the critical importance of Unc-104 for the maturation of neuronal structures during embryonic development, larval synaptic terminal outgrowth, and dendrite morphogenesis.
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http://dx.doi.org/10.1534/genetics.113.151639DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3761313PMC
September 2013

Spastic paraplegia mutation N256S in the neuronal microtubule motor KIF5A disrupts axonal transport in a Drosophila HSP model.

PLoS Genet 2012 29;8(11):e1003066. Epub 2012 Nov 29.

Junior Research Group Synaptic Plasticity, Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen, Germany.

Hereditary spastic paraplegias (HSPs) comprise a group of genetically heterogeneous neurodegenerative disorders characterized by spastic weakness of the lower extremities. We have generated a Drosophila model for HSP type 10 (SPG10), caused by mutations in KIF5A. KIF5A encodes the heavy chain of kinesin-1, a neuronal microtubule motor. Our results imply that SPG10 is not caused by haploinsufficiency but by the loss of endogenous kinesin-1 function due to a selective dominant-negative action of mutant KIF5A on kinesin-1 complexes. We have not found any evidence for an additional, more generalized toxicity of mutant Kinesin heavy chain (Khc) or the affected kinesin-1 complexes. Ectopic expression of Drosophila Khc carrying a human SPG10-associated mutation (N256S) is sufficient to disturb axonal transport and to induce motoneuron disease in Drosophila. Neurofilaments, which have been recently implicated in SPG10 disease manifestation, are absent in arthropods. Impairments in the transport of kinesin-1 cargos different from neurofilaments are thus sufficient to cause HSP-like pathological changes such as axonal swellings, altered structure and function of synapses, behavioral deficits, and increased mortality.
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http://dx.doi.org/10.1371/journal.pgen.1003066DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3510046PMC
May 2013

Cell wall amidase AmiC1 is required for cellular communication and heterocyst development in the cyanobacterium Anabaena PCC 7120 but not for filament integrity.

J Bacteriol 2012 Oct 20;194(19):5218-27. Epub 2012 Jul 20.

Interfaculty Institute for Microbiology and Infection Medicine, Organismic Interactions, University of Tübingen, Tübingen, Germany.

Filamentous cyanobacteria of the order Nostocales display typical properties of multicellular organisms. In response to nitrogen starvation, some vegetative cells differentiate into heterocysts, where fixation of N(2) takes place. Heterocysts provide a micro-oxic compartment to protect nitrogenase from the oxygen produced by the vegetative cells. Differentiation involves fundamental remodeling of the gram-negative cell wall by deposition of a thick envelope and by formation of a neck-like structure at the contact site to the vegetative cells. Cell wall-hydrolyzing enzymes, like cell wall amidases, are involved in peptidoglycan maturation and turnover in unicellular bacteria. Recently, we showed that mutation of the amidase homologue amiC2 gene in Nostoc punctiforme ATCC 29133 distorts filament morphology and function. Here, we present the functional characterization of two amiC paralogues from Anabaena sp. strain PCC 7120. The amiC1 (alr0092) mutant was not able to differentiate heterocysts or to grow diazotrophically, whereas the amiC2 (alr0093) mutant did not show an altered phenotype under standard growth conditions. In agreement, fluorescence recovery after photobleaching (FRAP) studies showed a lack of cell-cell communication only in the AmiC1 mutant. Green fluorescent protein (GFP)-tagged AmiC1 was able to complement the mutant phenotype to wild-type properties. The protein localized in the septal regions of newly dividing cells and at the neck region of differentiating heterocysts. Upon nitrogen step-down, no mature heterocysts were developed in spite of ongoing heterocyst-specific gene expression. These results show the dependence of heterocyst development on amidase function and highlight a pivotal but so far underestimated cellular process, the remodeling of peptidoglycan, for the biology of filamentous cyanobacteria.
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http://dx.doi.org/10.1128/JB.00912-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3457231PMC
October 2012

The morphogene AmiC2 is pivotal for multicellular development in the cyanobacterium Nostoc punctiforme.

Mol Microbiol 2011 Mar 10;79(6):1655-69. Epub 2011 Feb 10.

Interfaculty Institute for Microbiology and Infection Medicine, Division Organismic Interactions, University of Tübingen, 72076 Tübingen, Germany.

Filamentous cyanobacteria of the order Nostocales are primordial multicellular organisms, a property widely considered unique to eukaryotes. Their filaments are composed of hundreds of mutually dependent vegetative cells and regularly spaced N(2)-fixing heterocysts, exchanging metabolites and signalling molecules. Furthermore, they may differentiate specialized spore-like cells and motile filaments. However, the structural basis for cellular communication within the filament remained elusive. Here we present that mutation of a single gene, encoding cell wall amidase AmiC2, completely changes the morphology and abrogates cell differentiation and intercellular communication. Ultrastructural analysis revealed for the first time a contiguous peptidoglycan sacculus with individual cells connected by a single-layered septal cross-wall. The mutant forms irregular clusters of twisted cells connected by aberrant septa. Rapid intercellular molecule exchange takes place in wild-type filaments, but is completely abolished in the mutant, and this blockage obstructs any cell differentiation, indicating a fundamental importance of intercellular communication for cell differentiation in Nostoc. AmiC2-GFP localizes in the cell wall with a focus in the cross walls of dividing cells, implying that AmiC2 processes the newly synthesized septum into a functional cell-cell communication structure during cell division. AmiC2 thus can be considered as a novel morphogene required for cell-cell communication, cellular development and multicellularity.
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http://dx.doi.org/10.1111/j.1365-2958.2011.07554.xDOI Listing
March 2011

In vivo imaging of intact Drosophila larvae at sub-cellular resolution.

J Vis Exp 2010 Sep 10(43). Epub 2010 Sep 10.

Junior Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen.

Recent improvements in optical imaging, genetically encoded fluorophores and genetic tools allowing efficient establishment of desired transgenic animal lines have enabled biological processes to be studied in the context of a living, and in some instances even behaving, organism. In this protocol we will describe how to anesthetize intact Drosophila larvae, using the volatile anesthetic desflurane, to follow the development and plasticity of synaptic populations at sub-cellular resolution. While other useful methods to anesthetize Drosophila melanogaster larvae have been previously described, the protocol presented herein demonstrates significant improvements due to the following combined key features: (1) A very high degree of anesthetization; even the heart beat is arrested allowing for lateral resolution of up to 150 nm, (2) a high survival rate of >90% per anesthetization cycle, permitting the recording of more than five time-points over a period of hours to days and (3) a high sensitivity enabling us in 2 instances to study the dynamics of proteins expressed at physiological levels. In detail, we were able to visualize the postsynaptic glutamate receptor subunit GluR-IIA expressed via the endogenous promoter in stable transgenic lines and the exon trap line FasII-GFP. (4) In contrast to other methods the larvae can be imaged not only alive, but also intact (i.e. non-dissected) allowing observation to occur over a number of days. The accompanying video details the function of individual parts of the in vivo imaging chamber, the correct mounting of the larvae, the anesthetization procedure, how to re-identify specific positions within a larva and the safe removal of the larvae from the imaging chamber.
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http://dx.doi.org/10.3791/2249DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3157860PMC
September 2010

Knockdown of transactive response DNA-binding protein (TDP-43) downregulates histone deacetylase 6.

EMBO J 2010 Jan 12;29(1):209-21. Epub 2009 Nov 12.

Laboratory of Functional Neurogenetics, Department of Neurodegeneration, Hertie Institute for Clinical Brain Research, Tübingen, Germany.

TDP-43 is an RNA/DNA-binding protein implicated in transcriptional repression and mRNA processing. Inclusions of TDP-43 are hallmarks of frontotemporal dementia and amyotrophic lateral sclerosis. Besides aggregation of TDP-43, loss of nuclear localization is observed in disease. To identify relevant targets of TDP-43, we performed expression profiling. Thereby, histone deacetylase 6 (HDAC6) downregulation was discovered on TDP-43 silencing and confirmed at the mRNA and protein level in human embryonic kidney HEK293E and neuronal SH-SY5Y cells. This was accompanied by accumulation of the major HDAC6 substrate, acetyl-tubulin. HDAC6 levels were restored by re-expression of TDP-43, dependent on RNA binding and the C-terminal protein interaction domains. Moreover, TDP-43 bound specifically to HDAC6 mRNA arguing for a direct functional interaction. Importantly, in vivo validation in TDP-43 knockout Drosophila melanogaster confirmed the specific downregulation of HDAC6. HDAC6 is necessary for protein aggregate formation and degradation. Indeed, HDAC6-dependent reduction of cellular aggregate formation and increased cytotoxicity of polyQ-expanded ataxin-3 were found in TDP-43 silenced cells. In conclusion, loss of functional TDP-43 causes HDAC6 downregulation and might thereby contribute to pathogenesis.
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http://dx.doi.org/10.1038/emboj.2009.324DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2808372PMC
January 2010

PP2A and GSK-3beta act antagonistically to regulate active zone development.

J Neurosci 2009 Sep;29(37):11484-94

Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

The synapse is composed of an active zone apposed to a postsynaptic cluster of neurotransmitter receptors. Each Drosophila neuromuscular junction comprises hundreds of such individual release sites apposed to clusters of glutamate receptors. Here, we show that protein phosphatase 2A (PP2A) is required for the development of structurally normal active zones opposite glutamate receptors. When PP2A is inhibited presynaptically, many glutamate receptor clusters are unapposed to Bruchpilot (Brp), an active zone protein required for normal transmitter release. These unapposed receptors are not due to presynaptic retraction of synaptic boutons, since other presynaptic components are still apposed to the entire postsynaptic specialization. Instead, these data suggest that Brp localization is regulated at the level of individual release sites. Live imaging of glutamate receptors demonstrates that this disruption to active zone development is accompanied by abnormal postsynaptic development, with decreased formation of glutamate receptor clusters. Remarkably, inhibition of the serine-threonine kinase GSK-3beta completely suppresses the active zone defect, as well as other synaptic morphology phenotypes associated with inhibition of PP2A. These data suggest that PP2A and GSK-3beta function antagonistically to control active zone development, providing a potential mechanism for regulating synaptic efficacy at a single release site.
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http://dx.doi.org/10.1523/JNEUROSCI.5584-08.2009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2776049PMC
September 2009

Activity-dependent site-specific changes of glutamate receptor composition in vivo.

Nat Neurosci 2008 Jun 11;11(6):659-66. Epub 2008 May 11.

Institute for Clinical Neurobiology, Medical Faculty, University of Würzburg, Zinklesweg 10, 97080 Würzburg, Germany.

The subunit composition of postsynaptic non-NMDA-type glutamate receptors (GluRs) determines the function and trafficking of the receptor. Changes in GluR composition have been implicated in the homeostasis of neuronal excitability and synaptic plasticity underlying learning. Here, we imaged GluRs in vivo during the formation of new postsynaptic densities (PSDs) at Drosophila neuromuscular junctions coexpressing GluRIIA and GluRIIB subunits. GluR composition was independently regulated at directly neighboring PSDs on a submicron scale. Immature PSDs typically had large amounts of GluRIIA and small amounts of GluRIIB. During subsequent PSD maturation, however, the GluRIIA/GluRIIB composition changed and became more balanced. Reducing presynaptic glutamate release increased GluRIIA, but decreased GluRIIB incorporation. Moreover, the maturation of GluR composition correlated in a site-specific manner with the level of Bruchpilot, an active zone protein that is essential for mature glutamate release. Thus, we show that an activity-dependent, site-specific control of GluR composition can contribute to match pre- and postsynaptic assembly.
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http://dx.doi.org/10.1038/nn.2122DOI Listing
June 2008

Live imaging of synapse development and measuring protein dynamics using two-color fluorescence recovery after photo-bleaching at Drosophila synapses.

Nat Protoc 2007 ;2(12):3285-98

Department of Cellular Neurology, Hertie Institute for Clinical Brain Research, University of Tübingen, Otfried-Müller Strasse 27, Tübingen D-72076, Germany.

Here we describe how to anesthetize and image Drosophila larvae as to follow 'the life history' of identified synapses and synaptic components. This protocol is sensitive, for example, the distribution of glutamate receptors expressed at physiological levels can be monitored. Typically, 2-20 time points can be recorded in the intact organism. Finally, we discuss how to extract the kinetic information on protein dynamics from two-color fluorescence recovery after photo-bleaching (FRAP) measurements and give advice how to keep the in vivo imager's five arch enemies--limited temporal and spatial resolution, injury of the animal, inactivation of proteins and movement artifacts--in check. While we focus on synapses, as model structure, the protocol can easily be adapted to study other developmental processes such as muscle growth, gut development or tracheal branching.
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http://dx.doi.org/10.1038/nprot.2007.472DOI Listing
February 2008

The Ig cell adhesion molecule Basigin controls compartmentalization and vesicle release at Drosophila melanogaster synapses.

J Cell Biol 2007 Jun;177(5):843-55

Developmental Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.

Synapses can undergo rapid changes in size as well as in their vesicle release function during both plasticity processes and development. This fundamental property of neuronal cells requires the coordinated rearrangement of synaptic membranes and their associated cytoskeleton, yet remarkably little is known of how this coupling is achieved. In a GFP exon-trap screen, we identified Drosophila melanogaster Basigin (Bsg) as an immunoglobulin domain-containing transmembrane protein accumulating at periactive zones of neuromuscular junctions. Bsg is required pre- and postsynaptically to restrict synaptic bouton size, its juxtamembrane cytoplasmic residues being important for that function. Bsg controls different aspects of synaptic structure, including distribution of synaptic vesicles and organization of the presynaptic cortical actin cytoskeleton. Strikingly, bsg function is also required specifically within the presynaptic terminal to inhibit nonsynchronized evoked vesicle release. We thus propose that Bsg is part of a transsynaptic complex regulating synaptic compartmentalization and strength, and coordinating plasma membrane and cortical organization.
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http://dx.doi.org/10.1083/jcb.200701111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2064284PMC
June 2007

Bruchpilot promotes active zone assembly, Ca2+ channel clustering, and vesicle release.

Science 2006 May 13;312(5776):1051-4. Epub 2006 Apr 13.

European Neuroscience Institute Göttingen, Grisebachstrasse 5, 37077 Göttingen, Germany.

The molecular organization of presynaptic active zones during calcium influx-triggered neurotransmitter release is the focus of intense investigation. The Drosophila coiled-coil domain protein Bruchpilot (BRP) was observed in donut-shaped structures centered at active zones of neuromuscular synapses by using subdiffraction resolution STED (stimulated emission depletion) fluorescence microscopy. At brp mutant active zones, electron-dense projections (T-bars) were entirely lost, Ca2+ channels were reduced in density, evoked vesicle release was depressed, and short-term plasticity was altered. BRP-like proteins seem to establish proximity between Ca2+ channels and vesicles to allow efficient transmitter release and patterned synaptic plasticity.
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http://dx.doi.org/10.1126/science.1126308DOI Listing
May 2006

Bruchpilot, a protein with homology to ELKS/CAST, is required for structural integrity and function of synaptic active zones in Drosophila.

Neuron 2006 Mar;49(6):833-44

Lehrstuhl für Genetik und Neurobiologie, Theodor-Boveri-Institut für Biowissenschaften, Am Hubland, D-97074 Würzburg, Germany.

Neurotransmitters are released at presynaptic active zones (AZs). In the fly Drosophila, monoclonal antibody (MAB) nc82 specifically labels AZs. We employ nc82 to identify Bruchpilot protein (BRP) as a previously unknown AZ component. BRP shows homology to human AZ protein ELKS/CAST/ERC, which binds RIM1 in a complex with Bassoon and Munc13-1. The C terminus of BRP displays structural similarities to multifunctional cytoskeletal proteins. During development, transcription of the bruchpilot locus (brp) coincides with neuronal differentiation. Panneural reduction of BRP expression by RNAi constructs permits a first functional characterization of this large AZ protein: larvae show reduced evoked but normal spontaneous transmission at neuromuscular junctions. In adults, we observe loss of T bars at active zones, absence of synaptic components in electroretinogram, locomotor inactivity, and unstable flight (hence "bruchpilot"-crash pilot). We propose that BRP is critical for intact AZ structure and normal-evoked neurotransmitter release at chemical synapses of Drosophila.
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http://dx.doi.org/10.1016/j.neuron.2006.02.008DOI Listing
March 2006

Glutamate receptor dynamics organizing synapse formation in vivo.

Nat Neurosci 2005 Jul;8(7):898-905

European Neuroscience Institute Göttingen, Max-Planck-Society, Waldweg 33, D-37073 Göttingen, Germany.

Insight into how glutamatergic synapses form in vivo is important for understanding developmental and experience-triggered changes of excitatory circuits. Here, we imaged postsynaptic densities (PSDs) expressing a functional, GFP-tagged glutamate receptor subunit (GluR-IIA(GFP)) at neuromuscular junctions of Drosophila melanogaster larvae for several days in vivo. New PSDs, associated with functional and structural presynaptic markers, formed independently of existing synapses and grew continuously until reaching a stable size within hours. Both in vivo photoactivation and photobleaching experiments showed that extrasynaptic receptors derived from diffuse, cell-wide pools preferentially entered growing PSDs. After entering PSDs, receptors were largely immobilized. In comparison, other postsynaptic proteins tested (PSD-95, NCAM and PAK homologs) exchanged faster and with no apparent preference for growing synapses. We show here that new glutamatergic synapses form de novo and not by partitioning processes from existing synapses, suggesting that the site-specific entry of particular glutamate receptor complexes directly controls the assembly of individual PSDs.
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http://dx.doi.org/10.1038/nn1484DOI Listing
July 2005

Four different subunits are essential for expressing the synaptic glutamate receptor at neuromuscular junctions of Drosophila.

J Neurosci 2005 Mar;25(12):3209-18

European Neuroscience Institute Göttingen, Max-Planck-Society, D-37073 Göttingen, Germany.

Three ionotropic glutamate receptor subunits, designated GluRIIA, GluRIIB, and GluRIII, have been identified at neuromuscular junctions of Drosophila. Whereas GluRIIA and GluRIIB are redundant for viability, it was shown recently that GluRIII is essential for both the synaptic localization of GluRIIA and GluRIIB and the viability of Drosophila. Here we identify a fourth and a fifth subunit expressed in the neuromuscular system, which we name GluRIID and GluRIIE. Both new subunits we show to be necessary for survival. Moreover, both GluRIID and GluRIIE are required for the synaptic expression of all other glutamate receptor subunits. All five subunits are interdependent for receptor function, synaptic receptor expression, and viability. This indicates that synaptic glutamate receptors incorporate the GluRIII, GluRIID, and GluRIIE subunit together with either GluRIIA or GluRIIB at the Drosophila neuromuscular junction. At this widely used model synapse, the assembly of four different subunits to form an individual glutamate receptor channel may thus be obligatory. This study opens the way for a further characterization of in vivo glutamate receptor assembly and trafficking using the efficient genetics of Drosophila.
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http://dx.doi.org/10.1523/JNEUROSCI.4194-04.2005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6725071PMC
March 2005