An optimized fluorescent probe for visualizing glutamate neurotransmission
Jonathan S. Marvin,Bart G. Borghuis,Bart G. Borghuis,Lin Tian,Lin Tian,Joseph Cichon,Mark T. Harnett,Jasper Akerboom,Andrew Gordus,Sabine L. Renninger,Tsai Wen Chen,Cornelia I. Bargmann,Michael B. Orger,Eric R. Schreiter,Jonathan B. Demb,Wen-Biao Gan,S. Andrew Hires,Loren L. Looger +17 more
Reads0
Chats0
TLDR
An intensity-based glutamate-sensing fluorescent reporter with signal-to-noise ratio and kinetics appropriate for in vivo imaging is described and its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems is validated.Abstract:
We describe an intensity-based glutamate-sensing fluorescent reporter (iGluSnFR) with signal-to-noise ratio and kinetics appropriate for in vivo imaging. We engineered iGluSnFR in vitro to maximize its fluorescence change, and we validated its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems. In hippocampal culture, iGluSnFR detected single field stimulus-evoked glutamate release events. In pyramidal neurons in acute brain slices, glutamate uncaging at single spines showed that iGluSnFR responds robustly and specifically to glutamate in situ, and responses correlate with voltage changes. In mouse retina, iGluSnFR-expressing neurons showed intact light-evoked excitatory currents, and the sensor revealed tonic glutamate signaling in response to light stimuli. In worms, glutamate signals preceded and predicted postsynaptic calcium transients. In zebrafish, iGluSnFR revealed spatial organization of direction-selective synaptic activity in the optic tectum. Finally, in mouse forelimb motor cortex, iGluSnFR expression in layer V pyramidal neurons revealed task-dependent single-spine activity during running.read more
Citations
More filters
Journal ArticleDOI
Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance.
Linda Madisen,Aleena R. Garner,Daisuke Shimaoka,Amy S. Chuong,Nathan C. Klapoetke,Lu Li,Alexander van der Bourg,Yusuke Niino,Ladan Egolf,Claudio Monetti,Hong Gu,Maya Mills,Adrian Cheng,Bosiljka Tasic,Thuc Nghi Nguyen,Susan M. Sunkin,Andrea Benucci,Andras Nagy,Atsushi Miyawaki,Fritjof Helmchen,Ruth M. Empson,Thomas Knöpfel,Edward S. Boyden,R. Clay Reid,Matteo Carandini,Hongkui Zeng +25 more
TL;DR: These novel transgenic lines greatly expand the ability to monitor and manipulate neuronal activities with increased specificity, and develop driver and double reporter mouse lines and viral vectors using the Cre/Flp and Cre/Dre double recombinase systems.
Journal ArticleDOI
Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors
Tommaso Patriarchi,Jounhong Ryan Cho,Katharina Merten,Mark W. Howe,Aaron Marley,Wei Hong Xiong,Robert W. Folk,Gerard Joey Broussard,Ruqiang Liang,Min Jee Jang,Haining Zhong,Daniel A. Dombeck,Mark von Zastrow,Axel Nimmerjahn,Viviana Gradinaru,John T. Williams,Lin Tian +16 more
TL;DR: The development and validation of dLight1 is reported, a novel suite of intensity-based genetically encoded dopamine indicators that enables ultrafast optical recording of neuronal dopamine dynamics in behaving mice and permits robust detection of physiologically and behaviorally relevant dopamine transients.
Journal ArticleDOI
Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics.
Jasper Akerboom,Nicole Carreras Calderón,Nicole Carreras Calderón,Nicole Carreras Calderón,Lin Tian,Lin Tian,Sebastian Wabnig,Matthias Prigge,Johan Tolö,Andrew Gordus,Michael B. Orger,Michael B. Orger,Kristen E. Severi,John J. Macklin,Ronak Patel,Stefan R. Pulver,Trevor J. Wardill,Trevor J. Wardill,Elisabeth Fischer,Christina Schüler,Tsai Wen Chen,Karen S. Sarkisyan,Jonathan S. Marvin,Cornelia I. Bargmann,Douglas S. Kim,Sebastian Kügler,Leon Lagnado,Peter Hegemann,Alexander Gottschalk,Eric R. Schreiter,Eric R. Schreiter,Loren L. Looger +31 more
TL;DR: Red, single-wavelength GECIs, “RCaMPs,” engineered from circular permutation of the thermostable red fluorescent protein mRuby are described and 2-color calcium imaging is demonstrated both within the same cell (registering mitochondrial and somatic [Ca2+]) and between two populations of cells: neurons and astrocytes.
Journal ArticleDOI
A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific Detection of Dopamine in Flies, Fish, and Mice.
Fangmiao Sun,Jianzhi Zeng,Miao Jing,Miao Jing,Miao Jing,Jingheng Zhou,Jiesi Feng,Jiesi Feng,Jiesi Feng,Scott F. Owen,Yichen Luo,Funing Li,Huan Wang,Huan Wang,Takashi Yamaguchi,Zihao Yong,Yijing Gao,Wanling Peng,Lizhao Wang,Siyu Zhang,Jiulin Du,Dayu Lin,Min Xu,Anatol C. Kreitzer,Guohong Cui,Yulong Li,Yulong Li,Yulong Li +27 more
TL;DR: The development of genetically encoded GPCR-activation-based-DA (GRABDA) sensors enable spatiotemporally precise measurements of DA dynamics in a variety of model organisms while exhibiting complex behaviors.
Journal ArticleDOI
Glutamate triggers long-distance, calcium-based plant defense signaling.
Masatsugu Toyota,Masatsugu Toyota,Masatsugu Toyota,Dirk Spencer,Satoe Sawai-Toyota,Wang Jiaqi,Tong Zhang,Abraham J.K. Koo,Gregg A. Howe,Simon Gilroy +9 more
TL;DR: It is shown that glutamate is a wound signal in plants, which acts as sensors that convert this signal into an increase in intracellular calcium ion concentration that propagates to distant organs, where defense responses are then induced.
References
More filters
Journal ArticleDOI
The structure of the nervous system of the nematode Caenorhabditis elegans
TL;DR: The structure and connectivity of the nervous system of the nematode Caenorhabditis elegans has been deduced from reconstructions of electron micrographs of serial sections as discussed by the authors.
Journal ArticleDOI
Protein production by auto-induction in high-density shaking cultures
TL;DR: Investigation of factors that affect stability, growth, and induction of T7 expression strains in shaking vessels led to the recognition that sporadic, unintended induction of expression in complex media, previously reported by others, is almost certainly caused by small amounts of lactose.
Journal ArticleDOI
Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracerebral microdialysis
TL;DR: It is suggested that the large increase in the content of extracellular glutamate and aspartate in the hippocampus induced by the ischemic period may be one of the causal factors in the damage to certain neurons observed after ischemia.
Journal ArticleDOI
Excitotoxic cell death
TL;DR: On average, NMDA receptors activation may be able to trigger lethal injury more rapidly than AMPA or kainate receptor activation, perhaps reflecting a greater ability to induce calcium influx and subsequent cellular calcium overload.
Journal ArticleDOI
Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators
Lin Tian,S. Andrew Hires,Tianyi Mao,Daniel Huber,M. Eugenia Chiappe,Sreekanth H. Chalasani,Leopoldo Petreanu,Jasper Akerboom,Sean A McKinney,Sean A McKinney,Eric R. Schreiter,Cornelia I. Bargmann,Vivek Jayaraman,Karel Svoboda,Loren L. Looger +14 more
TL;DR: A single-wavelength GCaMP2-based GECI (GCaMP3) is developed, with increased baseline fluorescence, increased dynamic range and higher affinity for calcium, and long-term imaging in the motor cortex of behaving mice revealed large fluorescence changes in imaged neurons over months.