Holographic optogenetic stimulation with calcium imaging as an all-optical tool for cardiac electrophysiological studies (Conference Presentation)

2020 ◽  
Author(s):  
Sebastian Junge ◽  
Felix Schmieder ◽  
Philipp Sasse ◽  
Jürgen W. Czarske ◽  
Maria Leilani Torres-Mapa ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Optogenetic Stimulation ◽  
All Optical ◽  
Electrophysiological Studies

scholarly journals Simultaneous voltage and calcium imaging and optogenetic stimulation with high sensitivity and a wide field of view

2019 ◽  
Vol 10 (2) ◽  
pp. 789 ◽  
Author(s):  
Cuong Nguyen ◽  
Hansini Upadhyay ◽  
Michael Murphy ◽  
Gabriel Borja ◽  
Emily J. Rozsahegyi ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
High Sensitivity ◽  
Field Of View ◽  
Wide Field ◽  
Optogenetic Stimulation ◽  
Wide Field Of View

Holographic optogenetic stimulation with calcium imaging to probe and visualize cardiac activity

2021 ◽  
Author(s):  
Sebastian Junge ◽  
Felix Schmieder ◽  
Phillip Sasse ◽  
Jurgen Czarske ◽  
Maria Leilani Torres-Mapa ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Cardiac Activity ◽  
Optogenetic Stimulation

scholarly journals Neuropeptide VF neurons promote sleep via the serotonergic raphe

2019 ◽  
Author(s):  
Daniel A. Lee ◽  
Grigorios Oikonomou ◽  
Tasha Cammidge ◽  
Young Hong ◽  
David A. Prober
Keyword(s):  
Calcium Imaging ◽  
Neural Circuit ◽  
Raphe Nuclei ◽  
Hypothalamic Neurons ◽  
Optogenetic Stimulation ◽  
Genetic Epistasis ◽  
Normal Sleep ◽  
Genetic Labeling ◽  
Raphé Nuclei ◽  
Stimulation Of

ABSTRACTAlthough several sleep-regulating neurons have been identified, little is known about how they interact with each other for sleep/wake control. We previously identified neuropeptide VF (NPVF) and the hypothalamic neurons that produce it as a sleep-promoting system (Lee et al., 2017). Here we use zebrafish to describe a neural circuit in which neuropeptide VF (npvf)-expressing neurons control sleep via the serotonergic raphe nuclei (RN), a hindbrain structure that promotes sleep in both diurnal zebrafish and nocturnal mice. Using genetic labeling and calcium imaging, we show that npvf-expressing neurons innervate and activate serotonergic RN neurons. We additionally demonstrate that optogenetic stimulation of npvf-expressing neurons induces sleep in a manner that requires NPVF and is abolished when the RN are ablated or lack serotonin. Finally, genetic epistasis demonstrates that NPVF acts upstream of serotonin in the RN to maintain normal sleep levels. These findings reveal a novel hypothalamic-hindbrain circuit for sleep/wake control.

scholarly journals All-optical imaging and patterned stimulation with a one-photon endoscope

2021 ◽  
Author(s):  
Jinyong Zhang ◽  
Ryan N Hughes ◽  
Namsoo Kim ◽  
Isabella P Fallon ◽  
Konstantin I bakhurin ◽  
...  
Keyword(s):  
Neural Activity ◽  
Calcium Imaging ◽  
Neural Circuit ◽  
Integrated System ◽  
Striatal Neurons ◽  
Indirect Pathway ◽  
Cellular Resolution ◽  
All Optical ◽  
Freely Moving

While in vivo calcium imaging makes it possible to record activity in defined neuronal populations with cellular resolution, optogenetics allows selective manipulation of neural activity. Recently, these two tools have been combined to stimulate and record neural activity at the same time, but current approaches often rely on two-photon microscopes that are difficult to use in freely moving animals. To address these limitations, we have developed a new integrated system combining a one-photon endoscope and a digital micromirror device for simultaneous calcium imaging and precise optogenetic photo-stimulation with near cellular resolution (Miniscope with All-optical Patterned Stimulation and Imaging, MAPSI). Using this highly portable system in freely moving mice, we were able to image striatal neurons from either the direct pathway or the indirect pathway while simultaneously activating any neuron of choice in the field of view, or to synthesize arbitrary spatiotemporal patterns of photo-stimulation. We could also select neurons based on their relationship with behavior and recreate the behavior by mimicking the natural neural activity with photo-stimulation. MAPSI thus provides a powerful tool for interrogation of neural circuit function in freely moving animals.

scholarly journals Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors

Science ◽  
2018 ◽  
Vol 360 (6396) ◽  
pp. eaat4422 ◽  
Author(s):  
Tommaso Patriarchi ◽  
Jounhong Ryan Cho ◽  
Katharina Merten ◽  
Mark W. Howe ◽  
Aaron Marley ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Brain Function ◽  
Precise Measurement ◽  
Operating Mode ◽  
Optical Recording ◽  
Spatiotemporal Resolution ◽  
Optogenetic Stimulation ◽  
Mouse Cortex ◽  
Neuronal Imaging ◽  
Induced Changes

Neuromodulatory systems exert profound influences on brain function. Understanding how these systems modify the operating mode of target circuits requires spatiotemporally precise measurement of neuromodulator release. We developed dLight1, an intensity-based genetically encoded dopamine indicator, to enable optical recording of dopamine dynamics with high spatiotemporal resolution in behaving mice. We demonstrated the utility of dLight1 by imaging dopamine dynamics simultaneously with pharmacological manipulation, electrophysiological or optogenetic stimulation, and calcium imaging of local neuronal activity. dLight1 enabled chronic tracking of learning-induced changes in millisecond dopamine transients in mouse striatum. Further, we used dLight1 to image spatially distinct, functionally heterogeneous dopamine transients relevant to learning and motor control in mouse cortex. We also validated our sensor design platform for developing norepinephrine, serotonin, melatonin, and opioid neuropeptide indicators.

scholarly journals Neuropeptide VF neurons promote sleep via the serotonergic raphe

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Daniel A Lee ◽  
Grigorios Oikonomou ◽  
Tasha Cammidge ◽  
Andrey Andreev ◽  
Young Hong ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Genetic Evidence ◽  
Neuronal Circuit ◽  
Hypothalamic Neurons ◽  
Neuronal Populations ◽  
Optogenetic Stimulation ◽  
Normal Sleep ◽  
Genetic Labeling ◽  
Raphé Nuclei ◽  
Stimulation Of

Although several sleep-regulating neuronal populations have been identified, little is known about how they interact with each other to control sleep/wake states. We previously identified neuropeptide VF (NPVF) and the hypothalamic neurons that produce it as a sleep-promoting system (Lee et al., 2017). Here we show using zebrafish that npvf-expressing neurons control sleep via the serotonergic raphe nuclei (RN), a hindbrain structure that is critical for sleep in both diurnal zebrafish and nocturnal mice. Using genetic labeling and calcium imaging, we show that npvf-expressing neurons innervate and can activate serotonergic RN neurons. We also demonstrate that chemogenetic or optogenetic stimulation of npvf-expressing neurons induces sleep in a manner that requires NPVF and serotonin in the RN. Finally, we provide genetic evidence that NPVF acts upstream of serotonin in the RN to maintain normal sleep levels. These findings reveal a novel hypothalamic-hindbrain neuronal circuit for sleep/wake control.

scholarly journals Microfluidics-enabled 96-well perfusion system for high-throughput tissue engineering and long-term all-optical electrophysiology

2020 ◽  
Author(s):  
Lai Wei ◽  
Weizhen Li ◽  
Emilia Entcheva ◽  
Zhenyu Li
Keyword(s):  
High Throughput ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Perfusion System ◽  
Excitable Cells ◽  
Hek Cells ◽  
All Optical ◽  
Electrophysiological Studies ◽  
Induced Pluripotent

ABSTRACTThis work demonstrates a novel high-throughput (HT) microfluidics-enabled uninterrupted perfusion system (HT-μUPS) and validates its use with chronic all-optical electrophysiology in human excitable cells. HT-μUPS consists of a soft multichannel microfluidic plate cover which could button on a commercial HT 96-well plate. Herein, we demonstrate the manufacturing process of the system and its usages in acute and chronic all-optical electrophysiological studies of human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CM) and engineered excitable (Spiking HEK) cells. HT-μUPS perfusion maintained functional voltage and calcium responses in iPSC-CM and Spiking HEK cells under spontaneous conditions and under optogenetic pacing. Long-term culture with HT-μUPS improved cell viability and optogenetically-tracked calcium responses in Spiking HEK cells. The scalability and simplicity of this design and its compatibility with HT all-optical electrophysiology can empower cell-based assays for personalized medicine using patient-derived cells.

scholarly journals A pretectal command system controls hunting behaviour

2019 ◽  
Author(s):  
Paride Antinucci ◽  
Mónica Folgueira ◽  
Isaac H. Bianco
Keyword(s):  
Calcium Imaging ◽  
List Type ◽  
Specific Population ◽  
Predatory Behaviour ◽  
Larval Zebrafish ◽  
Optogenetic Stimulation ◽  
Hunting Behaviour ◽  
Action Sequences ◽  
Innate Behaviour ◽  
Stimulation Of

AbstractFor many species, hunting is an innate behaviour that is crucial for survival, yet the circuits that control predatory action sequences are poorly understood. We used larval zebrafish to identify a command system that controls hunting. By combining calcium imaging with a virtual hunting assay, we identified a discrete pretectal region that is selectively active when animals initiate hunting. Targeted genetic labelling allowed us to examine the function and morphology of individual cells and identify two classes of pretectal neuron that project to ipsilateral optic tectum or the contralateral tegmentum. Optogenetic stimulation of single neurons of either class was able to induce sustained hunting sequences, in the absence of prey. Furthermore, laser ablation of these neurons impaired prey-catching and prevented induction of hunting by optogenetic stimulation of the anterior-ventral tectum. In sum, we define a specific population of pretectal neurons that functions as a command system to drive predatory behaviour.Key findingsPretectal neurons are recruited during hunting initiationOptogenetic stimulation of single pretectal neurons can induce predatory behaviourAblation of pretectal neurons impairs huntingPretectal cells comprise a command system controlling hunting behaviour

scholarly journals Large-scale all-optical dissection of motor cortex connectivity reveals a segregated functional organization of mouse forelimb representations

2021 ◽  
Author(s):  
Francesco Resta ◽  
Elena Montagni ◽  
Giuseppe de Vito ◽  
Alessandro Scaglione ◽  
Anna Letizia Allegra Mascaro ◽  
...  
Keyword(s):  
Motor Cortex ◽  
Large Scale ◽  
Functional Organization ◽  
Voluntary Movements ◽  
Motor Representation ◽  
Activation Patterns ◽  
Optogenetic Stimulation ◽  
All Optical ◽  
The Brain ◽  
Stimulation Of

In rodent motor cortex, the rostral forelimb area (RFA) and the caudal forelimb area (CFA) are major actors in orchestrating the control of forelimb complex movements. However, their intrinsic connections and reciprocal functional organization are still unclear, limiting our understanding of how the brain coordinates and executes voluntary movements. Here we causally probed cortical connectivity and activation patterns triggered by transcranial optogenetic stimulation of ethologically relevant complex movements exploiting a novel large-scale all-optical method in awake mice. Results show specific activation features for each movement class, providing evidence for a segregated functional organization of CFA and RFA. Importantly, we identified a second discrete lateral grasping representation area, namely lateral forelimb area (LFA), with unique connectivity and activation patterns. Therefore, we propose the LFA as a distinct motor representation in the forelimb somatotopic motor map.

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