scholarly journals Long-term all-optical interrogation of cortical neurons in awake-behaving non-human primates

2018 ◽  
Author(s):  
Niansheng Ju ◽  
Rundong Jiang ◽  
Stephen L. Macknik ◽  
Susana Martinez-Conde ◽  
Shiming Tang
Keyword(s):  
Cortical Neurons ◽  
Single Photon ◽  
Rhesus Macaques ◽  
Single Cells ◽  
Neuronal Populations ◽  
Two Photon ◽  
Calcium Indicators ◽  
All Optical ◽  
High Level

ABSTRACTWhereas optogenetic techniques have proven successful in their ability to manipulate neuronal populations in species ranging from insects to rodents—with high spatial and temporal fidelity—significant obstacles remain in their application to non-human primates (NHPs). Robust optogenetics-activated behavior and long-term noninvasive monitoring of target neurons, have been challenging in NHPs. Here we present a method for all-optical interrogation (AOI), integrating optical stimulation and simultaneous two-photon imaging of neuronal populations in the primary visual cortex (V1) of awake rhesus macaques. A red-shifted channel-rhodopsin transgene (C1V1) and genetically-encoded calcium indicators (GCaMP5 or GCaMP6s) were delivered by AAVs, and subsequently expressed in V1 neuronal populations for months with high stability. We achieved optogenetic stimulation using both single-photon (1P) activation of neuronal populations and two-photon (2P) activation of single-cells, while simultaneously recording 2P calcium imaging in awake monkeys. Optogenetic manipulations of V1 neuronal populations produced reliable artificial visual percepts. Together, our advances show the feasibility of precise and stable all-optical interrogation of cortical neurons in awake NHPs, which may facilitate broad applications in high-level cognition and pre-clinical testing studies.

scholarly journals Quantum Storage of Single-Photon and Two-Photon Fock States with an All-Optical Quantum Memory

2019 ◽  
Vol 122 (21) ◽  
Author(s):  
M. Bouillard ◽  
G. Boucher ◽  
J. Ferrer Ortas ◽  
B. Pointard ◽  
R. Tualle-Brouri
Keyword(s):  
Single Photon ◽  
Quantum Memory ◽  
Fock States ◽  
Two Photon ◽  
All Optical

scholarly journals Global enhancement of cortical excitability following coactivation of large neuronal populations

2020 ◽  
Vol 117 (33) ◽  
pp. 20254-20264 ◽  
Author(s):  
Deng Zhang ◽  
Xingjian Yan ◽  
Liang She ◽  
Yunqing Wen ◽  
Mu-ming Poo
Keyword(s):  
Cortical Neurons ◽  
Neuronal Excitability ◽  
Cortical Excitability ◽  
Large Population ◽  
Neuronal Firing ◽  
Enhancement Effect ◽  
Neuronal Populations ◽  
Mouse Cortex ◽  
The Brain

Correlated activation of cortical neurons often occurs in the brain and repetitive correlated neuronal firing could cause long-term modifications of synaptic efficacy and intrinsic excitability. We found that repetitive optogenetic activation of neuronal populations in the mouse cortex caused enhancement of optogenetically evoked firing of local coactivated neurons as well as distant cortical neurons in both ipsilateral and contralateral hemispheres. This global enhancement of evoked responses required coactivation of a sufficiently large population of neurons either within one cortical area or distributed in several areas. Enhancement of neuronal firing was saturable after repeated episodes of coactivation, diminished by inhibition ofN-methyl-d-aspartic acid receptors, and accompanied by elevated excitatory postsynaptic potentials, all consistent with activity-induced synaptic potentiation. Chemogenetic inhibition of neuronal activity of the thalamus decreased the enhancement effect, suggesting thalamic involvement. Thus, correlated excitation of large neuronal populations leads to global enhancement of neuronal excitability.

Single-dose AAV-based vaccine induces a high level of neutralizing antibodies and provides long-term protection against SARS-CoV-2 in rhesus macaques

2021 ◽  
Author(s):  
Dali Tong ◽  
Mei Zhang ◽  
Yunru Yang ◽  
Han Xia ◽  
Haiyang Tong ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Prevention And Control ◽  
Rhesus Macaques ◽  
Single Shot ◽  
Receptor Binding Domain ◽  
Global Public Health ◽  
Adeno Associated Virus ◽  
High Level ◽  
And Control

Coronavirus disease 2019 (COVID-19), which is triggered by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, continues to threaten global public health. Developing a vaccine that only requires single immunization but provides long-term protection for the prevention and control of COVID-19 is important. Here, we developed an adeno-associated virus (AAV)-based vaccine expressing a stable receptor-binding domain (SRBD) protein. The vaccine requires only single shot, but provides effective neutralizing antibodies (NAbs) over 300 days in rhesus macaques (Macaca mulatta). In addition, the NAbs are at much higher levels than seen in the sera of convalescent patients. It is worth to note that though we detected the pre-existing AAV2/9 NAbs before immunization, the vaccine still induced high and effective NAbs, and did not boost the AAV2/9 NAbs levels in rhesus macaques. Importantly, AAV-SRBD immune sera efficiently neutralized the SARS-CoV-2 P.1/P.2, B.1.1.7, and B.1.351 variants. Together, all the data suggest the vaccine has great potential in preventing the spread of SARS-CoV-2.

scholarly journals Long-Term Two-Photon Calcium Imaging of Neuronal Populations with Subcellular Resolution in Adult Non-human Primates

Cell Reports ◽  
2015 ◽  
Vol 13 (9) ◽  
pp. 1989-1999 ◽  
Author(s):  
Osamu Sadakane ◽  
Yoshito Masamizu ◽  
Akiya Watakabe ◽  
Shin-Ichiro Terada ◽  
Masanari Ohtsuka ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Neuronal Populations ◽  
Two Photon ◽  
Subcellular Resolution

scholarly journals All-optical interrogation of a direction selective retinal circuit by holographic wave front shaping

2019 ◽  
Author(s):  
G.L.B Spampinato ◽  
E. Ronzitti ◽  
V. Zampini ◽  
U. Ferrari ◽  
F. Trapani ◽  
...  
Keyword(s):  
Bipolar Cell ◽  
Ganglion Cell Layer ◽  
Ganglion Cells ◽  
Single Cells ◽  
Direction Selectivity ◽  
Retinal Neurons ◽  
Two Photon ◽  
Light Levels ◽  
All Optical ◽  
Dim Light

AbstractDirection selective (DS) ganglion cells (GC) in the retina maintain their tuning across a broad range of light levels. Yet very different circuits can shape their responses from bright to dim light, and their respective contributions are difficult to tease apart. In particular, the contribution of the rod bipolar cell (RBC) primary pathway, a key player in dim light, is unclear. To understand its contribution to DSGC response, we designed an all-optical approach allowing precise manipulation of single retinal neurons. Our system activates single cells in the bipolar cell (BC) layer by two-photon (2P) temporally focused holographic illumination, while recording the activity in the ganglion cell layer by 2P Ca2 imaging. By doing so, we demonstrate that RBCs provide an asymmetric input to DSGCs, suggesting they contribute to their direction selectivity. Our results suggest that every circuit providing an input to direction selective cells can generate direction selectivity by itself. This hints at a general principle to achieve robust selectivity in sensory areas.

scholarly journals Single cell in vivo brain optogenetic stimulation by two-photon excitation fluorescence transfer

2020 ◽  
Author(s):  
Lei Tong ◽  
Peng Yuan ◽  
Minggang Chen ◽  
Fuyi Chen ◽  
Joerg Bewersdorf ◽  
...  
Keyword(s):  
Single Cell ◽  
Cross Sections ◽  
Single Photon ◽  
Photon Absorption ◽  
Two Photon ◽  
Photon Excitation ◽  
All Optical ◽  
Stimulation Efficiency ◽  
And Control

AbstractOptogenetics at single-cell resolution can be achieved by two-photon stimulation; however, this requires intense or holographic illumination. We markedly improve stimulation efficiency by positioning fluorophores with high two-photon cross-sections adjacent to opsins. The two-photon-excited fluorescence matches the opsin absorbance and can stimulate opsins in a highly localized manner through efficient single-photon absorption. This indirect fluorescence transfer illumination allows experiments difficult to implement in the live brain such as all-optical neural interrogation and control of regional cerebral blood flow.

scholarly journals Long-term all-optical interrogation of cortical neurons in awake-behaving nonhuman primates

PLoS Biology ◽  
2018 ◽  
Vol 16 (8) ◽  
pp. e2005839 ◽  
Author(s):  
Niansheng Ju ◽  
Rundong Jiang ◽  
Stephen L. Macknik ◽  
Susana Martinez-Conde ◽  
Shiming Tang
Keyword(s):  
Cortical Neurons ◽  
Nonhuman Primates ◽  
All Optical

scholarly journals Ontogeny stage-independent and high-level clonal expansion in vitro of mouse hematopoietic stem cells stimulated by an engineered NUP98-HOX fusion transcription factor

Blood ◽  
2011 ◽  
Vol 118 (16) ◽  
pp. 4366-4376 ◽  
Author(s):  
Sanja Sekulovic ◽  
Maura Gasparetto ◽  
Véronique Lecault ◽  
Corinne A. Hoesli ◽  
David G. Kent ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Single Cells ◽  
Hematopoietic Stem ◽  
Image Tracking ◽  
Large Numbers ◽  
High Level

Abstract Achieving high-level expansion of hematopoietic stem cells (HSCs) in vitro will have an important clinical impact in addition to enabling elucidation of their regulation. Here, we couple the ability of engineered NUP98-HOXA10hd expression to stimulate > 1000-fold net expansions of murine HSCs in 10-day cultures initiated with bulk lin−Sca-1+c-kit+ cells, with strategies to purify fetal and adult HSCs and analyze their expansion clonally. We find that NUP98-HOXA10hd stimulates comparable expansions of HSCs from both sources at ∼ 60% to 90% unit efficiency in cultures initiated with single cells. Clonally expanded HSCs consistently show balanced long-term contributions to the lymphoid and myeloid lineages without evidence of leukemogenic activity. Although effects on fetal and adult HSCs were indistinguishable, NUP98-HOXA10hd–transduced adult HSCs did not thereby gain a competitive advantage in vivo over freshly isolated fetal HSCs. Live-cell image tracking of single transduced HSCs cultured in a microfluidic device indicates that NUP98-HOXA10hd does not affect their proliferation kinetics, and flow cytometry confirmed the phenotype of normal proliferating HSCs and allowed reisolation of large numbers of expanded HSCs at a purity of 25%. These findings point to the effects of NUP98-HOXA10hd on HSCs in vitro being mediated by promoting self-renewal and set the stage for further dissection of this process.

scholarly journals Navigating the translational roadblock: Towards highly specific and effective all-optical interrogations of neural circuits

2020 ◽  
Author(s):  
Ting Fu ◽  
Isabelle Arnoux ◽  
Jan Döring ◽  
Hirofumi Watari ◽  
Ignas Stasevicius ◽  
...  
Keyword(s):  
Visual Cortex ◽  
Calcium Imaging ◽  
Cortical Neurons ◽  
Activity Patterns ◽  
Detection Algorithm ◽  
Two Photon ◽  
All Optical ◽  
Laser Sources ◽  
Mouse Visual Cortex

AbstractTwo-photon (2-P) all-optical approaches combine in vivo 2-P calcium imaging and 2-P optogenetic modulations and have the potential to build a framework for network-based therapies, e.g. for rebalancing maladaptive activity patterns in preclinical models of neurological disorders. Here, our goal was to tailor these approaches for this purpose: Firstly, we combined in vivo juxtacellular recordings and GCaMP6f-based 2-P calcium imaging in layer II/III of mouse visual cortex to tune our detection algorithm towards a 100 % specific identification of AP-related calcium transients. False-positive-free detection was achieved at a sensitivity of approximately 73 %. To further increase specificity, secondly, we minimized photostimulation artifacts as a potential source for false-positives by using extended-wavelength-spectrum laser sources for optogenetic stimulation of the excitatory opsin C1V1. We achieved artifact-free all-optical experiments performing photostimulations at 1100 nm or higher and simultaneous calcium imaging at 920 nm in mouse visual cortex in vivo. Thirdly, we determined the spectral range for maximizing efficacy of optogenetic control by performing 2-P photostimulations of individual neurons with wavelengths up to 1300 nm. The rate of evoked transients in GCaMP6f/C1V1-co-expressing cortical neurons peaked already at 1100 nm. By refining spike detection and defining 1100 nm as the optimal wavelength for artifact-free and effective stimulations of C1V1 in GCaMP-based all-optical interrogations, we increased the translational value of these approaches, e.g. for the use in preclinical applications of network-based therapies.One Sentence SummaryWe maximize translational relevance of 2-P all-optical physiology by increasing specificity, minimizing artifacts and optimizing stimulation efficacy.

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