scholarly journals A genetically encoded fluorescent sensor for rapid and specific in vivo detection of norepinephrine

2018 ◽  
Author(s):  
Jiesi Feng ◽  
Changmei Zhang ◽  
Julieta Lischinsky ◽  
Miao Jing ◽  
Jingheng Zhou ◽  
...  
Keyword(s):  
Fluorescent Sensor ◽  
Brain Slices ◽  
High Specificity ◽  
Physiological Processes ◽  
In Vivo Detection ◽  
Wide Range ◽  
Freely Moving ◽  
Rapid Kinetics ◽  
Biogenic Monoamine

AbstractNorepinephrine (NE) and epinephrine (Epi), two key biogenic monoamine neurotransmitters, are involved in a wide range of physiological processes. However, their precise dynamics and regulation remain poorly characterized, in part due to limitations of available techniques for measuring these molecules in vivo. Here, we developed a family of GPCR Activation-Based NE/Epi (GRABNE) sensors with a 230% peak ΔF/F0 response to NE, good photostability, nanomolar-to-micromolar sensitivities, sub-second rapid kinetics, high specificity to NE vs. dopamine. Viral- or transgenic- mediated expression of GRABNE sensors were able to detect electrical-stimulation evoked NE release in the locus coeruleus (LC) of mouse brain slices, looming-evoked NE release in the midbrain of live zebrafish, as well as optogenetically and behaviorally triggered NE release in the LC and hypothalamus of freely moving mice. Thus, GRABNE sensors are a robust tool for rapid and specific monitoring of in vivo NE/Epi transmission in both physiological and pathological processes.

scholarly journals A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo

2020 ◽  
Author(s):  
Ao Dong ◽  
Kaikai He ◽  
Barna Dudok ◽  
Jordan S Farrell ◽  
Wuqiang Guan ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Basolateral Amygdala ◽  
Fluorescent Sensor ◽  
Brain Slices ◽  
High Specificity ◽  
Cultured Neurons ◽  
Spatiotemporal Resolution ◽  
Wide Range

Endocannabinoids (eCBs) are retrograde neuromodulators that play an important role in a wide range of physiological processes; however, the release and in vivo dynamics of eCBs remain largely unknown, due in part to a lack of suitable probes capable of detecting eCBs with sufficient spatiotemporal resolution. Here, we developed a new eCB sensor called GRABeCB2.0. This genetically encoded sensor consists of the human CB1 cannabinoid receptor fused to circular-permutated EGFP, providing cell membrane trafficking, second-resolution kinetics, high specificity for eCBs, and a robust fluorescence response at physiological eCB concentrations. Using the GRABeCB2.0 sensor, we monitored evoked changes in eCB dynamics in both cultured neurons and acute brain slices. Interestingly, in cultured neurons we also observed spontaneous compartmental eCB transients that spanned a distance of approximately 11 μm, suggesting constrained, localized eCB signaling. Moreover, by expressing GRABeCB2.0 in the mouse brain, we readily observed foot shock-elicited and running-triggered eCB transients in the basolateral amygdala and hippocampus, respectively. Lastly, we used GRABeCB2.0 in a mouse seizure model and observed a spreading wave of eCB release that followed a Ca2+ wave through the hippocampus. Thus, GRABeCB2.0 is a robust new probe for measuring the dynamics of eCB release under both physiological and pathological conditions.

scholarly journals Amyloid- and tau-PET imaging in a familial prion kindred

2018 ◽  
Vol 4 (6) ◽  
pp. e290 ◽  
Author(s):  
David T. Jones ◽  
Ryan A. Townley ◽  
Jonathan Graff-Radford ◽  
Hugo Botha ◽  
David S. Knopman ◽  
...  
Keyword(s):  
Case Series ◽  
High Specificity ◽  
Disease Course ◽  
Therapeutic Trials ◽  
In Vivo Detection ◽  
Pet Tracer ◽  
Fourth Decade ◽  
Tau Pet ◽  
Asymptomatic Individuals

ObjectiveTo study the in vivo binding properties of 18F-AV-1451 (tau-PET) and Pittsburgh compound B (PiB-PET) in a unique kindred with a familial prion disorder known to produce amyloid plaques composed of prion protein alongside Alzheimer disease (AD)–like tau tangles.MethodsA case series of 4 symptomatic family members with the 12-octapeptide repeat insertion in the PRNP gene were imaged with 3T MRI, PiB-PET, and tau-PET in their fourth decade of life.ResultsThere was significant neocortical uptake of the tau-PET tracer in all 4 familial prion cases. However, PiB-PET images did not demonstrate abnormally elevated signal in neocortical or cerebellar regions for any of the patients.ConclusionsIn vivo detection of molecular hallmarks of neurodegenerative diseases will be a prerequisite to well-conducted therapeutic trials. Understanding the in vivo behavior of these PET biomarkers in the setting of various neurodegenerative processes is imperative to their proper use in such trials and for research studies focused on the basic neurobiology of neurodegeneration. This study supports the high specificity of neocortical 18F-AV-1451 binding to AD-like tau and the lack of PiB binding to PrP plaques. It is uncertain how early in the disease course tau pathology appears in the brains of individuals who carry this PRNP gene mutation or how it evolves throughout the disease course, but future longitudinal 18F-AV-1451 imaging of symptomatic and asymptomatic individuals in this kindred will help address these uncertainties.

scholarly journals In vivo detection of optically-evoked opioid peptide release

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Ream Al-Hasani ◽  
Jenny-Marie T Wong ◽  
Omar S Mabrouk ◽  
Jordan G McCall ◽  
Gavin P Schmitz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Opioid Peptide ◽  
Neuronal Circuitry ◽  
Endogenous Peptide ◽  
In Vivo Detection ◽  
Highly Sensitive ◽  
Peptide Release ◽  
Freely Moving ◽  
The Brain

Though the last decade has seen accelerated advances in techniques and technologies to perturb neuronal circuitry in the brain, we are still poorly equipped to adequately dissect endogenous peptide release in vivo. To this end we developed a system that combines in vivo optogenetics with microdialysis and a highly sensitive mass spectrometry-based assay to measure opioid peptide release in freely moving rodents.

scholarly journals In vivo brain activity imaging of interactively locomoting mice

2017 ◽  
Author(s):  
Shigenori Inagaki ◽  
Masakazu Agetsuma ◽  
Shinya Ohara ◽  
Toshio Iijima ◽  
Tetsuichi Wazawa ◽  
...  
Keyword(s):  
Visual Cortex ◽  
Primary Visual Cortex ◽  
Brain Activity ◽  
Field Potential ◽  
Brain Functions ◽  
Wide Range ◽  
Novel Method ◽  
Freely Moving ◽  
Voltage Indicator

AbstractElectrophysiological field potential dynamics have been widely used to investigate brain functions and related psychiatric disorders. Conversely, however, various technical limitations of conventional recording methods have limited its applicability to freely moving subjects, especially when they are in a group and socially interacting with each other. Here, we propose a new method to overcome these technical limitations by introducing a bioluminescent voltage indicator called LOTUS-V. Using our simple and fiber-free recording method, named “SNIPA,” we succeeded in capturing brain activity in freely-locomotive mice, without the need for complicated instruments. This novel method further allowed us to simultaneously record from multiple independently-locomotive animals that were interacting with one another. Further, we successfully demonstrated that the primary visual cortex was activated during the interaction. This methodology will further facilitate a wide range of studies in neurobiology and psychiatry.

scholarly journals PARIS, an optogenetic method for functionally mapping gap junctions

2018 ◽  
Author(s):  
Ling Wu ◽  
Ao Dong ◽  
Liting Dong ◽  
Shi-Qiang Wang ◽  
Yulong Li
Keyword(s):  
Gap Junctions ◽  
Cell Communication ◽  
Cell Type Specificity ◽  
Spatiotemporal Resolution ◽  
Physiological Processes ◽  
Wide Range ◽  
Junctional Coupling ◽  
Chemical Coupling ◽  
And Function

ABSTRACTCell-cell communication via gap junctions regulates a wide range of physiological processes by enabling the direct intercellular electrical and chemical coupling. However, the in vivo distribution and function of gap junctions remain poorly understood, partly due to the lack of non-invasive tools with both cell-type specificity and high spatiotemporal resolution. Here we developed PARIS (pairing actuators and receivers to optically isolate gap junctions), a new fully genetically encoded tool for measuring the cell-specific gap junctional coupling (GJC). PARIS successfully enabled monitoring of GJC in several cultured cell lines under physiologically relevant conditions and in distinct genetically defined neurons in Drosophila brain, with ~10-sec temporal resolution and sub-cellular spatial resolution. These results demonstrate that PARIS is a robust, highly sensitive tool for mapping functional gap junctions and study their regulation in both health and disease.

scholarly journals Wireless, battery-free, and fully implantable electrical neurostimulation in freely moving rodents

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Alex Burton ◽  
Sang Min Won ◽  
Arian Kolahi Sohrabi ◽  
Tucker Stuart ◽  
Amir Amirhossein ◽  
...  
Keyword(s):  
Small Animal ◽  
Platinum Electrodes ◽  
Mechanistic Investigation ◽  
Wide Range ◽  
Rapid Dissemination ◽  
Therapeutic Mechanisms ◽  
Freely Moving ◽  
Control Stimulation

AbstractImplantable deep brain stimulation (DBS) systems are utilized for clinical treatment of diseases such as Parkinson’s disease and chronic pain. However, long-term efficacy of DBS is limited, and chronic neuroplastic changes and associated therapeutic mechanisms are not well understood. Fundamental and mechanistic investigation, typically accomplished in small animal models, is difficult because of the need for chronic stimulators that currently require either frequent handling of test subjects to charge battery-powered systems or specialized setups to manage tethers that restrict experimental paradigms and compromise insight. To overcome these challenges, we demonstrate a fully implantable, wireless, battery-free platform that allows for chronic DBS in rodents with the capability to control stimulation parameters digitally in real time. The devices are able to provide stimulation over a wide range of frequencies with biphasic pulses and constant voltage control via low-impedance, surface-engineered platinum electrodes. The devices utilize off-the-shelf components and feature the ability to customize electrodes to enable broad utility and rapid dissemination. Efficacy of the system is demonstrated with a readout of stimulation-evoked neural activity in vivo and chronic stimulation of the medial forebrain bundle in freely moving rats to evoke characteristic head motion for over 36 days.

scholarly journals SNAP-tagged nanobodies enable reversible optical control of a G protein-coupled receptor via a remotely tethered photoswitchable ligand

2018 ◽  
Author(s):  
Helen Farrants ◽  
Amanda Acosta Ruiz ◽  
Vanessa A. Gutzeit ◽  
Dirk Trauner ◽  
Kai Johnsson ◽  
...  
Keyword(s):  
G Protein ◽  
Drug Targets ◽  
Metabotropic Glutamate Receptor ◽  
Receptor Activation ◽  
Metabotropic Glutamate ◽  
Extracellular Signals ◽  
Physiological Processes ◽  
Wide Range ◽  
G Protein Coupled

AbstractG protein-coupled receptors (GPCRs) mediate the transduction of extracellular signals into complex intracellular responses. Despite their ubiquitous roles in physiological processes and as drug targets for a wide range of disorders, the precise mechanisms of GPCR function at the molecular, cellular, and systems levels remain partially understood. In order to dissect the function of individual receptors subtypes with high spatiotemporal precision, various optogenetic and photopharmacological approaches have been reported that use the power of light for receptor activation and deactivation. Here, we introduce a novel and, to date, most remote way of applying photoswitchable orthogonally remotely-tethered ligands (PORTLs) by using a SNAP-tag fused nanobody. Our nanobody-photoswitch conjugates (NPCs) can be used to target a GFP-fused metabotropic glutamate receptor by either gene-free application of purified complexes or co-expression of genetically encoded nanobodies to yield robust, reversible control of agonist binding and subsequent downstream activation. By harboring and combining the selectivity and flexibility of both nanobodies and self-labelling enzymes, we set the stage for targeting endogenous receptors in vivo.

scholarly journals The role of tissue factor in metastasising, neoangiogenesis and hemostasis in cancer

2019 ◽  
Vol 14 (2) ◽  
pp. 70-85
Author(s):  
T. A. Kovalenko ◽  
M. A. Panteleev ◽  
A. N. Sveshnikova
Keyword(s):  
Cell Migration ◽  
Tissue Factor ◽  
Blood Coagulation ◽  
Target Molecule ◽  
Physiological Processes ◽  
Wide Range ◽  
Oncological Diseases

Tissue factor, being the main initiator of the blood coagulation in vivo, is involved in a number of physiological processes, such as angiogenesis or cell migration. These processes are not only significant for normal physiology, but also play a role in the development and progression of oncological diseases. This review presents data on the structure of tissue factor, its expression in normal conditions and in cancer, its role in thrombosis development associated with cancer, in angiogenesis and in metastasis. The involvement of tissue factor in such a wide range of physiological processes important for the progression of cancer makes it an attractive target molecule for therapy.

scholarly journals A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine

Neuron ◽  
2019 ◽  
Vol 102 (4) ◽  
pp. 745-761.e8 ◽  
Author(s):  
Jiesi Feng ◽  
Changmei Zhang ◽  
Julieta E. Lischinsky ◽  
Miao Jing ◽  
Jingheng Zhou ◽  
...  
Keyword(s):  
Fluorescent Sensor ◽  
In Vivo Detection

scholarly journals Quantum dot/antibody conjugates for in vivo cytometric imaging in mice

2015 ◽  
Vol 112 (5) ◽  
pp. 1350-1355 ◽  
Author(s):  
Hee-Sun Han ◽  
Elisabeth Niemeyer ◽  
Yuhui Huang ◽  
Walid S. Kamoun ◽  
John D. Martin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Quantum Dot ◽  
Single Cells ◽  
High Specificity ◽  
Cell Labeling ◽  
Hydrodynamic Diameter ◽  
Calvarial Bone ◽  
Hematopoietic Stem ◽  
Wide Range

Multiplexed, phenotypic, intravital cytometric imaging requires novel fluorophore conjugates that have an appropriate size for long circulation and diffusion and show virtually no nonspecific binding to cells/serum while binding to cells of interest with high specificity. In addition, these conjugates must be stable and maintain a high quantum yield in the in vivo environments. Here, we show that this can be achieved using compact (∼15 nm in hydrodynamic diameter) and biocompatible quantum dot (QD) -Ab conjugates. We developed these conjugates by coupling whole mAbs to QDs coated with norbornene-displaying polyimidazole ligands using tetrazine–norbornene cycloaddition. Our QD immunoconstructs were used for in vivo single-cell labeling in bone marrow. The intravital imaging studies using a chronic calvarial bone window showed that our QD-Ab conjugates diffuse into the entire bone marrow and efficiently label single cells belonging to rare populations of hematopoietic stem and progenitor cells (Sca1+c-Kit+ cells). This in vivo cytometric technique may be useful in a wide range of structural and functional imaging to study the interactions between cells and between a cell and its environment in intact and diseased tissues.

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