Organ-on-e-chip: Three-dimensional self-rolled biosensor array for electrical interrogations of human electrogenic spheroids

Cell-cell communication plays a pivotal role in coordination and function of biological systems. Three-dimensional (3D) spheroids provide venues to explore cellular communication for tissue development and drug discovery, as their 3D architecture mimics native in vivo microenvironments. Cellular electrophysiology is a prevalent signaling paradigm for studying electroactive cells. Currently, electrophysiological studies do not provide direct, multisite, simultaneous investigation of tissues in 3D. In this study, 3D self-rolled biosensor arrays (3D-SR-BAs) of either active field-effect transistors or passive microelectrodes were implemented to interface human cardiac spheroids in 3D. The arrays provided continuous and stable multiplexed recordings of field potentials with high sensitivity and spatiotemporal resolution, supported with simultaneous calcium imaging. Our approach enables electrophysiological investigation and monitoring of the complex signal transduction in 3D cellular assemblies toward an organ-on-an-electronic-chip (organ-on-e-chip) platform for tissue maturation investigations and development of drugs for disease treatment, such as arrhythmias.

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PARIS, an optogenetic method for functionally mapping gap junctions

10.1101/465781 ◽

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.

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PARIS, an optogenetic method for functionally mapping gap junctions

eLife ◽

10.7554/elife.43366 ◽

2019 ◽

Vol 8 ◽

Cited By ~ 4

Author(s):

Ling Wu ◽

Ao Dong ◽

Liting Dong ◽

Shi-Qiang Wang ◽

Yulong Li

Keyword(s):

Gap Junctions ◽

Cell Communication ◽

Cell Type Specificity ◽

Spatiotemporal Resolution ◽

Wide Range ◽

Junctional Coupling ◽

Chemical Coupling ◽

Drosophila Brain ◽

And Function

Cell-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 s 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.

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High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

Nature Communications ◽

10.1038/s41467-020-19851-1 ◽

2020 ◽

Vol 11 (1) ◽

Cited By ~ 1

Author(s):

Jiang Lan Fan ◽

Jose A. Rivera ◽

Wei Sun ◽

John Peterson ◽

Henry Haeberle ◽

...

Keyword(s):

Blood Flow ◽

High Speed ◽

Laser Scanning ◽

Three Dimensional ◽

Laser Scanning Microscopy ◽

Fluorescence Signal ◽

Imaging Method ◽

Spatiotemporal Resolution ◽

Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

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Mice with cleavage-resistant N-cadherin exhibit synapse anomaly in the hippocampus and outperformance in spatial learning tasks

Molecular Brain ◽

10.1186/s13041-021-00738-1 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

M. Asada-Utsugi ◽

K. Uemura ◽

M. Kubota ◽

Y. Noda ◽

Y. Tashiro ◽

...

Keyword(s):

Memory Function ◽

Three Dimensional ◽

Excitatory Synapses ◽

Missense Mutations ◽

Glutamatergic Synapses ◽

Learning Tasks ◽

Transmission Electron ◽

Nmdar Activation ◽

And Function

AbstractN-cadherin is a homophilic cell adhesion molecule that stabilizes excitatory synapses, by connecting pre- and post-synaptic termini. Upon NMDA receptor (NMDAR) activation by glutamate, membrane-proximal domains of N-cadherin are cleaved serially by a-disintegrin-and-metalloprotease 10 (ADAM10) and then presenilin 1(PS1, catalytic subunit of the γ-secretase complex). To assess the physiological significance of the initial N-cadherin cleavage, we engineer the mouse genome to create a knock-in allele with tandem missense mutations in the mouse N-cadherin/Cadherin-2 gene (Cdh2R714G, I715D, or GD) that confers resistance on proteolysis by ADAM10 (GD mice). GD mice showed a better performance in the radial maze test, with significantly less revisiting errors after intervals of 30 and 300 s than WT, and a tendency for enhanced freezing in fear conditioning. Interestingly, GD mice reveal higher complexity in the tufts of thorny excrescence in the CA3 region of the hippocampus. Fine morphometry with serial section transmission electron microscopy (ssTEM) and three-dimensional (3D) reconstruction reveals significantly higher synaptic density, significantly smaller PSD area, and normal dendritic spine volume in GD mice. This knock-in mouse has provided in vivo evidence that ADAM10-mediated cleavage is a critical step in N-cadherin shedding and degradation and involved in the structure and function of glutamatergic synapses, which affect the memory function.

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Small footprint optoelectrodes using ring resonators for passive light localization

Microsystems & Nanoengineering ◽

10.1038/s41378-021-00263-0 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Vittorino Lanzio ◽

Gregory Telian ◽

Alexander Koshelev ◽

Paolo Micheletti ◽

Gianni Presti ◽

...

Keyword(s):

State Of The Art ◽

Three Dimensional ◽

Network Activity ◽

Ring Resonators ◽

Spatiotemporal Resolution ◽

Optical Ring Resonators ◽

Light Localization ◽

Order Of Magnitude ◽

Small Footprint

AbstractThe combination of electrophysiology and optogenetics enables the exploration of how the brain operates down to a single neuron and its network activity. Neural probes are in vivo invasive devices that integrate sensors and stimulation sites to record and manipulate neuronal activity with high spatiotemporal resolution. State-of-the-art probes are limited by tradeoffs involving their lateral dimension, number of sensors, and ability to access independent stimulation sites. Here, we realize a highly scalable probe that features three-dimensional integration of small-footprint arrays of sensors and nanophotonic circuits to scale the density of sensors per cross-section by one order of magnitude with respect to state-of-the-art devices. For the first time, we overcome the spatial limit of the nanophotonic circuit by coupling only one waveguide to numerous optical ring resonators as passive nanophotonic switches. With this strategy, we achieve accurate on-demand light localization while avoiding spatially demanding bundles of waveguides and demonstrate the feasibility with a proof-of-concept device and its scalability towards high-resolution and low-damage neural optoelectrodes.

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Titanium oral implants: surface characteristics, interface biology and clinical outcome

Journal of The Royal Society Interface ◽

10.1098/rsif.2010.0118.focus ◽

2010 ◽

Vol 7 (suppl_5) ◽

Cited By ~ 128

Author(s):

Anders Palmquist ◽

Omar M. Omar ◽

Marco Esposito ◽

Jukka Lausmaa ◽

Peter Thomsen

Keyword(s):

Surface Properties ◽

Cellular Response ◽

Randomized Clinical Trials ◽

Three Dimensional ◽

Cell Communication ◽

Surface Characteristics ◽

Titanium Implants ◽

Material Surface ◽

Oral Implants

Bone-anchored titanium implants have revolutionized oral healthcare. Surface properties of oral titanium implants play decisive roles for molecular interactions, cellular response and bone regeneration. Nevertheless, the role of specific surface properties, such as chemical and phase composition and nanoscale features, for the biological in vivo performance remains to be established. Partly, this is due to limited transfer of state-of-the-art preparation techniques to complex three-dimensional geometries, analytical tools and access to minute, intact interfacial layers. As judged by the available results of a few randomized clinical trials, there is no evidence that any particular type of oral implant has superior long-term success. Important insights into the recruitment of mesenchymal stem cells, cell–cell communication at the interface and high-resolution imaging of the interface between the surface oxide and the biological host are prerequisites for the understanding of the mechanisms of osseointegration. Strategies for development of the next generation of material surface modifications for compromised tissue are likely to include time and functionally programmed properties, pharmacological modulation and incorporation of cellular components.

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In vivo three-dimensional molecular imaging with Biosensor Imaging of Redundant Deviation in Shifts (BIRDS) at high spatiotemporal resolution

NMR in Biomedicine ◽

10.1002/nbm.2995 ◽

2013 ◽

Vol 26 (11) ◽

pp. 1589-1595 ◽

Cited By ~ 27

Author(s):

Daniel Coman ◽

Robin A. de Graaf ◽

Douglas L. Rothman ◽

Fahmeed Hyder

Keyword(s):

Molecular Imaging ◽

Three Dimensional ◽

Spatiotemporal Resolution ◽

High Spatiotemporal Resolution

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E11/gp38 Selective Expression in Osteocytes: Regulation by Mechanical Strain and Role in Dendrite Elongation

Molecular and Cellular Biology ◽

10.1128/mcb.02120-05 ◽

2006 ◽

Vol 26 (12) ◽

pp. 4539-4552 ◽

Cited By ~ 170

Author(s):

Keqin Zhang ◽

Cielo Barragan-Adjemian ◽

Ling Ye ◽

Shiva Kotha ◽

Mark Dallas ◽

...

Keyword(s):

Shear Stress ◽

Cell Lines ◽

Mechanical Load ◽

Mechanical Strain ◽

Three Dimensional ◽

Cell Communication ◽

Bone Surface ◽

Cellular Processes ◽

Primary Osteoblasts

ABSTRACT Within mineralized bone, osteocytes form dendritic processes that travel through canaliculi to make contact with other osteocytes and cells on the bone surface. This three-dimensional syncytium is thought to be necessary to maintain viability, cell-to-cell communication, and mechanosensation. E11/gp38 is the earliest osteocyte-selective protein to be expressed as the osteoblast differentiates into an osteoid cell or osteocyte, first appearing on the forming dendritic processes of these cells. Bone extracts contain large amounts of E11, but immunostaining only shows its presence in early osteocytes compared to more deeply embedded cells, suggesting epitope masking by mineral. Freshly isolated primary osteoblasts are negative for E11 expression but begin to express this protein in culture, and expression increases with time, suggesting differentiation into the osteocyte phenotype. Osteoblast-like cell lines 2T3 and Oct-1 also show increased expression of E11 with differentiation and mineralization. E11 is highly expressed in MLO-Y4 osteocyte-like cells compared to osteoblast cell lines and primary osteoblasts. Differentiated, mineralized 2T3 cells and MLO-Y4 cells subjected to fluid flow shear stress show an increase in mRNA for E11. MLO-Y4 cells show an increase in dendricity and elongation of dendrites in response to shear stress that is blocked by small interfering RNA specific to E11. In vivo, E11 expression is also increased by a mechanical load, not only in osteocytes near the bone surface but also in osteocytes more deeply embedded in bone. Maximal expression is observed not in regions of maximal strain but in a region of potential bone remodeling, suggesting that dendrite elongation may be occurring during this process. These data suggest that osteocytes may be able to extend their cellular processes after embedment in mineralized matrix and have implications for osteocytic modification of their microenvironment.

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