Compartmental microfluidic system for studying muscle–neuron communication and neuromuscular junction maintenance

Ionescu, Ariel; Zahavi, Eitan Erez; Gradus, Tal; Ben-Yaakov, Keren; Perlson, Eran

doi:10.1016/j.ejcb.2015.11.004

Cited by 93 publications

(89 citation statements)

References 31 publications

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“…Microfluidic chambers were prepared as described before (Ionescu et al, 2016, Maimon et al, 2018. E11.5 -E12.5 spinal cord explants were dissected in cold HBSS and placed into Laminin (3 μg/ml) and…”

Section: Microfluidic Chambersmentioning

confidence: 99%

CRMP2 mediates Sema3F-dependent axon pruning and dendritic spine remodeling

Žiak

Weissova

Jeřábková

et al. 2019

Preprint

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Regulation of axon guidance and pruning of inappropriate synapses by class 3 semaphorins is key to development of neural circuits. Collapsin response mediator protein 2 (CRMP2) has been shown to regulate axon guidance by mediating Semaphorin 3A (Sema3A) signaling and its dysfunction has been linked to schizophrenia, however, nothing is known about its role in the synapse pruning. Here, using newly generated crmp2 -/mice we demonstrate that while CRMP2 has only a moderate effect on Sema3A-dependent axon guidance in vivo, it is essential for Sema3F-dependent axon pruning and dendritic spine remodeling. We first demonstrate that CRMP2 deficiency interferes with Sema3A signaling in compartmentalized neuron cultures and leads to a mild defect in axon guidance in peripheral nerves and corpus callosum. Strikingly, we show that crmp2 -/mice display more prominent defects in dendritic spine pruning and stereotyped axon pruning in hippocampus and visual cortex consistent with impaired Sema3F signaling and with autism spectrum disorder (ASD)rather than schizophrenia-like phenotype. Indeed, we demonstrate that CRMP2 mediates Sema3Finduced axon retraction in primary neurons and that crmp2 -/mice display early postnatal behavioral changes linked to ASD. In conclusion, we demonstrate that CRMP2 is an essential mediator of Sema3F-dependent synapse pruning and its dysfunction in early postnatal stages shares histological and behavioral features of ASD.

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Section: Microfluidic Chambersmentioning

confidence: 99%

CRMP2 mediates Sema3F-dependent axon pruning and dendritic spine remodeling

Žiak

Weissova

Jeřábková

et al. 2019

Preprint

Self Cite

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“…For example, as shown in Figure 6.B, Ionescu et al developed a compartmentalized NSC for study of the neuromuscular junction, which enabled the monitoring of muscle-neuron communication. 71 Berdichevsky et al used a compartmentalized NSC to co-culture cortical and hippocampal neurons separated by microgrooves to monitor the development of neurite pathways. 72 A compartmentalized NSC containing a cell-laden hydrogel component was used by Shin et al to co-culture endothelial and neural progenitor cells in a 3D environment allowing them to study the effect of vasculature on neural progenitor cell differentiation.…”

Section: Classes Of Neural Systems On a Chipmentioning

confidence: 99%

Microphysiological Human Brain and Neural Systems-on-a-Chip: Potential Alternatives to Small Animal Models and Emerging Platforms for Drug Discovery and Personalized Medicine

Haring

Sontheimer

Johnson

2017

Stem Cell Rev and Rep

106

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Translational challenges associated with reductionist modeling approaches, as well as ethical concerns and economic implications associated with small animal testing, drive the need for developing microphysiological neural systems for modeling human neurological diseases, disorders and injuries. Here, we provide a comprehensive review of microphysiological neural systems on a chip (NSCs) for modeling higher order trajectories in the human nervous system. Societal, economic, and national security impacts of neurological diseases, disorders and injuries are highlighted to identify critical NSC application spaces. Hierarchical design and manufacturing of NSCs are discussed with distinction of surface- and bulk-based systems. Three broad NSC classes are identified and reviewed: microfluidic NSCs, compartmentalized NSCs, and hydrogel NSCs. Emerging areas and future directions are highlighted, including the application of 3D printing to design and manufacturing of next-generation NSCs, the use of stem cells for constructing patient-specific NSCs, and the application of human NSCs to ‘personalized neurology’. Technical hurdles and remaining challenges are discussed. This review identifies the state-of-the-art design methodologies, manufacturing approaches, and performance capabilities of NSCs. This work suggests NSCs appear poised to revolutionize the modeling of human neurological diseases, disorders and injuries.

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“…Indeed, a number of studies have demonstrated the applicability of various bioengineering methods for creating muscle-neuron co-cultures, including functional NMJs in vitro. [15][16][17][18][19][20][21] In the present study, we model functional human NMJs by compartmentalized culture of iPS cell-derived MN pseudo-organoids and myotubes in a differentially perturbable multi-nodal microfluidic chip and demonstrate that NMJ function can be validated through monosynaptic retrograde tracing using a designer pseudotyped ΔG-rabies virus (RV).…”

Section: Introductionmentioning

confidence: 99%

Modelling functional human neuromuscular junctions in a differentially-perturbable microfluidic environment, validated through recombinant monosynaptic pseudotyped ΔG-rabies virus tracing

Bauer

Wijdeven

Raveendran

et al. 2019

Preprint

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Sweden Graphical abstractFunctional neuromuscular junction in a microfluidic chip (a) Overview of microfluidic chip. Human iPS cell-derived motor neuron aggregates (spheroids indicated by black arrows) are seeded in the three lateral compartments of the chip, while human myotubes (white arrows) are seeded in the middle compartment. (b) Directed connectivity and retrograde virus tracing.Outgrowing axons (yellow arrow) from the motor neuron aggregate enter the directional axon tunnels (grey rectangles) and form connections with the myotubes (white arrow) within the opposite compartment. Addition of a designer monosynaptic pseudotyped ΔG-rabies virus to the myotube compartment, infects the myotubes (green) expressing an exogenous receptor (TVA) and rabies glycoprotein (G), subsequently making infectious viruses that are retrogradely transported through the motor neuron axons (green arrow) back to the neuronal cell bodies within the aggregate, validating neuromuscular junction functionality. AbstractCompartmentalized microfluidic culture systems provide new perspectives in in vitro disease modelling as they enable co-culture of different relevant cell types in interconnected but fluidically isolated microenvironments. Such systems are thus particularly interesting in the context of in vitro modelling of mechanistic aspects of neurodegenerative diseases such as amyotrophic lateral sclerosis, which progressively affect the function of neuromuscular junctions, as they enable the co-culture of motor neurons and muscle cells in separate, but interconnected compartments. In combination with cell reprogramming technologies for the generation of human (including patient-specific) motor neurons, microfluidic platforms can thus become important research tools in preclinical studies. In this study, we present the application of a microfluidic chip with a differentially-perturbable microenvironment as a platform for establishing functional neuromuscular junctions using human induced pluripotent stem cell derived motor neurons and human myotubes. As a novel approach, we demonstrate the functionality of the platform using a designer pseudotyped ΔG-rabies virus for retrograde monosynaptic tracing.

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Compartmental microfluidic system for studying muscle–neuron communication and neuromuscular junction maintenance

Cited by 93 publications

References 31 publications

CRMP2 mediates Sema3F-dependent axon pruning and dendritic spine remodeling

CRMP2 mediates Sema3F-dependent axon pruning and dendritic spine remodeling

Microphysiological Human Brain and Neural Systems-on-a-Chip: Potential Alternatives to Small Animal Models and Emerging Platforms for Drug Discovery and Personalized Medicine

Modelling functional human neuromuscular junctions in a differentially-perturbable microfluidic environment, validated through recombinant monosynaptic pseudotyped ΔG-rabies virus tracing

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