Validation of Functional Connectivity of Engineered Neuromuscular Junction With Recombinant Monosynaptic Pseudotyped ΔG-Rabies Virus Tracing

Frontiers in Integrative Neuroscience

Rajeevkumar Raveendran Nair¹  Rosanne van de Wijdeven²  Axel Sandvig⁴  Ulrich Stefan Bauer⁵  Ioanna Sandvig⁵  Vegard Fiskum⁵  Clifford Kentros⁶ 

[1] Centre for Neural Computation, Kavli Institute for Systems Neuroscience, Norwegian University of Science and Technology (NTNU), Trondheim, Norway;Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim, Norway;Department of Clinical Neuroscience, Umeå University Hospital, Umeå, Sweden;Department of Community Medicine and Rehabilitation, Umeå University, Umeå, Sweden;Department of Neuromedicine and Movement Science, Norwegian University of Science and Technology (NTNU), Trondheim, Norway;Institute of Neuroscience, University of Oregon, Eugene, OR, United States;

关键词: amyotrophic lateral sclerosis (ALS);    motor neurons (MNs);    pseudo-organoids;    neuroengineering;    in vitro modeling;   

DOI  :  10.3389/fnint.2022.855071

来源: DOAJ

【 摘 要 】

Current preclinical models of neurodegenerative disease, such as amyotrophic lateral sclerosis (ALS), can significantly benefit from in vitro neuroengineering approaches that enable the selective study and manipulation of neurons, networks, and functional units of interest. Custom-designed compartmentalized microfluidic culture systems enable the co-culture of different relevant cell types in interconnected but fluidically isolated microenvironments. Such systems can thus be applied for ALS disease modeling, as they enable the recapitulation and study of neuromuscular junctions (NMJ) through co-culturing of motor neurons and muscle cells in separate, but interconnected compartments. These in vitro systems are particularly relevant for investigations of mechanistic aspects of the ALS pathological cascade in engineered NMJ, as progressive loss of NMJ functionality may constitute one of the hallmarks of disease related pathology at early onset, in line with the dying back hypothesis. In such models, ability to test whether motor neuron degeneration in ALS starts at the nerve terminal or at the NMJ and retrogradely progresses to the motor neuron cell body largely relies on robust methods for verification of engineered NMJ functionality. In this study, we demonstrate the functionality of engineered NMJs within a microfluidic chip with a differentially perturbable microenvironment using a designer pseudotyped ΔG-rabies virus for retrograde monosynaptic tracing.

【 授权许可】

Unknown