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Device Technologies and Biomedical Robotics

Prosthetics, Biomechanics, and Assistive Technology

Neuromuscular junction with a mechanical output

Friday, October 25, 2024

9:30 AM – 9:45 AM EST

Location: Room 327

Presenting Author(s)

Hyegi Min (he/him/his)

Postdoctoral Researcher
University of Illinois at Urbana-Champaign
Savoy, Illinois, United States

Co-Author(s)

JW

Jiaojiao Wang

University of Illinois at Urbana-Champaign, United States
JY

Jaesung You

University of Illinois at Urbana-Champaign, United States
Woong Kim

University of Illinois at Urbana-Champaign, United States
YW

Yue Wang (he/him/his)

PhD student
Northwestern University
Evanston, Illinois, United States
Yu Ding, PhD Candidate

Research Assistant
University of Illinois at Urbana-Champaign
Champaign, Illinois, United States
Jongwon Lim

PhD student
University of Illinois at Urbana-Champaign
Champaign, Illinois, United States
YV

Yurii Vlasov

University of Illinois at Urbana-Champaign, United States
JR

John A. Rogers

Northwestern University, United States

Primary Investigator(s)

Rashid Bashir, PhD

Dean
University of Illinois at Urbana-Champaign
Champaign, Illinois, United States

Co-Author(s)

OA

Onur Aydin

Postdoctoral Fellow
University of Illinois at Urbana-Champaign
Urbana, Illinois, United States
Sehong Kang, MS

Graduate student
University of Illinois Urbana Champaign
Urbana, Illinois, United States
MS

M. Taher A. Saif

Professor
University of Illinois at Urbana-Champaign
Urbana, Illinois, United States
Hk

Hyunjoon kong

Associate Professor
University of Illinois Urbana Champaign, United States
XL

Xiuyuan Li
YH

Yonggang Huang

Introduction: Skeletal muscle is one of the promising power sources to generate mechanical forces from biological metabolism. Electric fields or optics have been employed to activate voltage-gated or light-gated ion channel, evoking contraction mechanism, but their functionalities are limited without an external stimulus. To meet this necessity, there is growing interest in developing neuromuscular junction (NMJ) to provide stimulus unit through synaptic signaling. In this study, we report on development of robust NMJ models by integrating motor neurons and skeletal muscles to demonstrate spontaneous and stimulated modulation of muscle contraction modes.

Materials and

Methods: To prepare a mold for culturing multicellular system, polyethylene glycol diacrylate (PEGDA) was 3D printed using stereolithography with the aid of a photo-initiator. As sources of neuromuscular tissues, C2C12 myoblasts and mouse embryonic stem cells (mESCs) were used to form skeletal muscle and neural organoids, respectively. Grown C2C12 cells were seeded into the mold and differentiated into myotubes. The mESCs were cultured and differentiated into neural organoids, following being seeded next to the muscle tissue in the same mold. Then, synaptic connections formed between neurons and myotubes after co-culturing in supplemented media.

Results, Conclusions, and Discussions: Prepared neuromuscular junctions were characterized by immunostaining, revealing the presence of differentiated skeletal muscles and neural organoids expressed by the myosin heavy chain (MF-20) and beta-III-tubulin (tuj1), respectively. Acetylcholine receptors (AChRs) are imaged by the alpha-bungarotoxin (a-BTX) expression. Neural organoids were observed adhering to myotubes and extending neurites, forming junction interfaces. Genetic analysis via qPCR and mRNA sequencing confirmed upregulation of genes related to acetylcholine receptor clustering and stabilization, muscle development, neurodevelopment, and synaptic function. The formation of junctions was also confirmed by the spontaneous twitching of muscles. This highlighted the coupling of neuronal firing and muscle contraction through NMJ-mediated biological signaling. We will also present detailed characterization of the NMJ including results on LED wireless radio frequency controlled uLED based stimulation of the optogenetically transfected neurons in the NMJ. This study provides insightful results on understanding biological signal transduction between muscles and neurons.

Acknowledgements (Optional):