Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip

Sujin Hyung; Seung Ryeol Lee; Yeon Jee Kim; Seokyoung Bang; Dongha Tahk; Jong Chul Park; Jun Kyo Francis Suh; Noo Li Jeon

doi:10.1002/bit.27083

Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip

Sujin Hyung, Seung Ryeol Lee, Yeon Jee Kim, Seokyoung Bang, Dongha Tahk, Jong Chul Park, Jun Kyo Francis Suh, Noo Li Jeon

Department of Medical Engineering

Research output: Contribution to journal › Article › peer-review

15 Citations (Scopus)

Abstract

Axonal regeneration and remyelination of peripheral motor neurons (MNs) are critical for restoring neuromuscular motor function after injury or peripheral neuropathy. We examined whether optogenetically mediated light stimulation (OMLS) could enhance the axon outgrowth and myelination of MNs using three-dimensional motor neuron–Schwann cell (MN–SC) coculture on a microfluidic biochip. The biochip was designed to allow SCs to interact with the axons of MNs, while preventing direct contact between SCs and the cell bodies of MNs. Following coculture with SCs on the microfluidic biochip, MNs were transfected with a light-sensitive channelrhodopsin gene. Transfected MNs subjected to repeated light stimulation (20 Hz, 1 hr) produced significantly longer axons than nontransfected MNs. OMLS of MNs greatly increased the number of myelin basic protein (MBP)-expressing SCs, promoting the initiation of myelination of MNs. Ultrastructurally, OMLS of MNs markedly enhanced the thickness of the compact myelin sheath around the MN axons such that the average thickness was closer to that of the theoretical estimates in vivo. Thus, the MN–SC coculture model on a microfluidic biochip augmented by OMLS of MNs is a feasible platform for studying the relationship of neuronal activity with regrowth and remyelination.

Original language	English
Pages (from-to)	2425-2438
Number of pages	14
Journal	Biotechnology and Bioengineering
Volume	116
Issue number	10
DOIs	https://doi.org/10.1002/bit.27083
Publication status	Published - 2019 Oct 1

Bibliographical note

Funding Information:
The CatCh‐EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP‐09‐04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF‐2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310).

Funding Information:
National Agenda Project of Korea National Research Council of Science & Technology, Grant/Award Numbers: 09‐04, 2N38341; National Research Foundation of Korea, Grant/Award Number: 2018R1A2A1A05019550; Institutional Grant from KIST, Grant/Award Number: 2N38341; Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University, Grant/ Award Number: F14SN02D1310

Funding Information:
The CatCh-EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP-09-04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF-2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310).

Publisher Copyright:
© 2019 Wiley Periodicals, Inc.

All Science Journal Classification (ASJC) codes

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1002/bit.27083

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Hyung, S., Lee, S. R., Kim, Y. J., Bang, S., Tahk, D., Park, J. C., Suh, J. K. F., & Jeon, N. L. (2019). Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip. Biotechnology and Bioengineering, 116(10), 2425-2438. https://doi.org/10.1002/bit.27083

@article{b656649d108f4647974370d665194430,

title = "Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip",

abstract = "Axonal regeneration and remyelination of peripheral motor neurons (MNs) are critical for restoring neuromuscular motor function after injury or peripheral neuropathy. We examined whether optogenetically mediated light stimulation (OMLS) could enhance the axon outgrowth and myelination of MNs using three-dimensional motor neuron–Schwann cell (MN–SC) coculture on a microfluidic biochip. The biochip was designed to allow SCs to interact with the axons of MNs, while preventing direct contact between SCs and the cell bodies of MNs. Following coculture with SCs on the microfluidic biochip, MNs were transfected with a light-sensitive channelrhodopsin gene. Transfected MNs subjected to repeated light stimulation (20 Hz, 1 hr) produced significantly longer axons than nontransfected MNs. OMLS of MNs greatly increased the number of myelin basic protein (MBP)-expressing SCs, promoting the initiation of myelination of MNs. Ultrastructurally, OMLS of MNs markedly enhanced the thickness of the compact myelin sheath around the MN axons such that the average thickness was closer to that of the theoretical estimates in vivo. Thus, the MN–SC coculture model on a microfluidic biochip augmented by OMLS of MNs is a feasible platform for studying the relationship of neuronal activity with regrowth and remyelination.",

author = "Sujin Hyung and Lee, {Seung Ryeol} and Kim, {Yeon Jee} and Seokyoung Bang and Dongha Tahk and Park, {Jong Chul} and Suh, {Jun Kyo Francis} and Jeon, {Noo Li}",

note = "Funding Information: The CatCh‐EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP‐09‐04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF‐2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310). Funding Information: National Agenda Project of Korea National Research Council of Science & Technology, Grant/Award Numbers: 09‐04, 2N38341; National Research Foundation of Korea, Grant/Award Number: 2018R1A2A1A05019550; Institutional Grant from KIST, Grant/Award Number: 2N38341; Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University, Grant/ Award Number: F14SN02D1310 Funding Information: The CatCh-EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP-09-04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF-2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310). Publisher Copyright: {\textcopyright} 2019 Wiley Periodicals, Inc.",

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doi = "10.1002/bit.27083",

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Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip. / Hyung, Sujin; Lee, Seung Ryeol; Kim, Yeon Jee et al.

In: Biotechnology and Bioengineering, Vol. 116, No. 10, 01.10.2019, p. 2425-2438.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

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AU - Lee, Seung Ryeol

AU - Kim, Yeon Jee

AU - Bang, Seokyoung

AU - Tahk, Dongha

AU - Park, Jong Chul

AU - Suh, Jun Kyo Francis

AU - Jeon, Noo Li

N1 - Funding Information: The CatCh‐EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP‐09‐04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF‐2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310). Funding Information: National Agenda Project of Korea National Research Council of Science & Technology, Grant/Award Numbers: 09‐04, 2N38341; National Research Foundation of Korea, Grant/Award Number: 2018R1A2A1A05019550; Institutional Grant from KIST, Grant/Award Number: 2N38341; Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University, Grant/ Award Number: F14SN02D1310 Funding Information: The CatCh-EYFP plasmid was constructed and provided by Dr. Eun Mi Hwang of the Center for Functional Connectomes of the Korea Institute of Science and Technology, Seoul, Korea. Confocal microscopy imaging was carried out on Carl Zeiss microscopes at the Yonsei Advanced Imaging Center, Yonsei University College of Medicine, Seoul, Korea. This study was supported by the National Agenda Project of the Korea National Research Council of Science & Technology (NAP-09-04 to JKFS), an Institutional Grant from KIST (2N38341 to JKFS), the National Research Foundation of Korea (NRF-2018R1A2A1A05019550 to NLJ), and the Brain Korea 21 Plus Project in the Department of Mechanical and Aerospace Engineering, Seoul National University (F14SN02D1310). Publisher Copyright: © 2019 Wiley Periodicals, Inc.

PY - 2019/10/1

Y1 - 2019/10/1

N2 - Axonal regeneration and remyelination of peripheral motor neurons (MNs) are critical for restoring neuromuscular motor function after injury or peripheral neuropathy. We examined whether optogenetically mediated light stimulation (OMLS) could enhance the axon outgrowth and myelination of MNs using three-dimensional motor neuron–Schwann cell (MN–SC) coculture on a microfluidic biochip. The biochip was designed to allow SCs to interact with the axons of MNs, while preventing direct contact between SCs and the cell bodies of MNs. Following coculture with SCs on the microfluidic biochip, MNs were transfected with a light-sensitive channelrhodopsin gene. Transfected MNs subjected to repeated light stimulation (20 Hz, 1 hr) produced significantly longer axons than nontransfected MNs. OMLS of MNs greatly increased the number of myelin basic protein (MBP)-expressing SCs, promoting the initiation of myelination of MNs. Ultrastructurally, OMLS of MNs markedly enhanced the thickness of the compact myelin sheath around the MN axons such that the average thickness was closer to that of the theoretical estimates in vivo. Thus, the MN–SC coculture model on a microfluidic biochip augmented by OMLS of MNs is a feasible platform for studying the relationship of neuronal activity with regrowth and remyelination.

AB - Axonal regeneration and remyelination of peripheral motor neurons (MNs) are critical for restoring neuromuscular motor function after injury or peripheral neuropathy. We examined whether optogenetically mediated light stimulation (OMLS) could enhance the axon outgrowth and myelination of MNs using three-dimensional motor neuron–Schwann cell (MN–SC) coculture on a microfluidic biochip. The biochip was designed to allow SCs to interact with the axons of MNs, while preventing direct contact between SCs and the cell bodies of MNs. Following coculture with SCs on the microfluidic biochip, MNs were transfected with a light-sensitive channelrhodopsin gene. Transfected MNs subjected to repeated light stimulation (20 Hz, 1 hr) produced significantly longer axons than nontransfected MNs. OMLS of MNs greatly increased the number of myelin basic protein (MBP)-expressing SCs, promoting the initiation of myelination of MNs. Ultrastructurally, OMLS of MNs markedly enhanced the thickness of the compact myelin sheath around the MN axons such that the average thickness was closer to that of the theoretical estimates in vivo. Thus, the MN–SC coculture model on a microfluidic biochip augmented by OMLS of MNs is a feasible platform for studying the relationship of neuronal activity with regrowth and remyelination.

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Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three-dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip

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