ORIGINAL RESEARCH article
Front. Neurosci.
Sec. Neural Technology
Volume 19 - 2025 | doi: 10.3389/fnins.2025.1619340
This article is part of the Research TopicBridging Computation, Biophysics, Medicine, and Engineering in Neural CircuitsView all 10 articles
Sensory-Spinal Neuron Co-Culture Platform Enables Analysis of Sensory-Driven Spinal Activation
Provisionally accepted
- 1Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan
- 2Department of Precision Engineering, Faculty of Engineering, The University of Tokyo, Bunkyo, Tokyo, Japan
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Background: Pain plays a crucial role in selecting behaviors essential for survival. Nociceptive stimuli are converted into neuronal signals by dorsal root ganglion (DRG) neurons and transmitted via the spinal cord to the brain, where pain is perceived. Chronic pain, characterized by prolonged nociceptive signaling, significantly reduces the quality of life. Specifically, nociplastic pain arises due to heightened spinal neuronal activity. However, the mechanisms underlying this persistent increase remain unclear, impeding the development of effective treatments. Therefore, the present study aimed to develop an experimental platform to investigate how sensory neuron signals increase spinal neuronal activity. Methods: We developed a specialized microstructure enabling a separate culture of DRG and spinal neurons connected functionally by axons extending through microtunnels. Results: Immunofluorescence staining confirmed precise spatial separation and robust neuronal network formation. Microstructures were integrated with high-density microelectrode arrays to facilitate electrophysiological recordings during co-culture. Optogenetic stimulation of DRG neurons significantly activate the spinal neurons, which are not active spontaneously, and increase synchronous activity by 11.8-fold in the spinal neuronal network. Notably, elevated spinal neuron activity persisted for at least 20 min after stimulation ceased, indicating a prolonged neuronal response. Conclusion: This novel co-culture system provides a powerful tool for elucidating the pathogenic mechanisms underlying chronic pain, potentially guiding future therapeutic strategies.
Keywords: co-culture, Dorsal root ganglion neuron, Spinal neuron, electrical recording, Optogenetic stimulation, microfabrication, high-density microelectrode array
Received: 28 Apr 2025; Accepted: 16 Jun 2025.
Copyright: © 2025 Miyahara, Shimba, Kotani and Jimbo. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Yuki Miyahara, Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan
Kenta Shimba, Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan
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