Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network
Abstract Neuromuscular diseases usually manifest as abnormalities involving motor neurons, neuromuscular junctions, and skeletal muscle (SkM) in postnatal stage. Present in vitro models of neuromuscular interactions require a long time and lack neuroglia involvement. Our study aimed to construct rod...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-88292-x |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832571774667063296 |
|---|---|
| author | Haiyang Yu Shangbin Yang Yuanfeng Chen Chuangran Wu Jing Xu Yue Yang Rongjie Wu Yinan Guo Zhen Chen Ying Ding Xiang Zeng Ge Li Yuanhuan Ma Qiujian Zheng Yuanshan Zeng Biqin Lai |
| author_facet | Haiyang Yu Shangbin Yang Yuanfeng Chen Chuangran Wu Jing Xu Yue Yang Rongjie Wu Yinan Guo Zhen Chen Ying Ding Xiang Zeng Ge Li Yuanhuan Ma Qiujian Zheng Yuanshan Zeng Biqin Lai |
| author_sort | Haiyang Yu |
| collection | DOAJ |
| description | Abstract Neuromuscular diseases usually manifest as abnormalities involving motor neurons, neuromuscular junctions, and skeletal muscle (SkM) in postnatal stage. Present in vitro models of neuromuscular interactions require a long time and lack neuroglia involvement. Our study aimed to construct rodent bioengineered spinal cord neural network-skeletal muscle (NN-SkM) assembloids to elucidate the interactions between spinal cord neural stem cells (SC-NSCs) and SkM cells and their biological effects on the development and maturation of postnatal spinal cord motor neural circuits. After coculture with SkM cells, SC-NSCs developed into neural networks (NNs) and exhibited a high proportion of glutamatergic and cholinergic neurons, low proportion of neuroglia and gamma-aminobutyric acidergic neurons, and increased expression of synaptic markers. In NN-SkM assembloids, the acetylcholine receptors of SkM cells were upregulated, generating neuromuscular junction-like structures with NNs. The amplitude and frequency of SkM cell contraction in NN-SkM assembloids were increased by optogenetic and glutamate stimulation and blocked by tetrodotoxin and dizocilpine, respectively, confirming the existence of multisynaptic motor NNs. The coculture process involves the secretion of neurotrophin-3 and insulin growth factor-1 by SkM cells, which activate the related ERK-MAPK and PI3K-AKT signaling pathways in NNs. Inhibition of the ERK-MAPK and PI3K-AKT pathways significantly reduces neuronal differentiation and synaptic maturation of neural cells in NN-SkM assembloids, while also decreasing acetylcholine receptor formation on SkM cells. In brief, NN-SkM assembloids simulate the composition of spinal cord motor NNs and respond to motor regulatory signals, providing an in vitro model for studying postnatal development and maturation of spinal cord motor NNs. |
| format | Article |
| id | doaj-art-1e785a4d852b496885750f62d9b0ed7c |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-1e785a4d852b496885750f62d9b0ed7c2025-02-02T12:18:09ZengNature PortfolioScientific Reports2045-23222025-01-0115111610.1038/s41598-025-88292-xConstruction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal networkHaiyang Yu0Shangbin Yang1Yuanfeng Chen2Chuangran Wu3Jing Xu4Yue Yang5Rongjie Wu6Yinan Guo7Zhen Chen8Ying Ding9Xiang Zeng10Ge Li11Yuanhuan Ma12Qiujian Zheng13Yuanshan Zeng14Biqin Lai15Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical SciencesKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityDepartment of Orthopedics, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical UniversityGuangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical SciencesKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityDepartment of Orthopedics, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityGuangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical SciencesKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityGuangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical SciencesKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityKey Laboratory for Stem Cells and Tissue Engineering Ministry of Education, Guangdong Provincial Key Laboratory of Brain Function and Disease, Institute of Spinal Cord Injury, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen UniversityAbstract Neuromuscular diseases usually manifest as abnormalities involving motor neurons, neuromuscular junctions, and skeletal muscle (SkM) in postnatal stage. Present in vitro models of neuromuscular interactions require a long time and lack neuroglia involvement. Our study aimed to construct rodent bioengineered spinal cord neural network-skeletal muscle (NN-SkM) assembloids to elucidate the interactions between spinal cord neural stem cells (SC-NSCs) and SkM cells and their biological effects on the development and maturation of postnatal spinal cord motor neural circuits. After coculture with SkM cells, SC-NSCs developed into neural networks (NNs) and exhibited a high proportion of glutamatergic and cholinergic neurons, low proportion of neuroglia and gamma-aminobutyric acidergic neurons, and increased expression of synaptic markers. In NN-SkM assembloids, the acetylcholine receptors of SkM cells were upregulated, generating neuromuscular junction-like structures with NNs. The amplitude and frequency of SkM cell contraction in NN-SkM assembloids were increased by optogenetic and glutamate stimulation and blocked by tetrodotoxin and dizocilpine, respectively, confirming the existence of multisynaptic motor NNs. The coculture process involves the secretion of neurotrophin-3 and insulin growth factor-1 by SkM cells, which activate the related ERK-MAPK and PI3K-AKT signaling pathways in NNs. Inhibition of the ERK-MAPK and PI3K-AKT pathways significantly reduces neuronal differentiation and synaptic maturation of neural cells in NN-SkM assembloids, while also decreasing acetylcholine receptor formation on SkM cells. In brief, NN-SkM assembloids simulate the composition of spinal cord motor NNs and respond to motor regulatory signals, providing an in vitro model for studying postnatal development and maturation of spinal cord motor NNs.https://doi.org/10.1038/s41598-025-88292-xNeuromuscular interactionTissue engineeringMotor neural networkSkeletal muscleSpinal cord neural stem cells |
| spellingShingle | Haiyang Yu Shangbin Yang Yuanfeng Chen Chuangran Wu Jing Xu Yue Yang Rongjie Wu Yinan Guo Zhen Chen Ying Ding Xiang Zeng Ge Li Yuanhuan Ma Qiujian Zheng Yuanshan Zeng Biqin Lai Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network Scientific Reports Neuromuscular interaction Tissue engineering Motor neural network Skeletal muscle Spinal cord neural stem cells |
| title | Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| title_full | Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| title_fullStr | Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| title_full_unstemmed | Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| title_short | Construction of a rodent neural network-skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| title_sort | construction of a rodent neural network skeletal muscle assembloid that simulate the postnatal development of spinal cord motor neuronal network |
| topic | Neuromuscular interaction Tissue engineering Motor neural network Skeletal muscle Spinal cord neural stem cells |
| url | https://doi.org/10.1038/s41598-025-88292-x |
| work_keys_str_mv | AT haiyangyu constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT shangbinyang constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yuanfengchen constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT chuangranwu constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT jingxu constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yueyang constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT rongjiewu constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yinanguo constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT zhenchen constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yingding constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT xiangzeng constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT geli constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yuanhuanma constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT qiujianzheng constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT yuanshanzeng constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork AT biqinlai constructionofarodentneuralnetworkskeletalmuscleassembloidthatsimulatethepostnataldevelopmentofspinalcordmotorneuronalnetwork |