Structural/Functional Characteristics of Organotypic Spinal Cord Slices under Conditions of Long-Lasting Culturing
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Cytoarchitecture
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The spinal cord contains more than 20 distinct subclasses of neurons that form well-organized neural circuits capable of sensing the environment and generating motor behavior. Although recent studies have described the efficient in vitro generation of spinal motor neurons, the induction of the spinal cord as a whole tissue has not been achieved. In the present study, we demonstrate three-dimensional (3D) induction of dorsal spinal cord-like tissues from human pluripotent stem cells. Our 3D spinal cord induction (3-DiSC) condition recapitulates patterning of the developing dorsal spinal cord and enables the generation of four types of dorsal interneuron marker-positive cell populations. By activating Shh signaling, intermediate and ventral spinal cord-like tissues are successfully induced. After dissociation of these tissues, somatosensory neurons and spinal motor neurons are detected and express neurotransmitters in an in vivo manner. Our approach provides a useful experimental tool for the analysis of human spinal cord development and will contribute to research on the formation and organization of the spinal cord, and its application to regenerative medicine.
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Scyliorhinus canicula
Catecholaminergic cell groups
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Dorsal root ganglion
Anterior Horn Cell
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Dorsal horn neurons in the spinal cord integrate and relay sensory information to higher brain centers. These neurons are organized in specific laminae and different transcription factors are involved in their specification. The murine homeodomain Gbx1 protein is expressed in the mantle zone of the spinal cord at E12.5-13.5, correlating with the appearance of a discernable dorsal horn around E14 and eventually defining a narrow layer in the dorsal horn around perinatal stages. At postnatal stages, Gbx1 identifies a specific subpopulation of GABAergic neurons in the dorsal spinal cord. We have generated a loss of function mutation for Gbx1 and analyzed its consequences during spinal cord development. Gbx1 (-/-) mice are viable and can reproduce as homozygous null mutants. However, the adult mutant mice display an altered gait during forward movement that specifically affects the hindlimbs. This abnormal gait was evaluated by a series of behavioral tests, indicating that locomotion is impaired, but not muscle strength or motor coordination. Molecular analysis showed that the development of the dorsal horn is not profoundly affected in Gbx1 (-/-) mutant mice. However, analysis of terminal neuronal differentiation revealed that the proportion of GABAergic inhibitory interneurons in the superficial dorsal horn is diminished. Our study unveiled a role for Gbx1 in specifying a subset of GABAergic neurons in the dorsal horn of the spinal cord involved in the control of posterior limb movement.
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Interneuron
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[Objective]To study the effect of nerve growth factor(NGF)on differentiation and proliferation of the embryonic rat spinal cord neurons.[Methods]After NGF was added to embryonic spinal cord neurons from the embryonic rat,the number of survival spinal cord neurons was counted and relative protein content in the cells was tested by light microscopy.[Results]The cultured embryonic spinal cord neurons body were aggrandized,the cell protuberance was prolonged,plenty of neuron fiber formed net work,the number of survival spinal cord neurons and the relative protein content increased,which showed the dose-effect relation.[Conclusion]Certain concentration of NGF(20.0~80.0 ng/ml)can promote the differentiation and proliferation of embryonic spinal cord neurons,and enhance the activity of the neurons.
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In order to know the reiationship between Neurotrophin-4 (NT-4)and neurons of spinal cord of adult cat. Usinging specific NT-4 antiserum,We explored the distribution of NT-4 in the neurons of L 6 Spinal cord in adult cat by immunchistochemistry ABC method. Results: The immunoreactive products of NT-4 were distributed the large neurons of ventrolateral and intermedius zone. The immunoreactive product was mainly located in cytoplasm,and many neurites were stained. In addition,many positive neurons in the dorsal horn were oberseved too. The results demonstrated many neurons in gray matter of spinal cord contain NT-4 It suggests that the role of NT-4 involves in the physiological function of neurons in spinal cord of adult cat.
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Subiculum
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Floor plate
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In order to investigate the effect of glial cell line-derived nerurothophic factor (GDNF) on the neurons of spinal cord and dorsal root ganlia (DRG), we observed the localization of GDNFR-alpha on neurons of spinal cord and DRG of rat in vitro. Results: The positive GDNFR-alpha immunoreactivity was present in the neurons of spinal cord and DRG and glial cells. Conclusion: The present results suggested that GDNF might have some important effects on the neurons of spinal cord and DRG and glial cells in vitro.
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Dorsal root ganglion
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