scholarly journals A Role for the Cannabinoid 1 Receptor in Neuronal Differentiation of Adult Spinal Cord Progenitors in vitro is Revealed through Pharmacological Inhibition and Genetic Deletion

2012 ◽  
Vol 6 ◽  
Author(s):  
Alexandra Sideris ◽  
Tatiana Bekker ◽  
Wai Si Chan ◽  
Jose V. Montoya-Gacharna ◽  
Thomas J. J. Blanck ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Neuronal Differentiation ◽  
Pharmacological Inhibition ◽  
Cannabinoid 1 Receptor ◽  
Genetic Deletion ◽  
Adult Spinal Cord

An in vitro spinal cord slice preparation for recording from lumbar motoneurons of the adult mouse

2012 ◽  
Vol 107 (2) ◽  
pp. 728-741 ◽  
Author(s):  
Pratip Mitra ◽  
Robert M. Brownstone
Keyword(s):  
Spinal Cord ◽  
Fluorescent Protein ◽  
Ethyl Pyruvate ◽  
Postnatal Period ◽  
Slice Preparation ◽  
Spinal Cord Slice ◽  
Whole Cell Patch Clamp ◽  
Firing Properties ◽  
Adult Spinal Cord

The development of central nervous system slice preparations for electrophysiological studies has led to an explosion of knowledge of neuronal properties in health and disease. Studies of spinal motoneurons in these preparations, however, have been largely limited to the early postnatal period, as adult motoneurons are vulnerable to the insults sustained by the preparation. We therefore sought to develop an adult spinal cord slice preparation that permits recording from lumbar motoneurons. To accomplish this, we empirically optimized the composition of solutions used during preparation in order to limit energy failure, reduce harmful ionic fluxes, mitigate oxidative stress, and prevent excitotoxic cell death. In addition to other additives, this involved the use of ethyl pyruvate, which serves as an effective nutrient and antioxidant. We also optimized and incorporated a host of previously published modifications used for other in vitro preparations, such as the use of polyethylene glycol. We provide an in-depth description of the preparation protocol and discuss the rationale underlying each modification. By using this protocol, we obtained stable whole cell patch-clamp recordings from identified fluorescent protein-labeled motoneurons in adult slices; here, we describe the firing properties of these adult motoneurons. We propose that this preparation will allow further studies of how motoneurons integrate activity to produce adult motor behaviors and how pathological processes such as amyotrophic lateral sclerosis affect these neurons.

scholarly journals Ecto-domain phosphorylation promotes functional recovery from spinal cord injury

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Kenji Suehiro ◽  
Yuka Nakamura ◽  
Shuai Xu ◽  
Youichi Uda ◽  
Takafumi Matsumura ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Enhanced Recovery ◽  
Neuronal Cell ◽  
Cell Production ◽  
Locomotor Function ◽  
Neuronal Precursor Cells ◽  
Cord Injury ◽  
Adult Spinal Cord

Abstract Inhibition of Nogo-66 receptor (NgR) can promote recovery following spinal cord injury. The ecto-domain of NgR can be phosphorylated by protein kinase A (PKA), which blocks activation of the receptor. Here, we found that infusion of PKA plus ATP into the damaged spinal cord can promote recovery of locomotor function. While significant elongation of cortical-spinal axons was not detectable even in the rats showing enhanced recovery, neuronal precursor cells were observed in the region where PKA plus ATP were directly applied. NgR1 was expressed in neural stem/progenitor cells (NSPs) derived from the adult spinal cord. Both an NgR1 antagonist NEP1-40 and ecto-domain phosphorylation of NgR1 promote neuronal cell production of the NSPs, in vitro. Thus, inhibition of NgR1 in NSPs can promote neuronal cell production, which could contribute to the enhanced recovery of locomotor function following infusion of PKA and ATP.

scholarly journals Retinoic acid-dependent attraction of adult spinal cord axons towards regenerating newt limb blastemas in vitro

2005 ◽  
Vol 281 (1) ◽  
pp. 112-120 ◽  
Author(s):  
Jennifer M. Dmetrichuk ◽  
Gaynor E. Spencer ◽  
Robert L. Carlone
Keyword(s):  
Spinal Cord ◽  
Retinoic Acid ◽  
Adult Spinal Cord ◽  
Newt Limb

scholarly journals Upregulation of Apol8 by Epothilone D facilitates the neuronal relay of transplanted NSCs in spinal cord injury

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Weiwei Xue ◽  
Haipeng Zhang ◽  
Yongheng Fan ◽  
Zhifeng Xiao ◽  
Yannan Zhao ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Neuronal Differentiation ◽  
Target Gene ◽  
Rna Seq ◽  
Lesion Site ◽  
Axonal Sprouting ◽  
Epothilone D ◽  
Cord Injury

Abstract Background Microtubule-stabilizing agents have been demonstrated to modulate axonal sprouting during neuronal disease. One such agent, Epothilone D, has been used to treat spinal cord injury (SCI) by promoting axonal sprouting at the lesion site after SCI. However, the role of Epothilone D in the differentiation of neural stem cells (NSCs) in SCI repair is unknown. In the present study, we mainly explored the effects and mechanisms of Epothilone D on the neuronal differentiation of NSCs and revealed a potential new SCI treatment. Methods In vitro differentiation assays, western blotting, and quantitative real-time polymerase chain reaction were used to detect the effects of Epothilone D on NSC differentiation. Retrograde tracing using a pseudotyped rabies virus was then used to detect neuronal circuit construction. RNA sequencing (RNA-Seq) was valuable for exploring the target gene involved in the neuronal differentiation stimulated by Epothilone D. In addition, lentivirus-induced overexpression and RNA interference technology were applied to demonstrate the function of the target gene. Last, an Apol8-NSC-linear ordered collagen scaffold (LOCS) graft was prepared to treat a mouse model of SCI, and functional and electrophysiological evaluations were performed. Results We first revealed that Epothilone D promoted the neuronal differentiation of cultured NSCs and facilitated neuronal relay formation in the injured site after SCI. Furthermore, the RNA-Seq results demonstrated that Apol8 was upregulated during Epothilone D-induced neuronal relay formation. Lentivirus-mediated Apol8 overexpression in NSCs (Apol8-NSCs) promoted NSC differentiation toward neurons, and an Apol8 interference assay showed that Apol8 had a role in promoting neuronal differentiation under the induction of Epothilone D. Last, Apol8-NSC transplantation with LOCS promoted the neuronal differentiation of transplanted NSCs in the lesion site as well as synapse formation, thus improving the motor function of mice with complete spinal cord transection. Conclusions Epothilone D can promote the neuronal differentiation of NSCs by upregulating Apol8, which may provide a promising therapeutic target for SCI repair.

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