Coordinated contraction of skeletal muscles relies on selective connections between the muscles and multiple classes of the spinal motoneuro ns.Howeve r,current research on the spatial location of the spinal motoneuro...Coordinated contraction of skeletal muscles relies on selective connections between the muscles and multiple classes of the spinal motoneuro ns.Howeve r,current research on the spatial location of the spinal motoneurons innervating differe nt muscles is limited.In this study,we investigated the spatial distribution and relative position of different motoneurons that control the deep muscles of the mouse hindlimbs,which were innervated by the obturator nerve,femoral nerve,inferior gluteal nerve,deep pe roneal nerve,and tibial nerve.Locations were visualized by combining a multiplex retrograde tracking technique compatible with three-dimensional imaging of solvent-cleared o rgans(3DISCO)and 3-D imaging technology based on lightsheet fluorescence microscopy(LSFM).Additionally,we propose the hypothesis that"messenger zones"exist as interlaced areas between the motoneuron pools that dominate the synergistic or antagonist muscle groups.We hypothesize that these interlaced neurons may participate in muscle coordination as messenger neurons.Analysis revealed the precise mutual positional relationships among the many motoneurons that innervate different deep muscles of the mouse.Not only do these findings update and supplement our knowledge regarding the overall spatial layout of spinal motoneurons that control mouse limb muscles,but they also provide insights into the mechanisms through which muscle activity is coordinated and the architecture of motor circuits.展开更多
目的探究踝关节跖背屈疲劳任务中高精度经颅直流电刺激(high-definition transcranial direct current stimulation,HD-tDCS)对H-反射和M-波的调控效果,为HD-tDCS减轻神经肌肉疲劳的实际应用提供方向。方法招募20名健康青年男性受试者,...目的探究踝关节跖背屈疲劳任务中高精度经颅直流电刺激(high-definition transcranial direct current stimulation,HD-tDCS)对H-反射和M-波的调控效果,为HD-tDCS减轻神经肌肉疲劳的实际应用提供方向。方法招募20名健康青年男性受试者,随机分为真刺激组和假刺激组各10名。对受试者采取连续5 d的单盲HD-tDCS干预(时间20 min;强度2 mA;靶点Cz),干预前1天采集受试者安静条件下的H-反射和M-波,跖屈肌最大自主收缩(maximal voluntary isometric contraction,MVIC)时的M-波,跖屈肌和背屈肌MVIC力矩,并进行一次踝关节跖背屈运动性疲劳任务,以确定受试者达到该任务疲劳的时间。干预后1天进行与第1次疲劳任务相同的运动时间,并进行后测的数据采集。采用重复测量双因素(刺激方案×疲劳前后)方差分析其自变量对受试者肌肉力学特性、α运动神经元传导特性各指标的影响。结果相较于疲劳前,两组疲劳后的自主激活值(voluntary activation,VA)、H-反射最大值(maximal H-reflex,H_(max))、M-波最大值(maximal M-wave,Mmax)、跖屈肌和背屈肌MVIC力矩均显著降低(P<0.05),但相比于真刺激组,假刺激组的VA和背屈肌MVIC力矩下降更为显著(P<0.05)。结论连续5 d的HD-tDCS干预有助于提高脊髓节段α运动神经元的活性,且能抑制跖背屈疲劳诱发的外周“神经-肌肉”接头处信息传递能力的下降。展开更多
The intricate anatomy and physiology of cranial nerves have inspired clinicians and scientists to study their roles in the nervous system. Damage to motor cranial nerves may result from a variety of organic or iatroge...The intricate anatomy and physiology of cranial nerves have inspired clinicians and scientists to study their roles in the nervous system. Damage to motor cranial nerves may result from a variety of organic or iatrogenic insults and causes devastating functional impairment and disfigurement. Surgical innovations directed towards restoring function to injured motor cranial nerves and their associated organs have evolved to include nerve repair, grafting, substitution, and muscle transposition. In parallel with this progress, research on tissue-engineered constructs, development of bioelectrical interfaces, and modulation of the regenerative milieu through cellular, immunomodulatory, or neurotrophic mechanisms has proliferated to enhance the available repertoire of clinically applicable reconstructive options. Despite these advances, patients continue to suffer from functional limitations relating to inadequate cranial nerve regeneration, aberrant reinnervation, or incomplete recovery of neuromuscular function. These shortfalls have profound quality of life ramifications and provide an impetus to further elucidate mechanisms underlying cranial nerve denervation and to improve repair. In this review, we summarize the literature on reconstruction and regeneration of motor cranial nerves following various injury patterns. We focus on seven cranial nerves with predominantly efferent functions and highlight shared patterns of injuries and clinical manifestations. We also present an overview of the existing reconstructive approaches, from facial reanimation, laryngeal reinnervation, to variations of interposition nerve grafts for reconstruction. We discuss ongoing endeavors to promote nerve regeneration and to suppress aberrant reinnervation and the development of synkinesis. Insights from these studies will shed light on recent progress and new horizons in understanding the biomechanics of peripheral nerve neurobiology, with emphasis on promising strategies for optimizing neural regeneration and identifying future directions in the field of motor cranial neuron research.展开更多
Our previous study revealed that early application of electrical field stimulation（EFS） with the anode at the lesion and the cathode distal to the lesion reduced injury potential, inhibited secondary injury and was ...Our previous study revealed that early application of electrical field stimulation（EFS） with the anode at the lesion and the cathode distal to the lesion reduced injury potential, inhibited secondary injury and was neuroprotective in the dorsal corticospinal tract after spinal cord injury（SCI）. The objective of this study was to further evaluate the effect of EFS on protection of anterior horn motoneurons and their target musculature after SCI and its mechanism. Rats were randomized into three equal groups. The EFS group received EFS for 30 minutes immediately after injury at T_（10）. SCI group rats were only subjected to SCI and sham group rats were only subjected to laminectomy. Luxol fast blue staining demonstrated that spinal cord tissue in the injury center was better protected; cross-sectional area and perimeter of injured tissue were significantly smaller in the EFS group than in the SCI group. Immunofluorescence and transmission electron microscopy showed that the number of spinal cord anterior horn motoneurons was greater and the number of abnormal neurons reduced in the EFS group compared with the SCI group. Wet weight and cross-sectional area of vastus lateralis muscles were smaller in the SCI group to in the sham group. However, EFS improved muscle atrophy and behavioral examination showed that EFS significantly increased the angle in the inclined plane test and Tarlov＇s motor grading score. The above results confirm that early EFS can effectively impede spinal cord anterior horn motoneuron loss, promote motor function recovery and reduce muscle atrophy in rats after SCI.展开更多
BZ]Dendro dendritic and dendro somatic projections are common between spinal motoneurons. We attempted to clarify whether there are functional connections through these projections. Methods. Motoneurons were antidromi...BZ]Dendro dendritic and dendro somatic projections are common between spinal motoneurons. We attempted to clarify whether there are functional connections through these projections. Methods. Motoneurons were antidromically stimulated by the muscle nerve and recorded intracellularly to examine the direct interaction between them, after the related dorsal roots had been cut. Results. Excitatory connections, demonstrated by depolarizing potentials in response to muscle nerve stimulation, were found between motoneurons innervating the same muscle or synergistic muscles, but never between motoneurons innervating antagonistic muscles. These potentials were finely graded in response to a series of increasing stimuli and resistant to high frequency (50Hz) stimulation. Conclusions.These results indicate that excitatory connections, with certain specificity of spatial and temporal distribution, occur in the spinal motoneurons. It is also suggested that electrical coupling should be involved in these connections and this mechanism should improve the excitability of the motoneurons in the same column.展开更多
To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed ra...To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed rat models of spinal cord contusion injury, 7 days later, injected adipose-derived stem cells-transdifferentiated motoneurons into the epicenter, rostral and caudal regions of the impact site and simultaneously transplanted glial cell line-derived neuro-trophic factor-gelfoam complex into the myelin sheath. Motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery reduced cavity formations and increased cell density in the transplantation site. The combined therapy exhibited superior promoting effects on recovery of motor function to transplantation of glial cell line-derived neurotrophic factor, adipose-derived stem cells or motoneurons alone. These ifndings suggest that motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery holds a great promise for repair of spinal cord injury.展开更多
Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different p...Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different phenotypes can guide the migration of Schwann cells to the same sensory or motor phenotype.In this study,we analyzed the different effects of peripheral nerve-derived fibroblasts and cardiac fibroblasts on motoneurons.Compared with cardiac fibroblasts,peripheral nerve fibroblasts greatly promoted motoneuron neurite outgrowth.Transcriptome analysis results identified 491 genes that were differentially expressed in peripheral nerve fibroblasts and cardiac fibroblasts.Among these,130 were significantly upregulated in peripheral nerve fibroblasts compared with cardiac fibroblasts.These genes may be involved in axon guidance and neuron projection.Three days after sciatic nerve transection in rats,peripheral nerve fibroblasts accumulated in the proximal and distal nerve stumps,and most expressed brain-derived neurotrophic factor.In vitro,brain-derived neurotrophic factor secreted from peripheral nerve fibroblasts increased the expression ofβ-actin and F-actin through the extracellular regulated protein kinase and serine/threonine kinase pathways,and enhanced motoneuron neurite outgrowth.These findings suggest that peripheral nerve fibroblasts and cardiac fibroblasts exhibit different patterns of gene expression.Peripheral nerve fibroblasts can promote motoneuron neurite outgrowth.展开更多
Objective: To explore the mechanism of neurotropism in peripheral nerve regeneration by assessing the bioactivity of regeneration microcircumstance on motoneurons. Methods The motor branch Of femoral nerve to quadrice...Objective: To explore the mechanism of neurotropism in peripheral nerve regeneration by assessing the bioactivity of regeneration microcircumstance on motoneurons. Methods The motor branch Of femoral nerve to quadriceps was incised and the distal stump was sutured with one-end silicone chamber. The nerve regeneration chamber fluids from distal motor nerve stumps (motor branch of femoral nerve ) (MD-NRCF) was collected 7d post-operatively, and with the same method, nerve regeneration conditioned fluids from distal stumps nerve stumps (saphenous nerve ) (SD-NRCF) was collected. The dissociated rat’s motoneurons were co-cultured with MD-NRCF, SD-NRCF, basic fibroblast growth factor (b-FGF) and serum-free medium for 72h respectively and then were photographed under phase-contrast microscope. The longest neurites and cellbody areas of motoneurons from each group were measured by cell image processing computer system. MTT colorimetric assay was also used to measure cell activation. Results The cells of MD-NRCF group had significantly longer neurites than the other 3 groups, and their activation was also superior to those of the other groups. Conclusion These results indicate that MD-NRCF has more significant neurite-promoting and neurobiological effects on motoneuron than SD-NRCF and b-FGF.展开更多
BACKGROUND： Erythropoietin and recombinant human erythropoietin （rhEPO） inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO ...BACKGROUND： Erythropoietin and recombinant human erythropoietin （rhEPO） inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO can protect facial motoneurons （FMNs） as Well. OBJECTIVE： To test the neuroprotective effects of rhEPO on injured VMNs, as well as the influence on Caspase-3 expression. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. This study was performed at the Central Laboratory of Basic Medical College, Chongqing Medical University from January to October 2007. MATERIALS： Seventy-five female SD rats, weighing 210-230 g. rhEPO injection was provided by Sansheng pharmaceuticals company, Shenyang City, Liaoning Province, China, and the License number was HMLN S20010001. METHODS： A total of 75 female rats were randomly divided into rhEPO treatment, control, and sham operation groups, with 25 rats in each group. Rat models of facial nerve injury were established in the rhEPO treatment group and the control group by crushing the main trunk of the left facial nerve. Surgical microscopic observation of the facial nerve damage displayed perineurial disruption. The left stylomastoid foramen of the sham operation group were only exposed, but without nerve injury. The rhEPO treatment group was treated with rhEPO （5 000 U/kg, i.p.） once following injury and once a day for two weeks. The control and sham operation groups were treated with the same dose of normal saline （i.p.）, once following injury and once a day for two weeks. MAIN OUTCOME MEASURES： Rats were sacrificed 3, 7, 14, 21, and 28 days after injury, FMN survival after facial nerve injury was analyzed by Toluidine blue staining, and then survival ratios （L/R） were calculated. The number of apoptotic profiles in the injured FMNs were evaluated by TUNEL staining. Expression of Caspase-3 in the facial nucleus was detected by immunohistochemistry methods. RESULTS： A total of 75 rats were included in the final analysis. FMN survival ratios, both in rhEPO treatment group and control group, decreased gradually between seven and 28 days; however, FMN survival ratios were significantly greater in the rhEPO treatment group compared to the control group （P 〈 0.05）. No TUNEL-positive cells were observed three days after injury in the rhEPO treatment and control groups; however, by seven days after injury, apoptotic cells were observed and peaked by 14 days in the control group. Between seven and 21 days, apoptotic cell numbers were significantly lower in the rhEPO treatment group compared to the control group （P 〈 0.05）. The expression of Caspase-3 increased three days after injury and peaked at 14 days in the control group. Nevertheless, Caspase-3 expression was significantly lower in the rhEPO treatment group compared to the control group at each time point （P 〈 0.05）. CONCLUSION： Treatment with rhEPO can effectively protect facial motoneurons by reducing expression of Caspase-3 and inhibiting apoptosis.展开更多
The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by compari...The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by comparing immunoreactivity for p75 and phosphorylated c-Jun （p-c-Jun）, the production of JNK activation in axotomized motoneurons in postnatal day （PN）I, PN7, PN14 and adult rats. Intensive p-c-Jun was induced in axotomized motoneurons in PN1 and PN7. In PN14, p-c-Jun expression was sharply reduced after the same injury. The decreased expression of p-c-Jun at this age coincided with a developmental switch of re-expression of p75 in axotomized cells. In adult animals, no p-c-Jun but intensive p75 was detected in axotomized motoneurons. These results indicate differential expression or turnover of phosphorylation of c-Jun and p75 in immature versus mature spinal motoneurons in response to axonal injury. The non-co-occurrence of p75 and p-c-Jun in injured motoneurons indicated that p75 may not activate JNK pathway, suggesting that the p75 may not be involved in cell death in axotomized motoneurons.展开更多
Time windows for postnatal changes in morphology and membrane excitability of genioglossal(GG) and oculomotor(OCM) motoneurons(MNs) are yet to be fully described. Analysis of data on brain slices in vitro of the 2 pop...Time windows for postnatal changes in morphology and membrane excitability of genioglossal(GG) and oculomotor(OCM) motoneurons(MNs) are yet to be fully described. Analysis of data on brain slices in vitro of the 2 populations of MNs point to a well-defined developmental program that progresses with common age-related changes characterized by:(1) increase of dendritic surface along with length and reshaping of dendritic tree complexity;(2) disappearance of gap junctions early in development;(3) decrease of membrane passive properties, such as input resistance and time constant, together with an increase in the number of cells displaying sag, and modifications in rheobase;(4) action potential shortening and afterhyperpolarization; and(5) an increase in gain and maximum firing frequency. These modifications take place at different time windows for each motoneuronal population. In GG MNs, active membrane properties change mainly during the first postnatal week, passive membrane properties in the second week, and dendritic increasing length and size in the third week of development. In OCM MNs, changes in passive membrane properties and growth of dendritic size take place during the first postnatal week, while active membrane properties and rheobase change during the second and third weeks of development. The sequential order of changes is inverted between active and passive membrane properties, and growth in size does not temporally coincide for both motoneuron populations. These findings are discussed on the basis of environmental cues related to maturation of the respiratory and OCM systems.展开更多
Peripheral nerve fibroblasts play an important role in the process of nerve development and regeneration.The present study revealed that fibroblasts with different tissue sources,such as peripheral nerve fibroblasts(N...Peripheral nerve fibroblasts play an important role in the process of nerve development and regeneration.The present study revealed that fibroblasts with different tissue sources,such as peripheral nerve fibroblasts(N-Fbs)and cardiac Fbs(C-Fbs),exerted distinct effects on motoneurons.Compared to C-Fbs,N-Fbs significantly promoted neurite outgrowth of motoneurons in vitro.mRNA sequencing identified a total of 491 differentially expressed genes between N-Fbs and C-Fbs.Out of them,130 genes were significantly upregulated in N-Fbs than in C-Fbs,and these genes might be involved in axon guidance and neuron projection.展开更多
Vascular endothelial growth factor（VEGF）,an angiogenic factor with neuroprotective effects：The VEGF was initially characterized by its vasculogenic and angiogenic activities and its capacity to promote vascular per...Vascular endothelial growth factor（VEGF）,an angiogenic factor with neuroprotective effects：The VEGF was initially characterized by its vasculogenic and angiogenic activities and its capacity to promote vascular permeability（Yancopoulos et al.,2000）.VEGF is also known as VEGF-A and is the prototype member of a related group of five trophic factors,VEGF-B,VEGF-C,VEGF-D and placental growth factor（Pl GF;Lange et al.,2016）.展开更多
基金supported by the Chinese National General Program of the National Natural Science Foundation of China,No.82072162(to XY)。
文摘Coordinated contraction of skeletal muscles relies on selective connections between the muscles and multiple classes of the spinal motoneuro ns.Howeve r,current research on the spatial location of the spinal motoneurons innervating differe nt muscles is limited.In this study,we investigated the spatial distribution and relative position of different motoneurons that control the deep muscles of the mouse hindlimbs,which were innervated by the obturator nerve,femoral nerve,inferior gluteal nerve,deep pe roneal nerve,and tibial nerve.Locations were visualized by combining a multiplex retrograde tracking technique compatible with three-dimensional imaging of solvent-cleared o rgans(3DISCO)and 3-D imaging technology based on lightsheet fluorescence microscopy(LSFM).Additionally,we propose the hypothesis that"messenger zones"exist as interlaced areas between the motoneuron pools that dominate the synergistic or antagonist muscle groups.We hypothesize that these interlaced neurons may participate in muscle coordination as messenger neurons.Analysis revealed the precise mutual positional relationships among the many motoneurons that innervate different deep muscles of the mouse.Not only do these findings update and supplement our knowledge regarding the overall spatial layout of spinal motoneurons that control mouse limb muscles,but they also provide insights into the mechanisms through which muscle activity is coordinated and the architecture of motor circuits.
基金supported by the United States National Institute of Health grant 1K08DC012535(to MJB)Program for Neurology Research and Discoverythe Sinai Medical Staff Foundation Neuroscience Scholar Fund(to ELF)
文摘The intricate anatomy and physiology of cranial nerves have inspired clinicians and scientists to study their roles in the nervous system. Damage to motor cranial nerves may result from a variety of organic or iatrogenic insults and causes devastating functional impairment and disfigurement. Surgical innovations directed towards restoring function to injured motor cranial nerves and their associated organs have evolved to include nerve repair, grafting, substitution, and muscle transposition. In parallel with this progress, research on tissue-engineered constructs, development of bioelectrical interfaces, and modulation of the regenerative milieu through cellular, immunomodulatory, or neurotrophic mechanisms has proliferated to enhance the available repertoire of clinically applicable reconstructive options. Despite these advances, patients continue to suffer from functional limitations relating to inadequate cranial nerve regeneration, aberrant reinnervation, or incomplete recovery of neuromuscular function. These shortfalls have profound quality of life ramifications and provide an impetus to further elucidate mechanisms underlying cranial nerve denervation and to improve repair. In this review, we summarize the literature on reconstruction and regeneration of motor cranial nerves following various injury patterns. We focus on seven cranial nerves with predominantly efferent functions and highlight shared patterns of injuries and clinical manifestations. We also present an overview of the existing reconstructive approaches, from facial reanimation, laryngeal reinnervation, to variations of interposition nerve grafts for reconstruction. We discuss ongoing endeavors to promote nerve regeneration and to suppress aberrant reinnervation and the development of synkinesis. Insights from these studies will shed light on recent progress and new horizons in understanding the biomechanics of peripheral nerve neurobiology, with emphasis on promising strategies for optimizing neural regeneration and identifying future directions in the field of motor cranial neuron research.
基金supported by the National Natural Science Foundation of China,No.31400717,51577183the Natural Science Foundation of Beijing of China,No.7164317the Youth Innovation Promotion Association CAS,No.2018172
文摘Our previous study revealed that early application of electrical field stimulation（EFS） with the anode at the lesion and the cathode distal to the lesion reduced injury potential, inhibited secondary injury and was neuroprotective in the dorsal corticospinal tract after spinal cord injury（SCI）. The objective of this study was to further evaluate the effect of EFS on protection of anterior horn motoneurons and their target musculature after SCI and its mechanism. Rats were randomized into three equal groups. The EFS group received EFS for 30 minutes immediately after injury at T_（10）. SCI group rats were only subjected to SCI and sham group rats were only subjected to laminectomy. Luxol fast blue staining demonstrated that spinal cord tissue in the injury center was better protected; cross-sectional area and perimeter of injured tissue were significantly smaller in the EFS group than in the SCI group. Immunofluorescence and transmission electron microscopy showed that the number of spinal cord anterior horn motoneurons was greater and the number of abnormal neurons reduced in the EFS group compared with the SCI group. Wet weight and cross-sectional area of vastus lateralis muscles were smaller in the SCI group to in the sham group. However, EFS improved muscle atrophy and behavioral examination showed that EFS significantly increased the angle in the inclined plane test and Tarlov＇s motor grading score. The above results confirm that early EFS can effectively impede spinal cord anterior horn motoneuron loss, promote motor function recovery and reduce muscle atrophy in rats after SCI.
基金This study was supported by the grant from ClimbingProgram of Chinese Committee of Science
文摘BZ]Dendro dendritic and dendro somatic projections are common between spinal motoneurons. We attempted to clarify whether there are functional connections through these projections. Methods. Motoneurons were antidromically stimulated by the muscle nerve and recorded intracellularly to examine the direct interaction between them, after the related dorsal roots had been cut. Results. Excitatory connections, demonstrated by depolarizing potentials in response to muscle nerve stimulation, were found between motoneurons innervating the same muscle or synergistic muscles, but never between motoneurons innervating antagonistic muscles. These potentials were finely graded in response to a series of increasing stimuli and resistant to high frequency (50Hz) stimulation. Conclusions.These results indicate that excitatory connections, with certain specificity of spatial and temporal distribution, occur in the spinal motoneurons. It is also suggested that electrical coupling should be involved in these connections and this mechanism should improve the excitability of the motoneurons in the same column.
基金funded by Shefa Neurosciences Research Center at Khatam Al-Anbia Hospital,Tehran,Iran(Grant#86-N-105)
文摘To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed rat models of spinal cord contusion injury, 7 days later, injected adipose-derived stem cells-transdifferentiated motoneurons into the epicenter, rostral and caudal regions of the impact site and simultaneously transplanted glial cell line-derived neuro-trophic factor-gelfoam complex into the myelin sheath. Motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery reduced cavity formations and increased cell density in the transplantation site. The combined therapy exhibited superior promoting effects on recovery of motor function to transplantation of glial cell line-derived neurotrophic factor, adipose-derived stem cells or motoneurons alone. These ifndings suggest that motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery holds a great promise for repair of spinal cord injury.
基金supported by the National Key Research and Development Program of China,No.2017YFA0104703(to FD)the National Natural Science Foundation of China(Major Program),No.92068112(to FD)+2 种基金Science and Technology Program of Nantong of China,No.JC2020035(to QRH)National Natural Science Foundation of China,Nos.31500927(to QRH)and 31870977(to HYS)the Priority Academic Program Development of Jiangsu High Education Institutions(PAPD)(to FD).
文摘Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different phenotypes can guide the migration of Schwann cells to the same sensory or motor phenotype.In this study,we analyzed the different effects of peripheral nerve-derived fibroblasts and cardiac fibroblasts on motoneurons.Compared with cardiac fibroblasts,peripheral nerve fibroblasts greatly promoted motoneuron neurite outgrowth.Transcriptome analysis results identified 491 genes that were differentially expressed in peripheral nerve fibroblasts and cardiac fibroblasts.Among these,130 were significantly upregulated in peripheral nerve fibroblasts compared with cardiac fibroblasts.These genes may be involved in axon guidance and neuron projection.Three days after sciatic nerve transection in rats,peripheral nerve fibroblasts accumulated in the proximal and distal nerve stumps,and most expressed brain-derived neurotrophic factor.In vitro,brain-derived neurotrophic factor secreted from peripheral nerve fibroblasts increased the expression ofβ-actin and F-actin through the extracellular regulated protein kinase and serine/threonine kinase pathways,and enhanced motoneuron neurite outgrowth.These findings suggest that peripheral nerve fibroblasts and cardiac fibroblasts exhibit different patterns of gene expression.Peripheral nerve fibroblasts can promote motoneuron neurite outgrowth.
文摘Objective: To explore the mechanism of neurotropism in peripheral nerve regeneration by assessing the bioactivity of regeneration microcircumstance on motoneurons. Methods The motor branch Of femoral nerve to quadriceps was incised and the distal stump was sutured with one-end silicone chamber. The nerve regeneration chamber fluids from distal motor nerve stumps (motor branch of femoral nerve ) (MD-NRCF) was collected 7d post-operatively, and with the same method, nerve regeneration conditioned fluids from distal stumps nerve stumps (saphenous nerve ) (SD-NRCF) was collected. The dissociated rat’s motoneurons were co-cultured with MD-NRCF, SD-NRCF, basic fibroblast growth factor (b-FGF) and serum-free medium for 72h respectively and then were photographed under phase-contrast microscope. The longest neurites and cellbody areas of motoneurons from each group were measured by cell image processing computer system. MTT colorimetric assay was also used to measure cell activation. Results The cells of MD-NRCF group had significantly longer neurites than the other 3 groups, and their activation was also superior to those of the other groups. Conclusion These results indicate that MD-NRCF has more significant neurite-promoting and neurobiological effects on motoneuron than SD-NRCF and b-FGF.
文摘BACKGROUND： Erythropoietin and recombinant human erythropoietin （rhEPO） inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO can protect facial motoneurons （FMNs） as Well. OBJECTIVE： To test the neuroprotective effects of rhEPO on injured VMNs, as well as the influence on Caspase-3 expression. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. This study was performed at the Central Laboratory of Basic Medical College, Chongqing Medical University from January to October 2007. MATERIALS： Seventy-five female SD rats, weighing 210-230 g. rhEPO injection was provided by Sansheng pharmaceuticals company, Shenyang City, Liaoning Province, China, and the License number was HMLN S20010001. METHODS： A total of 75 female rats were randomly divided into rhEPO treatment, control, and sham operation groups, with 25 rats in each group. Rat models of facial nerve injury were established in the rhEPO treatment group and the control group by crushing the main trunk of the left facial nerve. Surgical microscopic observation of the facial nerve damage displayed perineurial disruption. The left stylomastoid foramen of the sham operation group were only exposed, but without nerve injury. The rhEPO treatment group was treated with rhEPO （5 000 U/kg, i.p.） once following injury and once a day for two weeks. The control and sham operation groups were treated with the same dose of normal saline （i.p.）, once following injury and once a day for two weeks. MAIN OUTCOME MEASURES： Rats were sacrificed 3, 7, 14, 21, and 28 days after injury, FMN survival after facial nerve injury was analyzed by Toluidine blue staining, and then survival ratios （L/R） were calculated. The number of apoptotic profiles in the injured FMNs were evaluated by TUNEL staining. Expression of Caspase-3 in the facial nucleus was detected by immunohistochemistry methods. RESULTS： A total of 75 rats were included in the final analysis. FMN survival ratios, both in rhEPO treatment group and control group, decreased gradually between seven and 28 days; however, FMN survival ratios were significantly greater in the rhEPO treatment group compared to the control group （P 〈 0.05）. No TUNEL-positive cells were observed three days after injury in the rhEPO treatment and control groups; however, by seven days after injury, apoptotic cells were observed and peaked by 14 days in the control group. Between seven and 21 days, apoptotic cell numbers were significantly lower in the rhEPO treatment group compared to the control group （P 〈 0.05）. The expression of Caspase-3 increased three days after injury and peaked at 14 days in the control group. Nevertheless, Caspase-3 expression was significantly lower in the rhEPO treatment group compared to the control group at each time point （P 〈 0.05）. CONCLUSION： Treatment with rhEPO can effectively protect facial motoneurons by reducing expression of Caspase-3 and inhibiting apoptosis.
基金supported by Direct Grant (Project No.2030392) of the Chinese University of Hong KongHK Spinal Cord Injury FoundationNational Key Basic Research Support Foundation (973 Project: 2011CB504402)
文摘The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by comparing immunoreactivity for p75 and phosphorylated c-Jun （p-c-Jun）, the production of JNK activation in axotomized motoneurons in postnatal day （PN）I, PN7, PN14 and adult rats. Intensive p-c-Jun was induced in axotomized motoneurons in PN1 and PN7. In PN14, p-c-Jun expression was sharply reduced after the same injury. The decreased expression of p-c-Jun at this age coincided with a developmental switch of re-expression of p75 in axotomized cells. In adult animals, no p-c-Jun but intensive p75 was detected in axotomized motoneurons. These results indicate differential expression or turnover of phosphorylation of c-Jun and p75 in immature versus mature spinal motoneurons in response to axonal injury. The non-co-occurrence of p75 and p-c-Jun in injured motoneurons indicated that p75 may not activate JNK pathway, suggesting that the p75 may not be involved in cell death in axotomized motoneurons.
文摘Time windows for postnatal changes in morphology and membrane excitability of genioglossal(GG) and oculomotor(OCM) motoneurons(MNs) are yet to be fully described. Analysis of data on brain slices in vitro of the 2 populations of MNs point to a well-defined developmental program that progresses with common age-related changes characterized by:(1) increase of dendritic surface along with length and reshaping of dendritic tree complexity;(2) disappearance of gap junctions early in development;(3) decrease of membrane passive properties, such as input resistance and time constant, together with an increase in the number of cells displaying sag, and modifications in rheobase;(4) action potential shortening and afterhyperpolarization; and(5) an increase in gain and maximum firing frequency. These modifications take place at different time windows for each motoneuronal population. In GG MNs, active membrane properties change mainly during the first postnatal week, passive membrane properties in the second week, and dendritic increasing length and size in the third week of development. In OCM MNs, changes in passive membrane properties and growth of dendritic size take place during the first postnatal week, while active membrane properties and rheobase change during the second and third weeks of development. The sequential order of changes is inverted between active and passive membrane properties, and growth in size does not temporally coincide for both motoneuron populations. These findings are discussed on the basis of environmental cues related to maturation of the respiratory and OCM systems.
文摘Peripheral nerve fibroblasts play an important role in the process of nerve development and regeneration.The present study revealed that fibroblasts with different tissue sources,such as peripheral nerve fibroblasts(N-Fbs)and cardiac Fbs(C-Fbs),exerted distinct effects on motoneurons.Compared to C-Fbs,N-Fbs significantly promoted neurite outgrowth of motoneurons in vitro.mRNA sequencing identified a total of 491 differentially expressed genes between N-Fbs and C-Fbs.Out of them,130 genes were significantly upregulated in N-Fbs than in C-Fbs,and these genes might be involved in axon guidance and neuron projection.
基金supported by Ministerio de Economía y Competitividad-FEDER(Grant reference:BFU2015-64515-P) in Spaina scholar of MEC(BES-2016-077912) in Spain
文摘Vascular endothelial growth factor（VEGF）,an angiogenic factor with neuroprotective effects：The VEGF was initially characterized by its vasculogenic and angiogenic activities and its capacity to promote vascular permeability（Yancopoulos et al.,2000）.VEGF is also known as VEGF-A and is the prototype member of a related group of five trophic factors,VEGF-B,VEGF-C,VEGF-D and placental growth factor（Pl GF;Lange et al.,2016）.
