A"messenger zone hypothesis"based on the visual three-dimensional spatial distribution of motoneurons innervating deep limb muscles

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.

Peripheral nerve fibroblasts secrete neurotrophic factors to promote axon growth of motoneurons

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.

Time windows for postnatal changes in morphology and membrane excitability of genioglossal and oculomotor motoneurons

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.

BDNF secreted from nerve fibroblasts promotes neurite outgrowth of motoneurons by activating ERK and AKT pathway

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.

Expression of neurofilament gene in spinal motoneurons during neural regeneration

The technique in situ hybridization was used to measure the levels of light （NF-L）,medium（NF-M） and heavy （NF-H） neurofilament protein subunits mRNA in L4-6 spinal motoneurons in adult rat during regeneration following a unilateral crush of the sciatic nerve.It was found that the hybridization signals of each neurofilament subunit mRNA were dramatically decreased in spinal motoneurons postaxotomy by light microscopy. The hybridization signals of NF-L and NF-M mRNA were located in cytoplasm of neurons, whereas NF-H mRNA was found in both nucleus and cytoplasm of neurons. lmage analysis showed that the encoding levels of mRNA for each of neurofilament subunit mRNA reduced on the 3rd d and returned to control levels on the 28th d following the lesion. The relative levels of mRNA coding for each neurofilament subunit were significantly different. The lowest level of NF-L mRNA was observed at 5 d postaxotomy, and that of NF-M, NF-H mRNA on the 7th and 10th d after injury. Moreover,the levels of HF-M and

Expression of α Tubulin Gene in Motoneurons of Spinal Cord During Nerve Regeneration

We have previously reported that the injury of sciatic nerve causes not only the change of axonal ultrastructures, but also the increase of β tubulin gene expression in the neural soma. And the microtubule, as a carrier of transported materials, of axonal architecture plays an important role. It has also been found that the content and transported velocity of tubulin in the regeneration nerve increase significantly owing to the requirements of axonal reconstruction. As the NOR-Ag staining technique is specific

Downregulation of EphB2 by RNA interference attenuates glial/fibrotic scar formation and promotes axon growth

The rapid formation of a glial/fibrotic scar is one of the main factors hampering axon growth after spinal cord injury. The bidirectional Eph B2/ephrin-B2 signaling of the fibroblast-astrocyte contact-dependent interaction is a trigger for glial/fibrotic scar formation. In the present study, a new in vitro model was produced by coculture of fibroblasts and astrocytes wounded by scratching to mimic glial/fibrotic scar-like structures using an improved slide system. After treatment with RNAi to downregulate Eph B2, changes in glial/fibrotic scar formation and the growth of VSC4.1 motoneuron axons were examined. Following RNAi treatment, fibroblasts and astrocytes dispersed without forming a glial/fibrotic scar-like structure. Furthermore, the expression levels of neurocan, NG2 and collagen I in the coculture were reduced, and the growth of VSC4.1 motoneuron axons was enhanced. These findings suggest that suppression of Eph B2 expression by RNAi attenuates the formation of a glial/fibrotic scar and promotes axon growth. This study was approved by the Laboratory Animal Ethics Committee of Jiangsu Province, China(approval No. 2019-0506-002) on May 6, 2019.

Current landscape in motoneuron regeneration and reconstruction for motor cranial nerve injuries

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.

Neural computational modeling reveals a major role of corticospinal gating of central oscillations in the generation of essential tremor

Essential tremor, also referred to as familial tremor, is an autosomal dominant genetic disease and the most common movement disorder. It typically involves a postural and motor tremor of the hands, head or other part of the body. Essential tremor is driven by a central oscillation signal in the brain. However, the corticospinal mechanisms involved in the generation of essential tremor are unclear. Therefore, in this study, we used a neural computational model that includes both monosynaptic and multisynaptic corticospinal pathways interacting with a propriospinal neuronal network. A virtual arm model is driven by the central oscillation signal to simulate tremor activity behavior. Cortical descending commands are classified as alpha or gamma through monosynaptic or multisynaptic corticospinal pathways, which converge respectively on alpha or gamma motoneurons in the spinal cord. Several scenarios are evaluated based on the central oscillation signal passing down to the spinal motoneurons via each descending pathway. The simulated behaviors are compared with clinical essential tremor characteristics to identify the corticospinal pathways responsible for transmitting the central oscillation signal. A propriospinal neuron with strong cortical inhibition performs a gating function in the generation of essential tremor. Our results indicate that the propriospinal neuronal network is essential for relaying the central oscillation signal and the production of essential tremor.

Network-centric medicine for peripheral nerve injury:treating the whole to boost endogenous mechanisms of neuroprotection and regeneration

Peripheral nerve injuries caused by accidents may lead to paralysis,sensory disturbances,anaesthesia,and lack of autonomic functions.Functional recovery after disconnection of the motoneuronal soma from target tissue with proximal rupture of axons is determined by several factors:motoneuronal soma viability,proper axonal sprouting across inhibitory zones and elongation toward specific muscle,effective synapse contact rebuilding,and prevention of muscle atrophy.Therapies,such as adjuvant drugs with pleiotropic effects,that promote functional recovery after peripheral nerve injury are needed.Toward this aim,we designed a drug discovery workflow based on a network-centric molecular vision using unbiased proteomic data and neural artificial computational tools.Our focus is on boosting intrinsic capabilities of neurons for neuroprotection;this is in contrast to the common approach based on suppression of a pathobiological pathway known to be associated with disease condition.Using our workflow,we discovered neuroheal,a combination of two repurposed drugs that promotes motoneuronal soma neuroprotection,is anti-inflammatory,enhances axonal regeneration after axotomy,and reduces muscle atrophy.This drug discovery workflow has thus yielded a therapy that is close to its clinical application.

NEUROBIOLOGICAL EFFECTS OF NRCF DERIVED FROM DISTAL STUMPS OF MOTOR NERVE AND SENSORY NERVE AND b-FGF ON CULTURED MOTONEURON IN VITRO

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.

神经递质释放中的有关蛋白

神经递质释放中的有关蛋白ＮＩＰＳ１９９５年２月号综述了神经递质释放过程中的许多重要蛋白质的作用。这些蛋白质分属于突触囊泡膜、胞液及神经末梢质膜。在突触囊泡膜上的蛋白有：Ｓｙｎａｐｔｏｂｒｅｖｉｎ，Ｓｙｎａｐｔｏｔａｇｍｉｎ，Ｓｙｎａｐｔｏｐｈｙ－ｓｉ...

EFFECT OF 22kD,35kD PROTEIN MOLECULES FROM EXTRACT OF SKELETAL MUSCLE ON CULTURED ANTERIOR HORN MOTONEURON OF LUMBAR SPINE IN RAT

Ⅰ. INTRODUCTION The survival of motoneurons in vertebrates during their embryonic period depends on certain neurotrophic substances provided from their skeletal muscles. Skeletal muscles have been shown, in in vitro and in vivo experiments, to contain substances which are capable of enhancing the survival and development of motoneurons but preventing the

Relationship between the partitioning properties of motoneuron recruitment and the needle feeling propagation

Histological and physiological studies indicated that most skeletal muscles can be divided into a series of relatively independent sub-volumes: "neuromuscular compartments" and the partitioning property of the muscle result in the localization of muscle reflex. In the present experiment we studied the recruitment properties of medial gastracnemius (MG) muscle motoneuron pool of decerebrate cat with two kinds of local mechanical stimulation: local stretch and acupuncture-like stimuli. The results indicate that: (ⅰ) there is an obvious property of localization of recruitment activity, only the MNs which innervate the stimulated compartment were recruited by weaker and shorter stimuli; (ⅱ) recruitment activity spread to those MNs which supply the adjacent and distal compartments during the strength of stimulation or duration of the stimulation was increased; and (ⅲ) the recruitment property of muscle activity elicited by the local mechanical stimulation is thought similar to that of "needle feeling" along the meridian pathway during stimulation of acupuncture point.