Effects of delayed repair of peripheral nerve injury on the spatial distribution of motor endplates in target muscle

Motor endplates(MEPs) are important sites of information exchange between motor neurons and skeletal muscle, and are distributed in an organized pattern of lamellae in the muscle. Delayed repair of peripheral nerve injury typically results in unsatisfactory functional recovery because of MEP degeneration. In this study, the mouse tibial nerve was transected and repaired with a biodegradable chitin conduit, immediately following or 1 or 3 months after the injury. Fluorescent α-bungarotoxin was injected to label MEPs. Tissue optical clearing combined with light-sheet microscopy revealed that MEPs were distributed in an organized pattern of lamellae in skeletal muscle after delayed repair for 1 and 3 months. However, the total number of MEPs, the number of MEPs per lamellar cluster, and the maturation of single MEPs in gastrocnemius muscle gradually decreased with increasing denervation time. These findings suggest that delayed repair can restore the spatial distribution of MEPs, but it has an adverse effect on the homogeneity of MEPs in the lamellar clusters and the total number of MEPs in the target muscle. The study procedures were approved by the Animal Ethics Committee of the Peking University People's Hospital(approval No. 2019 PHC015) on April 8, 2019.

Stability of motor endplates is greater in the biceps than in the interossei in a rat model of obstetric brachial plexus palsy

The time window for repair of the lower trunk is shorter than that of the upper trunk in patients with obstetric brachial plexus palsy. The denervated intrinsic muscles of the hand become irreversibly atrophic much faster than the denervated biceps. However, it is unclear whether the motor endplates of the denervated interosseous muscles degenerate more rapidly than those of the denervated biceps. In this study, we used a rat model of obstetric brachial plexus palsy of the right upper limb. C5–6 was lacerated distal to the intervertebral foramina, with concurrent avulsion of C7–8 and T1, with the left upper limb used as the control. Bilateral interossei and biceps were collected at 5 and 7 weeks. Immunofluorescence was used to assess the morphology of the motor endplates. Real-time quantitative polymerase chain reaction and western blot assay were used to assess mRNA and protein expression levels of acetylcholine receptor subunits(α, β and δ), rapsyn and β-catenin. Immunofluorescence microscopy showed that motor endplates in the denervated interossei were fragmented, while those in the denervated biceps were morphologically intact with little fragmentation. The number and area of motor endplates, relative to the control side, were significantly lower in the denervated interossei compared with the denervated biceps. mRNA and protein expression levels of acetylcholine receptor subunits(α, β and δ) were significantly lower, whereas β-catenin protein expression was higher, in the denervated interossei compared with the denervated biceps. The protein expression of rapsyn was higher in the denervated biceps than in the denervated interossei at 7 weeks. Our findings demonstrate that motor endplates of interossei are destabilized, whereas those of the biceps remain stable, in the rat model of obstetric brachial plexus palsy. All procedures were approved by the Experimental Animal Ethics Committee of Fudan University, China(approval No. DF-187) in January 2016.

Basic fibroblast growth factor attenuates the degeneration of injured spinal cord motor endplates

The distal end of the spinal cord and neuromuscular junction may develop secondary degeneration and damage following spinal cord injury because of the loss of neural connections. In this study, a rat model of spinal cord injury, established using a modified Allen＇s method, was injected with basic fibroblast growth factor solution via subarachnoid catheter. After injection, rats with spinal cord injury displayed higher scores on the Basso, Beattie and Bresnahan locomotor scale. Motor function was also well recovered and hematoxylin-eosin staining showed that spinal glial scar hyperplasia was not apparent. Additionally, anterior tibial muscle fibers slowly, but progressively, atrophied. Immu- nohistochemical staining showed that the absorbance values of calcitonin gene related peptide and acetylcholinesterase in anterior tibial muscle and spinal cord were similar, and injection of basic fi- broblast growth factor increased this absorbance. Results showed that after spinal cord injury, the distal motor neurons and motor endplate degenerated. Changes in calcitonin gene related peptide and acetylcholinesterase in the spinal cord anterior horn motor neurons and motor endplate then occurred that were consistent with this regeneration. Our findings indicate that basic fibroblast growth factor can protect the endplate through gene related peptide and acetylcholinesterase cord. attenuating the decreased expression of calcitonin n anterior horn motor neurons of the injured spinal

The anatomical,electrophysiological and histological observations of muscle contraction units in rabbits:a new perspective on nerve injury and regeneration

In the conventional view a muscle is composed of intermediate structures before its further division into microscopic muscle fibers.Our experiments in mice have confirmed this intermediate structure is composed of the lamella cluster formed by motor endplates,the innervating nerve branches and the corresponding muscle fibers,which can be viewed as an independent structural and functional unit.In this study,we verified the presence of these muscle construction units in rabbits.The results showed that the muscular branch of the femoral nerve sent out 4–6 nerve branches into the quadriceps and the tibial nerve sent out 4–7 nerve branches into the gastrocnemius.When each nerve branch of the femoral nerve was stimulated from the most lateral to the medial,the contraction of the lateral muscle,intermediate muscle and medial muscle of the quadriceps could be induced by electrically stimulating at least one nerve branch.When stimulating each nerve branch of the tibial nerve from the lateral to the medial,the muscle contraction of the lateral muscle 1,lateral muscle 2,lateral muscle 3 and medial muscle of the gastrocnemius could be induced by electrically stimulating at least one nerve branch.Electrical stimulation of each nerve branch resulted in different electromyographical waves recorded in different muscle subgroups.Hematoxylin-eosin staining showed most of the nerve branches around the neuromuscular junctions consisted of one individual neural tract,a few consisted of two or more neural tracts.The muscles of the lower limb in the rabbit can be subdivided into different muscle subgroups,each innervated by different nerve branches,thereby allowing much more complex muscle activities than traditionally stated.Together,the nerve branches and the innervated muscle subgroups can be viewed as an independent structural and functional unit.This study was approved by the Animal Ethics Committee of Peking University People’s Hospital(approval No.2019 PHE027)on October 20,2019.