UNC5B Overexpression Alleviates Peripheral Neuropathic Pain by Stimulating Netrin-1-Dependent Autophagic Flux in Schwann Cells.

Lesions or diseases of the somatosensory system can cause neuropathic pain (NP). Schwann cell (SC) autophagy plays an important role in NP. Uncoordinated gene 5 homolog B (UNC5B), the canonical dependent receptor of netrin-1, is known to be exclusively expressed in SCs and involved in NP; however, the underlying mechanisms were unclear. A rat model of sciatic nerve chronic constriction injury (CCI) was used to induce peripheral neuropathic pain. Adeno-associated virus (AAV) overexpressing UNC5B was applied to the injured nerve, and an autophagy inhibitor, 3-mechyladenine (3-MA), was intraperitoneally injected in some animals. Behavioral tests were performed to evaluate NP, the morphology of the injured nerves was analyzed, and autophagy-related proteins were detected. A rat SC line (RSC96) undergoing oxygen and glucose deprivation (OGD) was used to mimic an ischemic setting to examine the role of UNC5B in autophagy. Local UNC5B overexpression alleviated CCI-induced NP and rescued myelin degeneration. Meanwhile, UNC5B overexpression improved CCI-induced impairment of autophagic flux, while the autophagy inhibitor 3-MA reversed the analgesic effect of UNC5B. In cultured SCs, UNC5B helped recruit netrin-1 to the cell membrane. UNC5B overexpression promoted autophagic flux while inhibiting apoptosis, which was further augmented with exogenous netrin-1 and reversed by netrin-1 knockdown. The enhanced phosphorylation of AMP-activated protein kinase (AMPK) and Unc51-like autophagy activating kinase 1 (ULK1) by UNC5B overexpression was also correlated with netrin-1. Our results suggest that UNC5B facilitates autophagic flux in SCs via phosphorylation of AMPK and ULK1, dependent on its ligand netrin-1, protecting myelin and partly preventing injury-induced NP.

Mammalian Sterile 20-Like Kinase 1 Mediates Neuropathic Pain Associated with Its Effects on Regulating Mitophagy in Schwann Cells.

Myelin degradation initiated by Schwann cells (SCs) after nerve injury is connected to the induction and chronicity of neuropathic pain (NP). Mitophagy, a selective clearance of damaged mitochondria via autophagy, contributes to the maintenance of normal function in SCs. Mitochondrial function and mitophagy activity are highly modulated by mammalian ste20-like kinase1 (Mst1). However, whether Mst1 can regulate mitophagy in SCs to play a role in NP remains poorly understood. In the present study, Sprague-Dawley rats were subjected to chronic constriction injury (CCI) on the sciatic nerve to induce NP. Small interfering RNA of Mst1 was applied to the injured sciatic nerve to knockdown Mst1. Behavioral tests were performed to evaluate NP, and myelin degeneration was assessed by transmission electron microscope and immunofluorescence. Autophagy and mitophagy were detected in the injured sciatic nerve and cultured SCs (RSC96 cells) by Western blot. ROS level, mitochondria membrane potential, and apoptosis were assessed in vitro via flow cytometry and Western blot. Mst1 knockdown alleviated mechanical allodynia and thermal hyperalgesia in the CCI-induced NP model and rescued myelin degeneration of the injured nerve. Meanwhile, CCI-increased levels of Parkin and p62 were reversed by Mst1 knockdown. In vitro RSC96 cells were subjected to starvation to induce mitophagy. Protein levels of mitochondrial Parkin and mitochondrial p62 significantly increased after Mst1 knockdown, while those in the cytosol diminished indicate that the translocation of Parkin and p62 from the cytosol to the mitochondria was promoted by the knockdown of Mst1. In addition, Mst1 knockdown reduced ROS level and apoptosis activity, while enhancing mitochondria membrane potential in RSC96 cells. The study showed that Mst1 knockdown alleviated CCI-induced NP, associated with enhanced Parkin recruitment to mitochondria and subsequent mitophagy degradation, thus preserving mitochondrial function and myelin integrity.

Anti-GQ1b Antibody Syndrome with Visual Impairment: A Retrospective Case Series.

BACKGROUND: Anti-GQ1b antibody syndrome referred to a clinical spectrum characterized by acute onset of ataxia, ophthalmoplegia and areflexia, while visual deterioration was rarely reported in terms of ocular disorders. This study aimed to describe the clinical characteristics of anti-GQ1b antibody syndrome with visual impairment. METHODS: The database at the First Affiliated Hospital of Sun Yat-sen University was searched from 2014 to 2020. Patients with anti-GQ1b IgG were identified and divided into two groups according to the existence of optic neuropathy. Clinical and laboratory data of these subjects between the two groups were collected and analyzed. All patients were followed up by telephone to assess the outcome. RESULTS: A total of 12 patients with seropositive anti-GQ1b antibody were included, 75% of which got antecedent infection. Of these cases, 3 showed visual deterioration accompanied by abnormal orbital magnetic resonance imaging or visual evoked potentials, and the other 9 didn't show any evidence of vision impairment. Patients in the optic neuropathy group presented prominent visual impairments as initial symptoms and were more likely to suffer from facial weakness. There were 4 patients in normal visual acuity group complaining of blurred vision due to intraocular muscle paralysis, which was distinguished by subsequent examination. The combination of glucocorticoids and intravenous immunoglobulin was applied to treat patients with optic neuropathy. CONCLUSIONS: This study provides strong evidence that anti-GQ1b antibody syndrome can exhibit visual impairment, which helps further expand the clinical spectrum of anti-GQ1b antibody syndrome. More attention should be paid to the physical and supplementary ophthalmological examination to explore the pathogenesis and treatment of anti-GQ1b antibody syndrome.

The Physiological Significance of A-Waves in Early Diabetic Neuropathy: Assessment of Motor Nerve Fibers by Neurophysiological Techniques.

Objective: This study aimed to investigate how early A-waves could occur in type II diabetes, and what it implied functionally. Methods: We performed conduction velocity distribution (CVD) test in peroneal nerves of 37 type II diabetic patients with normal nerve conduction study (NCS) and 22 age-matched controls. The electrophysiological data and clinical information were analyzed. Results: A-waves were observed in 45.9% of diabetic patients and only in 1 person in healthy controls, all detected in the tibial nerves. The diabetic patients with A-waves showed faster conduction velocity in all quartiles in the motor peroneal nerves compared to the patients without A-waves, and their CVD histograms were shifted to the right side, consisting of a significantly larger percentage of fast conducting fibers. There was no significant difference in the CVD values of the upper extremity nerves among the patients with and without A-waves and the healthy controls. Conclusion: A-waves could occur in type II diabetes as early as when NCS showed normal, and represented as a sign of neuropathy as well as a sign of rescued motor nerve function.

Local uncoordinated gene 5H2 contributes to nerve injury-induced mechanical allodynia associated to its role in autophagy.

Lesions of the peripheral nerves can lead to lifelong neuropathic pain (NP). Autophagic deficiency in the Schwann cells (SCs) is an early event in the origin of NP chronification. Uncoordinated gene 5H2 (UNC5H2), one of the repulsive netrin receptors, mediated the effect of netrin-1 on autophagic activation and cell survival in endothelial cells. However, its role on autophagy regulation in peripheral nerves during NP process remains unidentified. Chronic constriction injury (CCI) of the left sciatic nerve was induced in Sprague-Dawley rats, and UNC5H2 small interfering RNA was transfected to the ipsilateral sciatic nerve immediately after injury. Mechanical allodynia was assessed. Sciatic UNC5H2 and netrin-1 protein levels were investigated. Autophagy in the ipsilateral sciatic nerves was evaluated by detecting punctate light chain 3(LC3) and autophagosomes, as well as the levels of LC3 II, p62 and phosphorylated UNC51-like kinase (ULK1). After CCI, UNC5H2 of the sciatic nerves was upregulated, exclusively expressed in SCs. Small interfering RNA transfection resulted in significant decrease of UNC5H2 and netrin-1 protein, leading to exaggeration of mechanical allodynia through 14 days after CCI. Autophagy was activated but autophagic influx was interfered within a week after CCI, shown by the elevated levels of both LC3II and p62, which was further deteriorated with UNC5H2 knockdown. In addition, the injury-induced augmentation of phosphorylated ULK1 was significantly diminished by UNC5H2 knockdown. Altogether, the results suggest that local UNC5H2 of the peripheral nerve plays a significant role in the process of injury-induced mechanical allodynia, probably associated to its contribution to autophagic regulation.

The rostral to caudal gradient of clinical and electrophysiological features in sporadic amyotrophic lateral sclerosis with bulbar-onset.

OBJECTIVE: Amyotrophic lateral sclerosis (ALS) with bulbar-onset (BO-ALS) tends to propagate to the adjacent anatomical regions symptomatically. However, the spreading pattern of clinical and electrophysiological features is not well documented. METHODS: This retrospective study enrolled consecutive patients with sporadic BO-ALS. The clinical progression and electrophysiological data by electromyography examination were retrospectively analysed based on information from the medical records. RESULTS: The study enrolled 57 patients: 43 presented with contiguous (37 of 57) or non-contiguous (6 of 57) progression clinically; and 14 patients did not present with symptomatic propagation to other spinal segments. Lower motor neuron dysfunction was more frequently involved in the bulbar and cervical segments and less in the thoracic and lumbosacral segments. As a result, a small proportion of patients had intact thoracic paraspinal or leg muscles or both by electromyography examination. Furthermore, the patients with diagnostic latency ≤6 months showed a significantly lower incidence of neurogenic changes in the lumbosacral spinal cord compared with those with diagnostic latency > 6 months. CONCLUSION: This current study demonstrated a relative rostral-caudal descending gradient of lower motor neuron dysfunction in patients with BO-ALS. These results suggest that follow-up EMG might be necessary for a proportion of patients.