Fast Blue Staining Research Articles

Single-nucleus transcriptome profiling of prefrontal cortex induced by chronic methamphetamine treatment

BackgroundMethamphetamine (METH) addiction causes a huge burden on society. The prefrontal cortex (PFC), associated with emotion and cognitive behaviours, is also involved in addiction neurocircuitry. Although bulk RNA sequencing has...

Effect of fluoxetine treatment on neurotoxicity induced by lysolecithin in male rats.

Demyelination disorder is an unusual pathologic event, which occurs in the central nervous system (CNS). Multiple sclerosis (MS) is an inflammatory demyelinating disease that affects the CNS, and it is the leading cause of disability in young adults. Lysolecithin (LPC) is one of the best toxin-induced demyelination models. In this study, a suitable model is created, and the effect of fluoxetine treatment is examined on this model. In this case, it was assumed that daily fluoxetine treatment had increased the endogenous remyelination in the LPC model. This study was focused on investigating the influence of the fluoxetine dose of 5 or 10mg/kg per day for 1 and 4 weeks on LPC-induced neurotoxicity in the corpus callosum region. It was performed as a demyelinating model in male Wistar rats. After 3 days, fluoxetine was injected intraperitoneally (5 or 10mg/kg per day) for 1 and 4 weeks in each group. After completing the treatment course, the corpus callosum was removed to examine the gene expression and histological analysis was performed. The results of the histopathological study of hematoxylin and eosin staining of the corpus callosum showed that in 1 and 4-week treatment groups, fluoxetine has reduced the level of inflammation at the LPC injection site (5 and 10mg/kg per day). Fluoxetine treatment in the luxol fast blue (LFB) staining of the corpus callosum has been led to an increase in myelination capacity in all doses and times. The results of the genetic study showed that the fluoxetine has significantly reduced the expression level of tumor necrosis factor-α, nuclear factor κβ, and induced nitric oxide synthase in comparison with the untreated LPC group. Also, the fluoxetine treatment has enhanced the expression level of the forkhead box P3 (FOXP3) gene in comparison with the untreated group. Fluoxetine has increased the expression level of myelination and neurotrophic genes such as myelin basic protein (MBP), oligodendrocyte transcription factor 2 (OLIG2), and brain-derived neurotrophic factor (BDNF). The outcomes demonstrated that fluoxetine reduces inflammation and strengthens the endogenous myelination in the LPC-induced demyelination model; however, supplementary studies are required for specifying the details of its mechanisms.

LRRC4 functions as a neuron-protective role in experimental autoimmune encephalomyelitis

BackgroundLeucine rich repeat containing 4 (LRRC4), also known as netrin-G ligand-2 (NGL-2), belongs to the superfamily of LRR proteins and serves as a receptor for netrin-G2. LRRC4 regulates the formation of excitatory synapses and promotes axon differentiation. Mutations in LRRC4 occur in Autism Spectrum Disorder (ASD) and intellectual disability. Multiple sclerosis (MS) is a chronic neuroinflammatory disease with spinal cords demyelination and neurodegeneration. Here, we sought to investigate whether LRRC4 is involved in spinal cords neuron-associated diseases.MethodsLRRC4 was detected in the CNS of experimental autoimmune encephalomyelitis (EAE) mice by the use of real-time PCR and western blotting. LRRC4−/− mice were created and immunized with myelin oligodendrocyte glycoprotein peptide (MOG)35–55. Pathological changes in spinal cords of LRRC4−/− and WT mice 15 days after immunization were examined by using hematoxylin and eosin (H&E), Luxol Fast Blue (LFB) staining and immunohistochemistry. The number of Th1/Th2/Th17/Treg cells in spleens and blood were measured with flow cytometry. Differential gene expression in the spinal cords from WT and LRRC4−/− mice was analyzed by using RNA sequencing (RNA-seq). Adeno-associated virus (AAV) vectors were used to overexpress LRRC4 (AAV-LRRC4) and were injected into EAE mice to assess the therapeutic effect of AAV-LRRC4 ectopic expression on EAE.ResultsWe report that LRRC4 is mainly expressed in neuron of spinal cords, and is decreased in the spinal cords of the EAE mice. Knockout of LRRC4 have a disease progression quickened and exacerbated with more severe myelin degeneration and infiltration of leukocytes into the spinal cords. We also first found that Rab7b is high expressed in EAE mice, and the deficiency of LRRC4 induces the elevated NF-κB p65 by up-regulating Rab7b, and up-regulation of IL-6, IFN-γ and down-regulation of TNF-α, results in more severe Th1 immune response in LRRC4−/− mice. Ectopic expression of LRRC4 alleviates the clinical symptoms of EAE mice and protects the neurons from immune damages.ConclusionsWe identified a neuroprotective role of LRRC4 in the progression of EAE, which may be used as a potential target for auxiliary support therapeutic treatment of MS.

Young plasma attenuates cognitive impairment and the cortical hemorrhage area in cerebral amyloid angiopathy model mice.

BackgroundCerebral amyloid angiopathy (CAA) is characterized by the deposition of β-amyloid (Aβ) in leptomeningeal vessels and penetrating arterioles. Intracerebral hemorrhage (ICH) is one of the most destructive complications in CAA. Young plasma has been shown to improve cognitive, learning, and memory functions in Alzheimer’s disease (AD) model mice and is a new potential therapy. However, it is not clear whether young plasma can reduce cerebral hemorrhage and improve the prognosis of neurological function in APP/PS1 (which express APP695swe and PS1-dE9 mutations) mice with CAA disease.MethodsThe Y-maze, new object recognition (NOR), forced swimming, open field, sucrose consumption, and corner tests were used to evaluate the learning and memory, cognitive ability, and emotional changes in CAA model mice. The effect of young plasma on neurogenesis was analyzed by immunofluorescence. The level of Aβ in the cerebral cortex and hippocampus of mice was measured by enzyme-linked immunosorbent assay (ELISA). Finally, the area of cortical hemorrhage in mice was analyzed by fast blue-staining.ResultsWe proved that young plasma improved cognition, learning and memory impairment, and anxiety in CAA model mice, prevented neuronal apoptosis, and enhanced neurogenesis in APP/PS1 mice. However, young plasma did not reduce the level of Aβ in the cortex and hippocampus of APP/PS1 mice. We also found that young plasma reduced the area of cerebral hemorrhage in APP/PS1 mice.ConclusionsOur results show that young plasma can improve learning and memory, cognitive impairment, and anxiety in CAA model mice and can reduce the area of cortical hemorrhage.

Host tissue response to floating microelectrode arrays chronically implanted in the feline spinal nerve

Objective. Neural interfacing technologies could significantly improve quality of life for people living with the loss of a limb. Both motor commands and sensory feedback must be considered; these complementary systems are segregated from one another in the spinal nerve. Approach. The dorsal root ganglion-ventral root (DRG-VR) complex was targeted chronically with floating microelectrode arrays designed to record from motor neuron axons in the VR or stimulate sensory neurons in the DRG. Hematoxylin and eosin and Nissl/Luxol fast blue staining were performed. Characterization of the tissue response in regions of interest and pixel-based image analyses were used to quantify MAC387 (monocytes/macrophages), NF200 (axons), S100 (Schwann cells), vimentin (fibroblasts, endothelial cells, astrocytes), and GLUT1 (glucose transport proteins) reactivity. Implanted roots were compared to non-implanted roots and differences between the VR and DRG examined. Main results. The tissue response associated with chronic array implantation in this peripheral location is similar to that observed in central nervous system locations. Markers of inflammation were increased in implanted roots relative to control roots with MAC387 positive cells distributed throughout the region corresponding to the device footprint. Significant decreases in neuronal density and myelination were observed in both the VR, which contains only neuronal axons, and the DRG, which contains both neuronal axons and cell bodies. Notably, decreases in NF200 in the VR were observed only at implant times less than ten weeks. Observations related to the blood-nerve barrier and tissue integrity suggest that tissue remodeling occurs, particularly in the VR. Significance. This study was designed to assess the viability of the DRG-VR complex as a site for neural interfacing applications and suggests that continued efforts to mitigate the tissue response will be critical to achieve the overall goal of a long-term, reliable neural interface.

NAD+ attenuates experimental autoimmune encephalomyelitis through induction of CD11b+ gr-1+ myeloid-derived suppressor cells.

Objective: To investigate the effects of nicotinamide adenine dinucleotide (NAD+) on the pathogenesis of the animal model for multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE).Methods: EAE model was induced by myelin oligodendrocyte protein (MOG 35-55). Clinical scores of EAE were measured in mice with or without NAD+ treatment. Hematoxylin and Eosin (HE) and Luxol Fast Blue (LFB) staining were performed to assess inflammation and demyelination, respectively. Expressions of target proteins were measured by Western blot. The numbers of myeloid-derived suppressor cells (MDSCs) were measured by immunofluorescent staining and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to measure the expressions of inflammatory cytokine in serum.Results: NAD+ treatment could decrease inflammatory cells and demyelination foci, attenuate the clinical scores of EAE and slightly delay disease onset. Western blot showed that NAD+ treatment up-regulated the expression of phosphorylated-STAT6 (p-STAT6) and SIRT1. Besides, NAD+ treatment up-regulated the expression of p-IκB and down-regulated the expression of p-NF-κB. In addition, NAD+ treatment could increase the numbers of CD11b+ gr-1+ MDSCs and the expression of Arginase-1. Moreover, NAD+ treatment up-regulated the expressions of IL-13 and down-regulated the expression of IFN-γ and IL-17.Conclusions: The present study demonstrated that NAD+ treatment may induce the CD11b+ gr-1+ MDSCs to attenuate EAE via activating the phosphorylation of STAT6 expression. Therefore, NAD+ should be considered as a potential novel therapeutic strategy for MS.

Ellagic acid improves muscle dysfunction in cuprizone-induced demyelinated mice via mitochondrial Sirt3 regulation

Sirt3 enzyme and mitochondrial abnormality can be related to excess fatigue or muscular dysfunction in multiple sclerosis (MS).Ellagic acid (EA) has a mitochondrial protector, iron chelator, antioxidant, and axon regenerator in neurons.In this study the effect of EAon muscle dysfunction, its mitochondria, and Sirt3 enzyme incuprizone-induced model of MSwas examined.Demyelination was induced by a diet containing 0.2% w/w cuprizone (Cup) for 42 days and EA administered daily (5, 50, and 100 mg/kg P.O) either with or without cuprizone in mice. Behavioral tests were assessed, and muscle tissue markers ofoxidative stress, mitochondrial parameters, mitochondrial respiratory chain activity, the Sirt3 protein level, and Sirt3 expression were also determined.Luxol fast blue staining and the behavioral tests were performed toassess the implemented model. In Cup group an increased oxidative stress in their muscle tissues was observed. Also, muscle mitochondria exhibited mitochondria dysfunction, lowered mitochondrial respiratory chain activity, Sirt3 protein level, and Sirt3 expression.EA prevented most of these anomalous alterations.Sub-chronicEA co-treatment dose-dependently ameliorated behavioral and muscular impairment in mice that received Cup.EA can effectively protect muscle tissue against cuprizone-induced demeylination via the mitochondrial protection, oxidative stress prevention and Sirt3 overexpression.

Abstract # 2051 Chemobrain in rats – Morphological, oxidative and behavioral effects of doxorubicin administration

Doxorubicin (Dox), a topoisomerase interactive agent, is commonly used to treat several types of cancer. This drug is known to cause cognitive impairments in individuals submitted to long-term chemotherapy (deficits also called as chemobrain). The present study investigated whether morphological, oxidative and behavioral impairments could be induced by Dox. Male Wistar rats were injected with Dox (2.5 mg/kg, once every week for 28 days, intraperitoneal route-IP) or 0.9% saline solution (same volume, IP). During the experimental period, behavioral studies were performed, including the open field test, the elevated plus maze, tests for sociability and preference for social novelty and the novel object recognition task. Brains were analyzed by immunohistochemistry (GFAP or glial fibrillary acidic protein expression in astrocytes) and hematoxylin-eosin and luxol fast blue (for myelin) staining techniques, as well as for the assessment of oxidative parameters (thiobarbituric acid reactive substances-TBARS, catalase, superoxide dismutase-SOD, glutathione peroxidase-GPX, glutathione reductase-GR, nitric oxide-NO). Dox-injected rats presented increased GFAP expression in all analyzed areas (frontal cortex, striatum, hippocampus, hypothalamus, granular and molecular layers of the cerebellum). Dox administration also increased the levels of the oxidative indicators TBARS, NO and GR and caused memory impairments as seen in the novel object recognition test. No signs of demyelination and/or neuronal loss were found. Results suggest that astrogliosis and oxidative stress induced by Dox may be linked to chemotherapy-induced memory deficits.

Expression and significance of estrogen receptor in bladder of experimental autoimmune encephalomyelitis mouse model

Objective To study the changes of estrogen receptor (ER) expression in neurogenic bladder caused by multiple sclerosis (MS) and its significance by establishing an experimental autoimmune encephalomyelitis (EAE) model in mice. Methods The EAE models were induced by an antigen emulsion mixture that was well mixed with mog35-55 and complete Freund’s adjuvant. Twenty female C57Bl/6 mice were randomly divided into experimental group and control group. The clinical symptoms of mice were observed and recorded after immunization. Twenty-third days after immunization, micturition frequency and mean void weight were observed within 4 h. The spinal cord as well as the bladder was collected and the bladder was weighed. The pathological changes were observed by hematoxylin-eosin (HE) staining and Luxol fast blue (LFB) staining. The expression of ER in bladder of EAE mice was detected by Western blotting. Results The clinical score of neurological function in EAE mice increased, and typical MS symptoms appeared. The weight of the control group and experimental group was (23.0±1.1) g and (16.5±1.5) g respectively. As compared with the control group, the weight of the mice in the model group was obviously reduced. The average number of 4H urination in model group (8.7±1.4) was significantly greter than in the control group (4.2±1.1, P=0.001). The urine volume in the model group [(0.22±0.11) mg] was significantly less than that in the control group [(0.34±0.07) mg, P=0.008]. In the model group, the bladder weight ratio [(0.26±0.09)‰] was significantly higher than that in the model group [(0.11±0.03)‰, P=0.001]. HE-LFB staining showed obvious inflammatory cell infiltration and demyelination in the spinal cord of EAE model mice, and Western blotting showed that the relative expression of ER in the bladder of model group (0.38±0.08) was significantly lower than that in the control group (0.71±0.19, P=0.001). Conclusion Mog35-55 can successfully induce the EAE mice model. The EAE mice model can well simulate the MS neurogenic bladder symptoms. ER and estrogen may be the important factors in bladder dysfunction caused by multiple sclerosis. Key words: Estrogen receptor; Experimental autoimmune encephalomyelitis; Neurogenic bladder

Histopathological effects of intramuscular metamizole sodium on rat sciatic nerve.

We investigated the histopathological effects of metamizole sodium (MS) on the sciatic nerve. This study was performed using 48 adult male Wistar albino rats. Ten groups were constituted with 6 rats in each group. MS injection into the sciatic nerve (group 1), MS injection into the muscle [group 3 (50 mg/kg, 0.4 ml) and group 5 (50 mg/kg, 0.8 ml)], MS injection into the muscle cavity in the vicinity of the sciatic nerve [group 2 (50 mg/kg, 0.4 ml) and group 4 (50 mg/kg, 0.8 ml)], normal saline injection into the muscle in the vicinity of the sciatic nerve [group 6A (0.4 ml) and 6B (0.8 ml)], subjected to injury by drilling the entire layer of nerve without injecting any drug, normal saline injection in the sciatic nerve, and control group. Nerve and muscle samples were taken 7 days after administrations. Tissue sections were stained using a hematoxylin and eosin-Luxol® fast blue stain, assessed by a histologist. The levels of axonal degeneration of the rats in groups 1, 2, 3, 4, 5, 6A, and 8 were found to be significantly higher compared to the levels of the rats in the control group (P<0.05). Myelin degeneration of the rats in all groups was found to be significantly higher compared to myelin degeneration of the rats in the control group (P<0.05). It was observed that MS could lead to injury in the sciatic nerve with a toxic effect due to diffusion.

Alpha-tocopherol ameliorates experimental autoimmune encephalomyelitis through the regulation of Th1 cells.

Multiple sclerosis (MS) is a serious neurological autoimmune disease, it commonly affects young adults. Vitamin E (Vit E) is an important component of human diet with antioxidant activity, which protects the body's biological systems. In order to assess the effect of Vit E treatment on this autoimmune disease, we established experimental autoimmune encephalomyelitis (EAE), the animal model of MS, and treated EAE with α-tocopherol (AT) which is the main content of Vit E. Twenty C57BL/6 adult female mice were used and divided into two groups randomly. EAE was induced with myelin oligodendrocyte glycoprotein (MOG), and one group was treated with AT, at a dose of 100 mg/kg on the 3(th) day post-immunization with MOG, the other group was treated with 1% alcohol. Mice were euthanized on day 14, post-immunization, spleens were removed for assessing splenocytes proliferation and cytokine profile, and spinal cords were dissected to assess the infiltration of inflammatory cells in spinal cord. AT was able to attenuate the severity of EAE and delay the disease progression. H&E staining and fast blue staining indicated that AT reduced the inflammation and the demyelination reaction in the spinal cord. Treatment with AT significantly decreased the proliferation of splenocytes. AT also inhibited the production of IFN-γ (Th1 cytokine), though the other cytokines were only affected slightly. According to the results, AT ameliorated EAE, through suppressing the proliferation of T cells and the Th1 response. AT may be used as a potential treatment for MS.

Abstract WMP38: ABCA1/HDL Pathway Mediates GW3965 Induced Neurorestoration and White Matter Remodeling After Stroke

ATP-binding cassette transporter A1 (ABCA1), a major cholesterol transporter in the central nervous system, plays an important role in the formation of brain HDL-cholesterol. ABCA1 is up-regulated by the transcription factors, liver X receptors (LXRs). Our previous study using brain-specific ABCA1 deficient (floxed, nestin-Cre positive, ABCA1-B/-B) and ABCA1 floxed-control (ABCA1fl/fl) mice showed that ABCA1 has anti-inflammatory effects and reduces white matter (WM) damage in ischemic stroke models. Here, we further test whether GW3965, a synthetic LXR agonist treatment of stroke increases ABCA1 and HDL, and thereby improves WM-remodeling and functional outcome. Adult male ABCA1fl/fl and ABCA1-B/-B mice were subjected to permanent extraluminal distal middle cerebral artery occlusion (dMCAo) by transcranial ligation method. Animals were gavaged with saline or GW3965 (10 mg/kg) starting 24h after dMCAo and daily till sacrificed 14 days after dMCAo. There were no differences in the lesion volume and blood levels of total cholesterol and triglyceride before and after dMCAo between ABCA1-B/-B and ABCA1fl/fl mice. However, GW3965 treatment significantly increased blood HDL levels in ABCA1fl/fl mice, but not in ABCA1-B/-B mice 14 days after dMCAo. GW3965 treatment of ABCA1fl/fl stroke mice, but not in ABCA1-B/-B stroke mice, also increased: 1) WM-density identified by Luxol Fast Blue (myelin marker), Bielshowsky silver (axon marker), and SMI31 (phosphorylated neurofilament marker) staining; 2) the number of platelet-derived growth factor receptor alpha positive oligodendrocyte progenitor cells; 3) ABCA1, synaptophysin and myelin basic protein expression in the ischemic brain; 4) functional outcome 7 and 14 days after dMCAo, compared with non-treatment ABCA1fl/fl stroke mice (p&lt;0.05, n=9/group). GW3965 and HDL treatment significantly increase axonal/neurite outgrowth in cultured primary cortical neurons derived from ABCA1fl/fl embryos, but not in neurons derived from ABCA1-B/-B embryos. Treatment of stroke with GW3965 significantly increased blood level of HDL, increased ABCA1 expression and WM-remodeling in the ischemic brain. Increasing ABCA1 may contribute to GW3965 induced neurorestoration and WM remodeling after stroke.

MHC-mismatched mixed chimerism augments thymic regulatory T cell production and prevents relapse of EAE (THER7P.960)

Abstract Multiple sclerosis (MS) is an autoimmune inflammatory disease in central nervous system with demyelination, axon damage, and paralysis. There is no yet curative therapy. Induction of MHC-mismatched mixed chimerism is proposed to be a potential cure for autoimmune diseases, but this has not yet been tested in animal models of MS, experimental autoimmune encephalomyelitis (EAE). Here, we report that MHC-mismatched mixed chimerism with C57BL/6 (H-2b) donor in SJL/J (H-2s) EAE recipients eliminates symptoms and prevents relapse. This cure is demonstrated by not only disappearance of clinical signs but also reversal of autoimmunity and elimination of infiltration of T, B and macrophage cells in the spinal cord, as judged by HE, immunofluorescent staining and flow cytometry analysis. We also see regeneration of myelin as judged by fast-blue staining and prevention of axon damage as judged by electronic microcopy. The reversal of autoimmunity is associated with a marked reduction of autoreactivity of CD4+ T cells and significant increase of Foxp3+ regulatory T percentage among host-type CD4+ T cells in the spleen and lymph nodes. We have also found a marked increase of Foxp3+ regulatory T percentage among host-type CD4+ and CD4+CD8+ thymocytes in the thymus. Thymectomy leads to loss of prevention of EAE relapse by the mixed chimerism. These results indicate that induction of MHC-mismatched mixed chimerism can augment thymic production of Foxp3+ regulatory T cells and cure EAE.

Role of new K(+) channel blocker 4-AP-3-MeOH in acute spinal cord compression injury in rats

To evaluate the effects of 4-aminopyridine-3-methyl hydroxide (4-AP-3-MeOH) in rat's acute spinal cord injury. A total of 12 adult male SD rats (250-300 g) were randomly divided into treatment (n = 6) and control (n = 6) groups. After compressing segment T11 of spinal cord for 30 min, the injured segment received 1 ml 4-AP-3-MeOH (100 µmol/ml) by topically application in treatment group while the control group received 1 ml saline.Somatosensory evoked potential (SSEP) was detected in both groups at pre-injury, 30 min post-injury and post-dosing. Then Luxol fast blue (LFB) staining of target spinal segment was performed. In treatment group, the values of SSEP at pre-injury, 30 min post-injury and post-dosing were 1.26 ± 0.35, 0.03 ± 0.05 and 0.45 ± 0.19 µv respectively. Comparing SSEP of 30 min post-injury with post-dosing, the difference was statistically significant (P < 0.01).While in control group, the values of SSEP at pre-injury, 30 min post-injury and post-dosing were 1.05 ± 0.39, 0.01 ± 0.02 and 0.02 ± 0.02 µv respectively. Comparing SSEP of 30 min post-injury with post-dosing, there was no statistical difference (P > 0.05). After 30 min injury, there were swelling and bleeding of spinal cord.LFB staining showed that both gray and white matter had swelling and bleeding and central canal was destroyed with varying degrees of demyelination. After 30 min of acute spinal cord injury, there are bleeding of gray and white matter with varying degrees of demyelination. Topical usage of K(+) blocker 4-AP-3-MeOH can effectively improve the conduction of SSEP after acute spinal cord injury in rats.

Toll like receptors signaling pathways directly increase the expression of functional IL-17RA in neu-roglial cells

Objective To investigate whether toll like receptor （ TLR） signaling pathways can in-crease the expression of IL-17 R in neuralglial cells , and if they can whether the increased IL-17 R is func-tional.Methods Experimental autoimmune encephalomyelitis （EAE） was induced in B6 mice by immuni-zation with an emulsion of myelin oligodendrocyte glycoprotein 35-55 （ MOG35-55 ） in complete Freund’s adju-vant （CFA）.The expression of Il17ra and Il17rc in the brains and spinal cords of mice with EAE were de-tected by real-time PCR.Luxol fast blue （ LFB） staining was performed to the spinal cord sections to detect tissue demyelination.Immunohistological staining against IL-17RA and CD3 were undertook to visualize IL-17RA＋and CD3 ＋ cells.Same approaches were also applied to immunized Rag1 -/- mice to figure out whether T cells infiltration is necessary for increasing IL-17RA expression in the central nervous system （ CNS） .Then B6 mice were immunized with incomplete Freund′s adjuvant （ IFA） plus different TLRs ago-nists to measure the expression of Il17ra in the brains and spinal cords by qPCR .The purified astrocytes , microglia and oligodendrocytes isolated from neonatal mice brains were cultured in vitro for two weeks , and then treated with different TLRs agonists .The expression of Il17ra at mRNA and protein levels in the cells were determined by qPCR and Western blot respectively .The astrocytes were treated with IL-17A and LPS individually or in the combination to detect the level of CCL 2, CXCL8 and IP-10 in the supernatant by ELISA.Results B6 mice with induced EAE showed significantly increased Il17ra expressions in the brain＆amp;nbsp;and spinal cord , which was also detected in immunized Rag1 -/-mice.Although no spinal cord demyeliza-tion and CD3 cells infiltration were detected in Rag1 -/-mice, significantly increased number of IL-17RA positive cells could still be visualized .In vivo TLRs agonist participated immunization and in vitro treatment of purified neuroglial cells demonstrated that TLRs agonists could directly evoke IL -17RA expression in the CNS or cultured astrocytes , microglia and oligodendrocytes with high efficiency .Both IL-17 A and LPS could stimulate astrocytes to secrete CCL2, CXCL8 and IP-10, however, a combined use of IL-17A and LPS fur-ther augmented the production of these chemokines to a large extend .Conclusion Taken together , we con-cluded that TLRs agonists could directly stimulate neuroglial cells to express IL -17RA which functionally re-spond to IL-17A by secreting chemokines . Key words: Toll-like receptor; Neuroglial cells; IL-17 receptor; Expression

PD-L1 is increased in the spinal cord and infiltrating lymphocytes in experimental allergic encephalomyelitis.

Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Th1 cells play an important role in the pathogenesis of experimental allergic encephalomyelitis. This study determined the potential effect of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein, complete Freund's adjuvant and Bordetella pertussis toxin into C57BL/6J mice. Experimental allergic encephalomyelitis mice developed disease and showed inflammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections, demyelination by Luxol fast-blue staining and clinical manifestations. The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry, flow cytometry and western blot analysis. The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice. Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.

Human umbilical cord Wharton's jelly-derived oligodendrocyte precursor-like cells for axon and myelin sheath regeneration.

Human umbilical mesenchymal stem cells from Wharton's jelly of the umbilical cord were induced to differentiate into oligodendrocyte precursor-like cells in vitro. Oligodendrocyte precursor cells were transplanted into contused rat spinal cords. Immunofluorescence double staining indicated that transplanted cells survived in injured spinal cord, and differentiated into mature and immature oligodendrocyte precursor cells. Biotinylated dextran amine tracing results showed that cell transplantation promoted a higher density of the corticospinal tract in the central and caudal parts of the injured spinal cord. Luxol fast blue and toluidine blue staining showed that the volume of residual myelin was significantly increased at 1 and 2 mm rostral and caudal to the lesion epicenter after cell transplantation. Furthermore, immunofluorescence staining verified that the newly regenerated myelin sheath was derived from the central nervous system. Basso, Beattie and Bresnahan testing showed an evident behavioral recovery. These results suggest that human umbilical mesenchymal stem cell-derived oligodendrocyte precursor cells promote the regeneration of spinal axons and myelin sheaths.

Characterization of the tissue response to functional multielectrode arrays in the feline spinal nerve

Peripheral nervous system sites are target implantation regions for neural prosthetics that restore function or provide therapy. Despite advances in electromechanical prostheses, current peripheral‐interface technology cannot convey the users’ intent to the prosthesis. Anatomical information and an understanding of the tissue response are needed. In this study, functional microelectrode arrays (FMAs) were implanted into feline dorsal (sensory) or ventral (motor) roots. At predetermined time points or when &lt;10% of electrodes had recordable signals, animals were perfused and the implanted tissue formalin‐fixed and paraffin‐embedded. Hematoxylin and eosin and Nissl/Luxol fast blue staining along with antibody‐based stains [NF200 (axons), S100 (Schwann cells), vimentin (fibroblasts, endothelial cells, astrocytes), NeuN (neuronal nuclei), caspase‐3 (apoptotic cell death), MAC387 (monocytes/macrophages) and occludin (tight junctions/blood‐nerve barrier integrity)] were used. Axonal density, scar formation, neural degeneration (ventral root and spinal cord ventral horn) and inflammation were quantified and compared in implanted and non‐implanted roots. Histological data was correlated with functional metrics to provide insight toward the overall goal of a long‐term, reliable interface with prosthetic limbs. Funding provided by the DARPA RPI grant.

Abstract TMP36: Persistent Stroke-induced Sensory, Motor And Cognitive Deficits Involve Long-term Myelin Loss In Major Fiber Tracts

Objectives: Persistent neurobehavioral deficits and brain changes need validation for future brain restoration research. Here neurological and brain changes were determined post-stroke for this purpose. Methods: Two hr middle cerebral artery occlusion (tMCAO) or sham (Sham) surgery was performed in male SD rats. Measurements made over 10 weeks included: (1) sensory, motor, beam balance, hindlimb and forepaw behaviors, and (2) complex active place avoidance learning (APA) and simple passive avoidance retention (PA). Electroretinogram (ERG) was measured at term. Measurements of hemispheric loss (infarction), hippocampus CA1 neuronal loss and myelin (Luxol Fast Blue) staining in several fiber tracts were made. Results: Compared to Sham, tMCAO produced significant but acute deficits in forelimb foot fault for only 1 day (max 2144 ± 556 % change, p&lt;0.01) and forelimb cylinder placement for only 2 weeks (max 822 ± 215 % change, p&lt;0.01). Persistent deficits for the 10 week period were exhibited for motor, sensory and beam balance performance and for hindlimb placement (all &gt; 20-fold, p&lt;0.001). Motor behavior and beam balance deficits, although persistent, did decrease 30-40 % over time. tMCAO produced significant, cognitive deficits in APA learning but did not affect PA retention. The impaired APA learning (max 604 ± 83 % change, p&lt;0.001) was prolonged, and in fact exhibited progressive worsening (&gt; 2-fold) over time, thus being most sensitive to brain injury. ERG indicated no difference in visual function. Percent hemispheric loss measured at term was cross-validated by 2 different staining methods (33 ± 4 % for H&amp;E and 34 ± 4 % for TTC). No hippocampus neuronal loss occurred. Fiber tract myelin loss was significant (&gt; 2-fold, P&lt;0.01) in the external capsule and striatum but not in the corpus callosum or anterior commissure. Conclusions: Persistent deficits were identified for neurobehavioral end points for use in future restoration research. Fiber myelin loss in the external capsule and striatum contribute to long term deficits, apparently especially important for cognitive control necessary in complex learning. Investigation of myelin protection/regrowth mediated by oligodendrocytes, and perhaps other cells, will be essential to future stroke research.

Striatal neuronal loss correlates with clinical motor impairment in Huntington's disease

Huntington's disease (HD) is characterized clinically by chorea, motor impairment, psychiatric manifestations, and dementia. Atrophy of the striatum is the neuropathological hallmark of HD, and previous studies have suggested that striatal atrophy correlates more closely with motor impairment than with chorea. Motor impairment, as measured by motor impairment score, correlates with functional disability in HD patients, but chorea does not. In this study, we investigated the relation between neuronal loss and these motor features. We conducted neuropathological and stereologic assessments of neurons in putamen and subthalamic nuclei in HD patients and age-matched controls. In putamen, we estimated the total number and volume of medium spiny neurons labeled with dopamine- and cAMP-regulated phosphoprotein 32 kDa (DARPP-32). In subthalamic nuclei, we estimated the total number of neurons on hematoxylin & eosin/luxol fast blue stains. In putamen of HD, immunohistochemistry showed DARPP-32 neuronal atrophy with extensive disruption of neurites and neuropil; stereologic studies found significant decreases in both the number and size of DARPP-32 neurons; we also detected a significant reduction of overall putamen volume in HD patients, compared to controls. In subthalamic nuclei, there was a mild, but significant, neuronal loss in the HD group. The loss of neurons in putamen and subthalamic nuclei as well as putaminal atrophy were significantly correlated with severity of motor impairment, but not with chorea. Our findings suggest that neuronal loss and atrophy in striatum and neuronal loss in subthalamic nuclei contribute specifically to the motor impairment of HD, but not to chorea.