ABSTRACT
α-Synuclein accumulates in Lewy bodies, and this accumulation is a pathological hallmark of Parkinson's disease (PD). Previous studies have indicated a causal role of α-synuclein in the pathogenesis of PD. However, the molecular and cellular mechanisms of α-synuclein toxicity remain elusive. Here, we describe a novel phosphorylation site of α-synuclein at T64 and the detailed characteristics of this post-translational modification. T64 phosphorylation was enhanced in both PD models and human PD brains. T64D phosphomimetic mutation led to distinct oligomer formation, and the structure of the oligomer was similar to that of α-synuclein oligomer with A53T mutation. Such phosphomimetic mutation induced mitochondrial dysfunction, lysosomal disorder, and cell death in cells and neurodegeneration in vivo, indicating a pathogenic role of α-synuclein phosphorylation at T64 in PD.
Subject(s)
Parkinson Disease , Humans , Parkinson Disease/metabolism , alpha-Synuclein/genetics , alpha-Synuclein/metabolism , Phosphorylation , Lewy Bodies/metabolism , Brain/metabolismABSTRACT
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by TDP-43 inclusions in the cortical and spinal motor neurons. It remains unknown whether and how pathogenic TDP-43 spreads across neural connections to progress degenerative processes in the cortico-spinal motor circuitry. Here we established novel mouse ALS models that initially induced mutant TDP-43 inclusions in specific neuronal or cell types in the motor circuits, and investigated whether TDP-43 and relevant pathological processes spread across neuronal or cellular connections. We first developed ALS models that primarily induced TDP-43 inclusions in the corticospinal neurons, spinal motor neurons, or forelimb skeletal muscle, by using adeno-associated virus (AAV) expressing mutant TDP-43. We found that TDP-43 induced in the corticospinal neurons was transported along the axons anterogradely and transferred to the oligodendrocytes along the corticospinal tract (CST), coinciding with mild axon degeneration. In contrast, TDP-43 introduced in the spinal motor neurons did not spread retrogradely to the cortical or spinal neurons; however, it induced an extreme loss of spinal motor neurons and subsequent degeneration of neighboring spinal neurons, suggesting a degenerative propagation in a retrograde manner in the spinal cord. The intraspinal degeneration further led to severe muscle atrophy. Finally, TDP-43 induced in the skeletal muscle did not propagate pathological events to spinal neurons retrogradely. Our data revealed that mutant TDP-43 spread across neuro-glial connections anterogradely in the corticospinal pathway, whereas it exhibited different retrograde degenerative properties in the spinal circuits. This suggests that pathogenic TDP-43 may induce distinct antero- and retrograde mechanisms of degeneration in the motor system in ALS.
Subject(s)
Amyotrophic Lateral Sclerosis , Retrograde Degeneration , Animals , Mice , Amyotrophic Lateral Sclerosis/pathology , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Motor Neurons/metabolism , Retrograde Degeneration/metabolism , Retrograde Degeneration/pathology , Spinal Cord/pathologyABSTRACT
INTRODUCTION: Vitamin D plays an important role in the immune system, and postnatal vitamin D insufficiency is one of the risk factors for the development of allergic disease. However, the effects of women's vitamin D intake during pregnancy on the prevalence of allergic disease in their children remain controversial. METHODS: From the Japan Environment and Children's Study, an ongoing nationwide birth cohort study, we obtained information on maternal dietary vitamin D intake determined using a food frequency questionnaire and parent-reported allergic disease symptoms based on the ISAAC questionnaire in children at 3 years of age. RESULTS: From the full dataset of 103,060 pregnancies, we analyzed complete data for 73,309 mother-child pairs. The prevalence of current wheeze, current rhinitis, current rhino-conjunctivitis, current eczema, ever asthma, ever pollinosis, and ever atopic dermatitis in the children was 17.2%, 29.7%, 3.8%, 15.2%, 9.6%, 3.7%, and 11.0%, respectively. The ORs for current rhinitis were significantly lower in the 3rd, 4th, and 5th quintiles than in the 1st quintile after adjustment for various covariates and showed a linear association. The ORs for ever pollinosis were significantly lower in the 2nd, 3rd, and 4th quintiles than in the 1st quintile, showing a U-shaped curve. There was no clear association between mothers' dietary vitamin D intake and symptoms of asthma or atopic dermatitis in their 3-year-old children. CONCLUSION: Maternal dietary vitamin D intake during pregnancy is associated with the ORs for nasal allergies in children at the age of 3 years. Further studies are warranted to evaluate the appropriate intake dose of vitamin D for pregnant women to prevent the development of nasal allergies in their children.
Subject(s)
Asthma , Dermatitis, Atopic , Rhinitis, Allergic, Seasonal , Rhinitis, Allergic , Rhinitis , Humans , Female , Pregnancy , Child, Preschool , Dermatitis, Atopic/epidemiology , Cohort Studies , Japan/epidemiology , Asthma/epidemiology , Rhinitis, Allergic/epidemiology , Vitamin DABSTRACT
BACKGROUND: Atopic march is defined as the progression from atopic dermatitis (AD) during early life to other allergic diseases in later childhood. In a nationwide birth cohort study, the Japan Environment and Children's Study, we investigated the association of bathing habits, which are known to affect skin conditions, for infants with their later development of allergic diseases. METHODS: Pregnant women who lived in 15 designated regional centers throughout Japan were recruited. We obtained information on bathing habits for their 18-month-old infants and the prevalence of allergic diseases when they were aged 3 years. RESULTS: Data for 74,349 children were analyzed. Most 18-month-old infants were bathed or showered almost every day. When they were divided into four groups according to the frequency of soap use during bathing (every time, most of the time, sometimes, and seldom), the risk of AD later at age 3 was shown to increase in association with a decreasing frequency of soap use [most of the time: adjusted odds ratio (aOR) 1.18, 95% confidence interval (CI) 1.05-1.34; sometimes: aOR 1.72, 95% CI 1.46-2.03; seldom: aOR 1.99, 95% CI 1.58-2.50], compared with soap use every time during bathing at 18 months of age. Similar results were obtained for food allergy but not for bronchial asthma. CONCLUSIONS: Frequent soap use when bathing 18-month-old infants was associated with a decreased risk of them developing allergic diseases at age 3. Further well-designed clinical studies are warranted to determine an effective bathing regimen for preventing the development of allergic diseases.
Subject(s)
Dermatitis, Atopic , Food Hypersensitivity , Pregnancy , Infant , Child , Humans , Female , Child, Preschool , Soaps , Cohort Studies , Japan/epidemiology , Prevalence , Dermatitis, Atopic/epidemiology , Dermatitis, Atopic/prevention & control , Food Hypersensitivity/epidemiologyABSTRACT
Tau is a microtubule (MT)-associated protein that is localized to the axon. In Alzheimer's disease, the distribution of tau undergoes a remarkable alteration, leading to the formation of tau inclusions in the somatodendritic compartment. To investigate how this mislocalization occurs, we recently developed immunohistochemical tools that can separately detect endogenous mouse and exogenous human tau with high sensitivity, which allows us to visualize not only the pathological but also the pre-aggregated tau in mouse brain tissues of both sexes. Using these antibodies, we found that in tau-transgenic mouse brains, exogenous human tau was abundant in dendrites and somata even in the presymptomatic period, whereas the axonal localization of endogenous mouse tau was unaffected. In stark contrast, exogenous tau was properly localized to the axon in human tau knock-in mice. We tracked this difference to the temporal expression patterns of tau. Endogenous mouse tau and exogenous human tau in human tau knock-in mice exhibited high expression levels during the neonatal period and strong suppression into the adulthood. However, human tau in transgenic mice was expressed continuously and at high levels in adult animals. These results indicated the uncontrolled expression of exogenous tau beyond the developmental period as a cause of mislocalization in the transgenic mice. Superresolution microscopic and biochemical analyses also indicated that the interaction between MTs and exogenous tau was impaired only in the tau-transgenic mice, but not in knock-in mice. Thus, the ectopic expression of tau may be critical for its somatodendritic mislocalization, a key step of the tauopathy.SIGNIFICANCE STATEMENT Somatodendritic localization of tau may be an early step leading to the neuronal degeneration in tauopathies. However, the mechanisms of the normal axonal distribution of tau and the mislocalization of pathological tau remain obscure. Our immunohistochemical and biochemical analyses demonstrated that the endogenous mouse tau is transiently expressed in neonatal brains, that exogenous human tau expressed corresponding to such tau expression profile can distribute into the axon, and that the constitutive expression of tau into adulthood (e.g., human tau in transgenic mice) results in abnormal somatodendritic localization. Thus, the expression profile of tau is tightly associated with the localization of tau, and the ectopic expression of tau in matured neurons may be involved in the pathogenesis of tauopathy.
Subject(s)
Brain Chemistry/physiology , Brain/cytology , Dendrites/physiology , Ectopic Gene Expression/genetics , tau Proteins/biosynthesis , Animals , Animals, Newborn , Axons/metabolism , Brain/growth & development , Female , Gene Knock-In Techniques , Humans , Immunohistochemistry , Male , Mice , Mice, Transgenic , Neurons/metabolism , Primary Cell Culture , Tauopathies/metabolismABSTRACT
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by accumulation of fragmented insoluble TDP-43 and loss of TDP-43 from the nucleus. Increased expression of exogenous TARDBP (encoding TDP-43) induces TDP-43 pathology and cytotoxicity, suggesting the involvement of aberrant expression of TDP-43 in the pathogenesis of ALS. In normal conditions, however, the amount of TDP-43 is tightly regulated by the autoregulatory mechanism involving alternative splicing of TARDBP mRNA. To investigate the influence of autoregulation dysfunction, we inhibited the splicing of cryptic intron 6 using antisense oligonucleotides in vivo. This inhibition doubled the Tardbp mRNA expression, increased the fragmented insoluble TDP-43, and reduced the number of motor neurons in the mouse spinal cord. In human induced pluripotent stem cell-derived neurons, the splicing inhibition of intron 6 increased TARDBP mRNA and decreased nuclear TDP-43. These non-genetically modified models exhibiting rise in the TARDBP mRNA levels suggest that TDP-43 autoregulation turbulence might be linked to the pathogenesis of ALS.
Subject(s)
DNA-Binding Proteins/metabolism , Homeostasis/physiology , Motor Neurons/metabolism , RNA, Messenger/metabolism , Spinal Cord/metabolism , Alternative Splicing/physiology , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/metabolism , Amyotrophic Lateral Sclerosis/pathology , Animals , DNA-Binding Proteins/genetics , Humans , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/pathology , Mice , Motor Neurons/pathology , RNA, Messenger/genetics , Spinal Cord/pathologyABSTRACT
BACKGROUND: Mycosis fungoides (MF) is the most common cutaneous T-cell lymphoma. Early-stage MF patches or plaques often resemble inflammatory skin disorders (ISDs), including psoriasis and atopic dermatitis. Cell adhesion molecule 1 gene (CADM1), which was initially identified as a tumor suppressor gene in human non-small cell lung cancer, has been reported as a diagnostic marker for adult T-cell leukemia/lymphoma. OBJECTIVE: We investigated CADM1 expression in MF neoplastic cells, especially during early stages, and evaluated its usefulness as a diagnostic marker for MF. METHODS: We conducted a retrospective study by using immunohistochemical staining and confirmed the expression of CADM1 in MF. In addition, we compared CADM1 messenger RNA expression in microdissected MF samples and ISD samples. RESULTS: In the overall study period, 55 of 58 MF samples (94.8 %) stained positive for CADM1. None of the 50 ISD samples showed positive reactivity (P < .0001). We found CADM1 messenger RNA expression in the intradermal lymphocytes of patients with MF but not in those of patients with an ISD. LIMITATIONS: We did not conduct a validation study for MF cases in other institutions. CONCLUSIONS: CADM1-positive cells can be identified in early stages with fewer infiltrating cells and may be useful as a diagnostic marker for early-stage MF.
Subject(s)
Biomarkers, Tumor/analysis , Cell Adhesion Molecule-1/analysis , Mycosis Fungoides/chemistry , Neoplasm Proteins/analysis , Adult , Aged , Aged, 80 and over , Cell Adhesion Molecule-1/biosynthesis , Cell Adhesion Molecule-1/genetics , Dermatitis/metabolism , Early Diagnosis , Female , Gene Expression Regulation, Neoplastic , Humans , Japan/epidemiology , Lymphocytes/metabolism , Lymphocytes/pathology , Middle Aged , Mycosis Fungoides/diagnosis , Mycosis Fungoides/genetics , Mycosis Fungoides/pathology , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Neoplasm Staging , RNA, Messenger/biosynthesis , RNA, Neoplasm/biosynthesis , Retrospective StudiesABSTRACT
Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder. In motor neurons of ALS, TAR DNA binding protein-43 (TDP-43), a nuclear protein encoded by TARDBP, is absent from the nucleus and forms cytoplasmic inclusions. TDP-43 auto-regulates the amount by regulating the TARDBP mRNA, which has three polyadenylation signals (PASs) and three additional alternative introns within the last exon. However, it is still unclear how the autoregulatory mechanism works and how the status of autoregulation in ALS motor neurons without nuclear TDP-43 is. Here we show that TDP-43 inhibits the selection of the most proximal PAS and induces splicing of multiple alternative introns in TARDBP mRNA to decrease the amount of cytoplasmic TARDBP mRNA by nonsense-mediated mRNA decay. When TDP-43 is depleted, the TARDBP mRNA uses the most proximal PAS and is increased in the cytoplasm. Finally, we have demonstrated that in ALS motor neurons-especially neurons with mislocalized TDP-43-the amount of TARDBP mRNA is increased in the cytoplasm. Our observations indicate that nuclear TDP-43 contributes to the autoregulation and suggests that the absence of nuclear TDP-43 induces an abnormal autoregulation and increases the amount of TARDBP mRNA. The vicious cycle might accelerate the disease progression of ALS.
Subject(s)
Amyotrophic Lateral Sclerosis/genetics , DNA-Binding Proteins/genetics , Motor Neurons/metabolism , RNA, Messenger/genetics , Spinal Cord/metabolism , Amyotrophic Lateral Sclerosis/metabolism , Amyotrophic Lateral Sclerosis/pathology , Cell Nucleus/metabolism , Cell Nucleus/ultrastructure , Cytoplasm/metabolism , Cytoplasm/ultrastructure , DNA-Binding Proteins/metabolism , Exons , Feedback, Physiological , Gene Expression Regulation , HEK293 Cells , Humans , Introns , Motor Neurons/pathology , RNA Stability , RNA, Messenger/metabolism , Signal Transduction , Spinal Cord/pathologyABSTRACT
BACKGROUND AND PURPOSE: The principal aim of this study was to clarify the clinical profile of pediatric patients with fruit allergies. SUBJECTS AND METHODS: We assigned 265 pediatric patients with fruit allergies who were receiving outpatient treatment at Aichi Children's Health and Medical Center and 32 who underwent kiwi or banana oral food chal- lenge (OFC) test to an oral symptom group or a systemic symptom group and retrospectively examined their clinical profiles and prognosis from their medical records. RESULTS: The most common cause of fruit allergy was kiwi (139 patients, 21% of all patients). Watermelon had the highest oral symptom induction rate (97%), whereas banana had the highest systemic reaction induction rate (46%). In the oral symptom group, the prevalence of hay fever was 54% and 66% of patients showed aller- gic reaction to multiple fruits. In contrast, in the systemic symptom group, the prevalence of pollen allergies was 24% and 77% of patients showed allergic reaction to a single fruit only. The results revealed that 65% of patients allergic to bananas had infant-onset allergy and seven among 20 patients were confirmed acquisition of tolerability by the OFC test. CONCLUSION: Our data suggest that there is a difference in the clinical profile and disease type between patients with oral symptoms and those with primarily systemic symptoms. It is important to differentiate between these patients for accurate diet guidance, safety management, and prognosis estimation.
Subject(s)
Food Hypersensitivity , Fruit/adverse effects , Adolescent , Child , Child, Preschool , Female , Food Hypersensitivity/diagnosis , Humans , Infant , Male , Young AdultABSTRACT
Disappearance of TAR-DNA-binding protein 43 kDa (TDP-43) from the nucleus contributes to the pathogenesis of amyotrophic lateral sclerosis (ALS), but the nuclear function of TDP-43 is not yet fully understood. TDP-43 associates with nuclear bodies including Gemini of coiled bodies (GEMs). GEMs contribute to the biogenesis of uridine-rich small nuclear RNA (U snRNA), a component of splicing machinery. The number of GEMs and a subset of U snRNAs decrease in spinal muscular atrophy, a lower motor neuron disease, suggesting that alteration of U snRNAs may also underlie the molecular pathogenesis of ALS. Here, we investigated the number of GEMs and U11/12-type small nuclear ribonucleoproteins (snRNP) by immunohistochemistry and the level of U snRNAs using real-time quantitative RT-PCR in ALS tissues. GEMs decreased in both TDP-43-depleted HeLa cells and spinal motor neurons in ALS patients. Levels of several U snRNAs decreased in TDP-43-depleted SH-SY5Y and U87-MG cells. The level of U12 snRNA was decreased in tissues affected by ALS (spinal cord, motor cortex and thalamus) but not in tissues unaffected by ALS (cerebellum, kidney and muscle). Immunohistochemical analysis revealed the decrease in U11/12-type snRNP in spinal motor neurons of ALS patients. These findings suggest that loss of TDP-43 function decreases the number of GEMs, which is followed by a disturbance of pre-mRNA splicing by the U11/U12 spliceosome in tissues affected by ALS.
Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Amyotrophic Lateral Sclerosis/pathology , DNA-Binding Proteins/genetics , Gemini of Coiled Bodies/metabolism , Motor Neurons/pathology , RNA, Small Nuclear/genetics , Ribonucleoproteins, Small Nuclear/metabolism , Amyotrophic Lateral Sclerosis/genetics , Cells, Cultured , DNA-Binding Proteins/metabolism , HeLa Cells , Humans , Motor Cortex/metabolism , Motor Cortex/pathology , Motor Neurons/metabolism , RNA Splicing , RNA, Small Nuclear/metabolism , Real-Time Polymerase Chain Reaction , Ribonucleoproteins, Small Nuclear/genetics , SMN Complex Proteins/genetics , SMN Complex Proteins/metabolism , Spinal Cord/metabolism , Spinal Cord/pathology , Thalamus/metabolism , Thalamus/pathologyABSTRACT
Although oxidoreductases are widely used in many applications, such as biosensors and biofuel cells, improvements in the function of existing oxidoreductases or the discovery of novel oxidoreductases with greater activities is desired. To increase the activity of oxidoreductases by directed evolution, a powerful screening technique for oxidoreductases is required. In this study, we demonstrate the utility of boron-doped diamond (BDD) microelectrodes for quantitative and potentially high-throughput measurement of the activity of NAD(P)-dependent oxidoreductases. We first confirmed that BDD microelectrodes can quantify the activity of low concentrations (10-100 pM) of glucose-6-phosphate dehydrogenase and alcohol dehydrogenase with a measuring time of 1 ms per sample. In addition, we found that poisoning of BDD microelectrodes can be repressed by optimizing the pH and by adding l-arginine to the enzyme solution as an antiaggregation agent. Finally, we fabricated a microfluidic device containing a BDD electrode for the first time and observed the elevation of the oxidation current of NADH with increasing flow rate. These results imply that the combination of a BDD microelectrode and microfluidics can be used for high-throughput screening of an oxidoreductase library containing a large number (>10(6)) of samples, each with a small (nanoliter) sample volume.
Subject(s)
Boron/chemistry , Diamond , Microelectrodes , Microfluidics/instrumentation , NADH, NADPH Oxidoreductases/analysis , High-Throughput Screening Assays , Hydrogen-Ion Concentration , Limit of DetectionABSTRACT
Age-related microangiopathy, also known as small vessel disease (SVD), causes damage to the brain, retina, liver, and kidney. Based on the DNA damage theory of aging, we reasoned that genomic instability may underlie an SVD caused by dominant C-terminal variants in TREX1, the most abundant 3'-5' DNA exonuclease in mammals. C-terminal TREX1 variants cause an adult-onset SVD known as retinal vasculopathy with cerebral leukoencephalopathy (RVCL or RVCL-S). In RVCL, an aberrant, C-terminally truncated TREX1 mislocalizes to the nucleus due to deletion of its ER-anchoring domain. Since RVCL pathology mimics that of radiation injury, we reasoned that nuclear TREX1 would cause DNA damage. Here, we show that RVCL-associated TREX1 variants trigger DNA damage in humans, mice, and Drosophila, and that cells expressing RVCL mutant TREX1 are more vulnerable to DNA damage induced by chemotherapy and cytokines that up-regulate TREX1, leading to depletion of TREX1-high cells in RVCL mice. RVCL-associated TREX1 mutants inhibit homology-directed repair (HDR), causing DNA deletions and vulnerablility to PARP inhibitors. In women with RVCL, we observe early-onset breast cancer, similar to patients with BRCA1/2 variants. Our results provide a mechanistic basis linking aberrant TREX1 activity to the DNA damage theory of aging, premature senescence, and microvascular disease.
Subject(s)
DNA Damage , Exodeoxyribonucleases , Phosphoproteins , Animals , Exodeoxyribonucleases/genetics , Exodeoxyribonucleases/metabolism , Humans , Phosphoproteins/genetics , Phosphoproteins/metabolism , Mice , Recombinational DNA Repair , Phenotype , Mutation , Drosophila/genetics , Aging/genetics , Aging/metabolism , Female , Drosophila melanogaster/genetics , Male , Retinal Diseases , Vascular Diseases , Hereditary Central Nervous System Demyelinating DiseasesABSTRACT
Cerebral small-vessel disease is a common disorder in elderly populations; however, its molecular basis is not well understood. We recently demonstrated that mutations in the high-temperature requirement A (HTRA) serine peptidase 1 (HTRA1) gene cause a hereditary cerebral small-vessel disease, cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). HTRA1 belongs to the HTRA protein family, whose members have dual activities as chaperones and serine proteases and also repress transforming growth factor-ß (TGF-ß) family signaling. We demonstrated that CARASIL-associated mutant HTRA1s decrease protease activity and fail to decrease TGF-ß family signaling. However, the precise molecular mechanism for decreasing the signaling remains unknown. Here we show that increased expression of ED-A fibronectin is limited to cerebral small arteries and is not observed in coronary, renal arterial or aortic walls in patients with CARASIL. Using a cell-mixing assay, we found that HTRA1 decreases TGF-ß1 signaling triggered by proTGF-ß1 in the intracellular space. HTRA1 binds and cleaves the pro-domain of proTGF-ß1 in the endoplasmic reticulum (ER), and cleaved proTGF-ß1 is degraded by ER-associated degradation. Consequently, the amount of mature TGF-ß1 is reduced. These results establish a novel mechanism for regulating the amount of TGF-ß1, specifically, the intracellular cleavage of proTGF-ß1 in the ER.
Subject(s)
Cerebrovascular Disorders/enzymology , Protein Precursors/metabolism , Protein Processing, Post-Translational , Serine Endopeptidases/metabolism , Transforming Growth Factor beta1/metabolism , Cell Line , Cerebrovascular Disorders/genetics , Cerebrovascular Disorders/metabolism , Endoplasmic Reticulum/enzymology , Endoplasmic Reticulum/genetics , Endoplasmic Reticulum/metabolism , High-Temperature Requirement A Serine Peptidase 1 , Humans , Protein Binding , Protein Precursors/genetics , Serine Endopeptidases/genetics , Signal Transduction , Transforming Growth Factor beta1/geneticsABSTRACT
BACKGROUND: A GGGGCC hexanucleotide repeat expansion in C9ORF72 occurs on a chromosome 9p21 locus that is linked with frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) in white populations. The diseases resulting from this expansion are referred to as 'c9FTD/ALS'. It has been suggested that c9FTD/ALS arose from a single founder. However, the existence of c9FTD/ALS in non-white populations has not been evaluated. RESULTS: We found two index familial ALS (FALS) patients with c9FTD/ALS in the Japanese population. The frequency of c9FTD/ALS was 3.4% (2/58 cases) in FALS. No patients with sporadic ALS (n=110) or control individuals (n=180) had the expansion. Neuropathological findings of an autopsy case were indistinguishable from those of white patients. Although the frequency of risk alleles identified in white subjects is low in Japanese, one patient had all 20 risk alleles and the other had all but one. The estimated haplotype indicated that the repeat expansion in these patients was located on the chromosome with the risk haplotype identified in white subjects. CONCLUSIONS: C9ORF72 repeat expansions were present in a Japanese cohort of ALS patients, but they were rare. Intriguingly, Japanese patients appear to carry the same risk haplotype identified in white populations.
Subject(s)
Amyotrophic Lateral Sclerosis/genetics , DNA Repeat Expansion , Proteins/genetics , Alleles , Amyotrophic Lateral Sclerosis/pathology , Autopsy , Brain/pathology , C9orf72 Protein , Fatal Outcome , Female , Genotype , Haplotypes , Humans , Inclusion Bodies/pathology , Japan , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Proto-Oncogene Proteins c-myc/genetics , Spinal Cord/pathology , TDP-43 Proteinopathies/pathologyABSTRACT
Stroke is the leading cause of severe long-term disability. Cell therapy has recently emerged as an approach to facilitate functional recovery in stroke. Although administration of peripheral blood mononuclear cells preconditioned by oxygen-glucose deprivation (OGD-PBMCs) has been shown to be a therapeutic strategy for ischemic stroke, the recovery mechanisms remain largely unknown. We hypothesised that cell-cell communications within PBMCs and between PBMCs and resident cells are necessary for a polarising protective phenotype. Here, we investigated the therapeutic mechanisms underlying the effects of OGD-PBMCs through the secretome. We compared levels of transcriptomes, cytokines, and exosomal microRNA in human PBMCs by RNA sequences, Luminex assay, flow cytometric analysis, and western blotting under normoxic and OGD conditions. We also performed microscopic analyses to assess the identification of remodelling factor-positive cells and evaluate angiogenesis, axonal outgrowth, and functional recovery by blinded examination by administration of OGD-PBMCs after ischemic stroke in Sprague-Dawley rats. We found that the therapeutic potential of OGD-PBMCs was mediated by a polarised protective state through decreased levels of exosomal miR-155-5p, and upregulation of vascular endothelial growth factor and a pluripotent stem cell marker stage-specific embryonic antigen-3 through the hypoxia-inducible factor-1α axis. After administration of OGD-PBMCs, microenvironment changes in resident microglia by the secretome promoted angiogenesis and axonal outgrowth, resulting in functional recovery after cerebral ischemia. Our findings revealed the mechanisms underlying the refinement of the neurovascular unit by secretome-mediated cell-cell communications through reduction of miR-155-5p from OGD-PBMCs, highlighting the therapeutic potential carrier of this approach against ischemic stroke.
Subject(s)
Ischemic Stroke , MicroRNAs , Stroke , Rats , Animals , Humans , Oxygen/metabolism , Glucose/metabolism , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/metabolism , Leukocytes, Mononuclear/metabolism , Rats, Sprague-Dawley , MicroRNAs/geneticsABSTRACT
Introduction: Allergic diseases affect both children and adults, but generation-specific prevalence rates are unclear. Methods: An online questionnaire was used from December 2021 to January 2022 to survey the prevalence of allergic diseases among staff and their families of designated allergic disease medical hospitals in Japan. In this study, bronchial asthma (BA), atopic dermatitis (AD), food allergies (FAs), allergic rhinitis (AR), allergic conjunctivitis (AC), metal allergies (MAs), and drug allergies (DAs) were the allergic diseases surveyed. Results: In total, 18,706 individuals were surveyed (median age, 36 years; quartile range, 18-50). Allergic disease was reported in 62.2% of respondents. Across all ages, prevalence rates were as follows: BA (14.7%), AD (15.6%), FAs (15.2%), AR (47.4%), AC (19.5%), MAs (1.9%), and DAs (4.6%). The prevalence of BA and AR was higher in male children, whereas that of FAs and AC was higher in adult females. The prevalence of MAs and DAs peaked during adulthood and predominated among females. Conclusions: Our results suggest that approximately two-thirds of the Japanese population may have an allergic disease, with AR being the most prevalent.
ABSTRACT
Neuregulin-1 (NRG1) signaling is thought to contribute to both neuronal development and schizophrenia neuropathology. Here, we describe the developmental effects of excessive peripheral NRG1 signals on synaptic activity and AMPA receptor expression of GABAergic interneurons in postnatal rodent neocortex. A core peptide common to all NRG1 variants (eNRG1) was subcutaneously administered to mouse pups. Injected eNRG1 penetrated the blood-brain barrier and activated ErbB4 NRG1 receptors in the neocortex, in which ErbB4 mRNA is predominantly expressed by parvalbumin-positive GABAergic interneurons. We prepared neocortical slices from juvenile mice that were receiving eNRG1 subchronically and recorded inhibitory synaptic activity from layer V pyramidal neurons. Postnatal eNRG1 treatment significantly enhanced polysynaptic IPSCs, although monosynaptic IPSCs were not affected. Examination of excitatory inputs to parvalbumin-containing GABAergic interneurons revealed that eNRG1 treatment significantly increased AMPA-triggered inward currents and the amplitudes and frequencies of miniature EPSCs (mEPSCs). Similar effects on mEPSCs were observed in mice treated with a soluble, full-length form of NRG1 type I. Consistent with the electrophysiologic data, expression of the AMPA receptor GluA1 (i.e., GluR1, GluRA) was upregulated in the postsynaptic density/cytoskeletal fraction prepared from eNRG1-treated mouse neocortices. Cortical GABAergic neurons cultured with eNRG1 exhibited a significant increase in surface GluA1 immunoreactivity at putative synaptic sites on their dendrites. These results indicate that NRG1 circulating in the periphery influences postnatal development of synaptic AMPA receptor expression in cortical GABAergic interneurons and may play a role in conditions characterized by GABA-associated neuropathologic processes.
Subject(s)
Interneurons/metabolism , Neocortex/growth & development , Neocortex/metabolism , Neuregulin-1/physiology , Peripheral Nervous System/physiology , Receptors, AMPA/biosynthesis , Receptors, AMPA/physiology , Signal Transduction/physiology , gamma-Aminobutyric Acid/physiology , Animals , Blotting, Western , Cells, Cultured , Electrophysiological Phenomena , ErbB Receptors/biosynthesis , ErbB Receptors/genetics , Excitatory Postsynaptic Potentials/physiology , Female , Gene Expression Regulation/genetics , Gene Expression Regulation/physiology , Immunohistochemistry , In Situ Hybridization , Interneurons/physiology , Mice , Mice, Inbred C57BL , Neocortex/embryology , Pregnancy , Pyramidal Cells/physiology , Rats , Rats, Sprague-Dawley , Receptor, ErbB-4Subject(s)
Allergens/adverse effects , Antigens, Plant/immunology , Food Hypersensitivity/blood , Gliadin/immunology , Immunoglobulin E/blood , Severity of Illness Index , Triticum/adverse effects , Allergens/immunology , Child, Preschool , Food Hypersensitivity/diagnosis , Humans , Probability , Triticum/immunologyABSTRACT
Activation of human endogenous retrovirus-K (HERV-K) is one of the proposed risk factors for amyotrophic lateral sclerosis (ALS). The HERV-K envelope protein has been reported to show neurotoxicity, and development of therapy with reverse transcriptase inhibitors is being investigated. On the other hand, some reports have failed to show HERV-K activation in ALS. In this study, we analyzed the expression of HERV-K mRNA in the motor cortex and spinal cord of 15 Japanese patients with sporadic ALS and 19 controls using reverse transcriptase droplet digital PCR. This revealed no significant increase of HERV-K expression in ALS-affected tissues, suggesting that the association between ALS and HERV-K remains questionable.
Subject(s)
Amyotrophic Lateral Sclerosis , Endogenous Retroviruses , Motor Cortex , Amyotrophic Lateral Sclerosis/genetics , Endogenous Retroviruses/genetics , Humans , Japan , RNA, MessengerABSTRACT
Maternal nutrition during pregnancy is one of the factors affecting the health of offspring. There are conflicting findings about the association between maternal vitamin D status and the development of allergic diseases in offspring. The purpose of this study is to evaluate the association between maternal vitamin D intake and the development of allergic diseases in offspring at 1 y of age. From an ongoing nationwide birth cohort study (the Japan Environment and Children's Study), we obtained information on maternal vitamin D intake, determined by a food frequency questionnaire, and parent-reported physician-diagnosed allergic diseases in offspring at 1 y of age. From the full dataset of 103,062 pregnancies, we analyzed complete data for 82,592 mother-offspring pairs. The prevalence of physician-diagnosed asthma, food allergy, and atopic dermatitis in the children was 2.5%, 6.6%, and 4.3%, respectively. The mean (± standard deviation) maternal vitamin D intake was 4.7±4.7 µg/d, which is much lower than the recommended amount in Japan (7 µg/d). After adjustment for various covariates, the odds ratios were significantly higher for asthma in the 2nd quintile and for food allergies in the 3rd and 4th quintiles compared with the 1st quintile. However, there were no clear associations between maternal vitamin D intake and the development of allergic diseases in offspring at 1 y of age, even in a large nation-wide cohort study. Protective effects of vitamin D supplementation remain unclear.