Kazuhiko Watabe

Kyorin University Faculty of Health Sciences, Japan · Department of Medical Technology (Neuropathology)

Formation of cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). Mutations in two genes encoding transactivation response (TAR) DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS), both of which are main constituents of cytoplasmic aggregates, have been identified in...

TAR DNA-binding protein 43 (TDP-43) is a main constituent of cytoplasmic aggregates in neuronal and glial cells in cases of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. We have previously demonstrated that adenovirus-transduced artificial TDP-43 cytoplasmic aggregates formation is enhanced by proteasome inhibition in vitro a...

Myelination is one of the most remarkable biological events in the neuron-glia interactions for the development of the mammalian nervous system. To elucidate molecular mechanisms of cell-to-cell interactions in myelin synthesis in vitro, establishment of the myelinating system in cocultures of continuous neuronal and glial cell lines are desirable....

Transactivation response DNA‐binding protein of 43 kDa (TDP‐43) is a major constituent of cytoplasmic aggregates in neuronal and glial cells in cases of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We have previously shown neuronal cytoplasmic aggregate formation induced by recombinant adenoviruses expressing hu...

The formation of misfolded protein aggregates is one of the pathological hallmarks of neurodegenerative diseases. We have previously demonstrated the cytoplasmic aggregate formation of adenovirally expressed transactivation response DNA‐binding protein of 43 kDa (TDP‐43), the main constituent of neuronal cytoplasmic aggregates in cases of amyotroph...

Hereditary transthyretin (TTR) amyloidosis (ATTRv) is characterized by TTR amyloid deposition in the peripheral nervous system. It remains unknown why variant TTR preferentially deposits in the peripheral nerves and dorsal root ganglia. We previously detected low levels of TTR expression in Schwann cells and established an immortalized Schwann cell...

Edaravone is a free-radical scavenger drug that was recently approved for the treatment of amyotrophic lateral sclerosis (ALS), a neurodegenerative disease. A pathological hallmark of ALS is the accumulation of ubiquitinated or phosphorylated aggregates of the 43-kDa transactive response DNA binding protein (TDP-43) within the cytoplasm of motor ne...

A vast body of evidence implicates increased oxidative stress and extracellular glutamate accumulation in the pathomechanism of sporadic amyotrophic lateral sclerosis (ALS). Cystine/glutamate antiporter (xCT) carries extracellular cystine uptake and intracellular glutamate release (cystine/glutamate exchange) in the presence of oxidative stress. Th...

Patients with transthyretin (TTR)‐type familial amyloid polyneuropathy (FAP) typically exhibit sensory dominant polyneuropathy and autonomic neuropathy. However, the molecular pathogenesis of the neuropathy remains unclear. In this study, we characterize the features of FAP TTR the substitution of lysine for glutamic acid at position 61 (E61K). Thi...

Previous studies on sporadic amyotrophic lateral sclerosis (SALS) demonstrated iron accumulation in the spinal cord and increased glutamate concentration in the cerebrospinal fluid. To clarify the relationship between the two phenomena, we first performed quantitative and morphological analyses of substances related to iron and glutamate metabolism...

Small integral membrane protein of the lysosome/late endosome (SIMPLE) is a 161-amino acid cellular protein that contains a characteristic C-terminal domain known as the SIMPLE-like domain (SLD), which is well conserved among species. Several studies have demonstrated that SIMPLE localizes to the trans-Golgi network (TGN), early endosomes, lysosome...

The increased glucose flux into the polyol pathway via aldose reductase ( AR ) is recognized as a major contributing factor for the pathogenesis of diabetic neuropathy, whereas little is known about the functional significance of AR in the peripheral nervous system. Spontaneously immortalized Schwann cell lines established from long‐term cultures o...

Lysosomal storage disorders are characterized by progressive accumulation of undigested macromolecules within the cell due to lysosomal dysfunction. 573C10 is a Schwann cell line derived from a mouse model of Niemann-Pick type C disease-1, NPC (−/−). Under serum-starved conditions, NPC (−/−) cells manifested impaired autophagy accompanied by an inc...

Proteasome inhibitiors accelerate the formation of TDP-43 aggregates. (a-c) Differentiated neurons were transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (DsR-(WT+CTF)) (red) followed by the treatment with DMSO (a), 0.5 μM MG-132 (b), or 1 μM lactacystin (c) for 24 hrs. Fixed cells were immunostained with phosho-TDP-43 (pS409/S410) (green) and TuJ...

Time-lapse video of 1464RTBB3pEGFP-derived neuronal cells (green) transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (red) (for Fig 4B). Images were captured by every 15 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

Time-lapse imaging of 1464RTBB3pSirius-derived neuronal cells transduced with AxEGFP-WT.TDP43 and AxEGFP-CTF.TDP43 (green) followed by 0.5 μM MG-132 (for Fig A in S4 Fig). Images were captured by every 15 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

Insoluble cytoplasmic aggregates are formed by EGFP-tagged TDP-43 adenoviruses in the presence of MG-132. (a) Time-lapse imaging of AxEGFP-WT.TDP43 and AxEGFP-CTF.TDP43 (EGFP-(WT+CTF); gray scale in top panel and green in bottom panels, respectively)-transduced 1464RTBB3pSirius neurons in the presence of MG-132. Cytoplasmic TDP-43 aggregates (arrow...

Time-lapse imaging of 1464RTBB3pEGFP-derived neuronal cells (green) transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (red) followed by 0.5 μM MG-132 (for Fig 4D). Images were captured by every 20 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

Time-lapse imaging of 1464RTBB3pSirius-derived neuronal cells transduced with AxEGFP-WT.TDP43 (green) and AxDsR-CTF.TDP43 (red) followed by 0.5 μM MG-132 (for Fig 5A). Images were captured by every 15 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

Combined DsRed-tagged WT and CTF TDP-43 transduction in the presence of MG-132 induces cytoplasmic aggregates in 1464R cells. (a-d) Differentiated neurons were transduced with AxDsR-WT.TDP43 (DsR-WT) (a,c) or AxDsR-CTF.TDP43 (DsR-CTF) (b,d) (red) followed by the treatment with DMSO (a,b) or 0.5 μM MG-132 (c,d). Fixed cells were immunostained with p...

EGFP-tagged WT and CTF TDP-43 expression shows similar properties to DsRed-tagged TDP-43 expression. (a-f) Differentiated neurons were transduced with AxEGFP-WT.TDP43 (EGFP-WT) (a,d), AxEGFP-CTF.TDP43 (EGFP-CTF) (b,e) or both (c,f) (green) followed by the treatment with DMSO (a-c) or 0.5 μM MG-132 (d-f). Fixed cells were immunostained with phosho-T...

Time-lapse video of 1464RTBB3pEGFP-derived neuronal cells (green) transduced with AxDsR-WT.TDP43 and AxDsR-CTF.TDP43 (red) followed by 0.5 μM MG-132 (for Fig 4C). Images were captured by every 15 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

Time-lapse imaging of 1464RTBB3pSirius-derived neuronal cells transduced with AxDsR-WT.TDP43 (red) and AxEGFP-CTF.TDP43 (green) followed by 0.5 μM MG-132 (for Fig B in S4 Fig). Images were captured by every 15 min. The obtained serial images were converted into movie with 5 frames/second. Scale bar = 15 μm. (MP4)

In the present study, we performed a comprehensive analysis to clarify the clinicopathological characteristics of patients with amyotrophic lateral sclerosis (ALS) that had progressed to result in a totally locked-in state (communication Stage V), in which all voluntary movements are lost and communication is impossible. In 11 patients, six had pho...

This study aimed to quantitatively analyze fasciculation potentials (FPs) and to investigate their relationship with muscle strength in amyotrophic lateral sclerosis (ALS). Fifty-one patients with sporadic ALS or progressive muscular atrophy (25 men, 26 women, mean age of 68 years) underwent needle EMG. We determined the duration, phase number, and...

Amiodarone hydrochloride (AMD), an anti-arrhythmic agent, has been shown to cause peripheral neuropathy; however, its pathogenesis remains unknown. We examined the toxic effects of AMD on an immortalized adult rat Schwann cell line, IFRS1, and cocultures of IFRS1 cells and adult rat dorsal root ganglion neurons or nerve growth factor-primed PC12 ce...

Introduction: This study aimed to determine the prognostic factors and the values that predict survival after percutaneous endoscopic gastrostomy (PEG) tube placement in patients with amyotrophic lateral sclerosis (ALS). Methods: We retrospectively analyzed the correlations for 97 consecutive patients with ALS between clinical parameters and sur...

Predictors of communication impairment in patients with amyotrophic lateral sclerosis (ALS) using tracheostomy-invasive ventilation (TIV) were investigated. Seventy-six ALS patients using TIV were enrolled and classified into three subgroups of communication ability: patients who could communicate with communication devices (Stage I), patients who...

Familial amyloidotic polyneuropathy (FAP) is one of the transthyretin (TTR) amyloidoses characterized by extracellular amyloid deposits and peripheral nerve involvement. Recently, we found significant expression of the TTR gene in Schwann cells of the peripheral nervous system. We hypothesized that local expression of variant TTR in Schwann cells m...

A spontaneously immortalized adult Fischer rat Schwann cell line IFRS1 retains the characteristic features of normal Schwann cells, and can be a useful tool for the study of diabetic neuropathy. In the present study, we examined the effects of high glucose and 3-deoxyglucosone (3-DG) on the viability and the protein expression of advanced glycation...

We describe a Japanese patient with familial amyotrophic lateral sclerosis (ALS) and a p.K510M mutation in the fused in sarcoma gene (FUS). The patient's condition was characterized clinically by an early onset and rapid progression. The patient eventually required mechanical ventilation and progressed to the totally locked-in state. Neuropathologi...

Previously we found that, after temporary cerebral ischemia, microvasculogenic secondary focal cerebral cortical ischemia occurred, caused by microvascular obstruction due to compression by swollen astrocytic end-feet, resulting in focal infarction. Herein, we examined whether mannitol infusion immediately after restoration of blood flow could prot...

We have established spontaneously immortalized Schwann cell lines from normal adult mice and rats, as well as murine disease models. One of the normal mouse cell lines, IMS32, possesses some biological properties of mature Schwann cells and high proliferative activities. The IMS32 cells have been utilized to investigate the action mechanisms of var...

We have established spontaneously immortalized Schwann cell lines from adult ICR mice [IMS32] and Fischer344 rats [IFRS1]. IMS32 cells display distinct Schwann cell phenotypes such as a spindle-shaped morphology and the expression of glial cell markers (e.g. S100, glial fibrillary acidic protein (GFAP), p75 low-affinity neurotrophin receptor (p75(N...

We investigated the relationship between fasciculation potentials (FPs) and survival in patients with ALS. In 85 ALS patients, we prospectively performed needle EMG in five to seven muscles of each patient. The shape of the detected FPs was analyzed by inspection, and FPs with >4 phases were judged as complex FPs. We analyzed the correlation betwee...

Molecular signaling of cardiac autonomic innervation is an unresolved issue. Here, we show that glial cell line-derived neurotrophic factor (GDNF) promotes cardiac sympathetic innervation in vitro and in vivo. In vitro, ventricular myocytes (VMs) and sympathetic neurons (SNs) isolated from neonatal rat ventricles and superior cervical ganglia were...

Galectin-1 (GAL-1), a member of a family of β-galactoside binding animal lectins, is predominantly expressed in isolectin B4 (IB4)-binding small non-peptidergic (glial cell line-derived neurotrophic factor (GDNF)-responsive) sensory neurons in the sections of adult rat dorsal root ganglia (DRG), but its functional role and the regulatory mechanisms...

The aim of myocardial tissue engineering is to repair or regenerate damaged myocardium with engineered cardiac tissue. However, this strategy has been hampered by lack of functional integration of grafts with native myocardium. Autonomic innervation may be crucial for grafts to function properly with host myocardium. In this study, we explored the...

Establishing stable coculture systems with neuronal and Schwann cell lines has been considered difficult, presumably because of their high proliferative activity and phenotypic differences from primary cultured cells. The present study is aimed at developing methods for myelin formation under coculture of the neural crest-derived pheochromocytoma c...

Abstract Astrocytes support neuronal functions by regulating the extracellular ion homeostasis and levels of neurotransmitters, and by providing fuel such as lactate to the neurons via their processes (APs). After two 10-min unilateral carotid occlusions with a 5-h interval in gerbils, we investigated maturing disseminated selective neuronal necros...

We established spontaneously immortalized Schwann cell lines from long-term cultures of adult Fischer 344 rat dorsal root ganglia (DRG) and peripheral nerves. One of these cell lines, designated immortalized Fischer rat Schwann cells 1 (IFRS1), showed spindle-shaped morphology; immunoreactivity for S100, p75 neurotrophin receptor (p75(NTR) ), glial...

Defect of metallothionein-III (MT-III) has been reported to be a contributor to the progression of amyotrophic lateral sclerosis (ALS). We explored the expression and effects of MT-III on the motor neurons of spinal cords of ALS model mice (G93A Cu/Zn superoxide dismutase (SOD-1) mutant-transgenic (Tg) mice) using a retrograde viral delivery system...

Amyotrophic lateral sclerosis (ALS) shows peculiar abnormalities of the autonomic nervous system, including sympathetic hyperactivity, which might result in sudden death. In general, the sympathetic hyperactivity could be caused by disruption of vagal inhibition. Our objective was to evaluate the vagus nerve morphometrically in autopsy cases of ALS...

We have established spontaneously immortalized Schwann cell lines from normal adult mice and rats and murine disease models. One of the normal mouse cell lines, IMS32, possesses some biological properties of mature Schwann cells and high proliferative activities. The IMS32 cells under hyperglycemic and/or hyperlipidemic conditions have been utilize...

Our objective was to describe cases of hyperosmolar hyperglycemic state (HHS) in advanced amyotrophic lateral sclerosis (ALS) patients and discuss its pathophysiology. Five ventilator-dependent patients with ALS, with no previous history of diabetes, showed development of marked hyperglycemia (plasma glucose levels of 755-1544 mg/dl) after precedin...

We examined the morphological changes of Golgi apparatus (GA) of the facial motor neurons in rats after facial nerve avulsion or axotomy. In rats after avulsion, the numbers of motor neurons showed reduction and fragmentation of GA, namely the organelle lost the normal network-like configuration which was replaced by numerous small disconnected ele...

Presenilin-1 (PSEN1) is a primary component of the gamma-secretase complex, and total levels of its holoprotein and endoproteolytic fragments are tightly regulated. We examined the effects of several types of endoplasmic reticulum (ER) stress on quantitative changes in the levels of PSEN1 mRNA, holoprotein, and fragments. The ER stress-inducing che...

We have constructed a chitosan nonwoven nanofiber mesh tube consisting of oriented fibers by the electrospinning method. The efficacy of oriented nanofibers on Schwann cell alignment and positive effect of this tube on peripheral nerve regeneration were confirmed. The physical properties of the chitosan nanofiber mesh sheets prepared by electrospin...

Calcium-independent group VIA phospholipase A(2) (iPLA(2)beta), encoded by PLA2G6, has been shown to be involved in various physiological and pathological processes, including immunity, cell death, and cell membrane homeostasis. Mutations in the PLA2G6 gene have been recently identified in patients with infantile neuroaxonal dystrophy (INAD). Subse...

Cultured Schwann cells under exposure to high glucose and methylglyoxal (MG) have been individually em-ployed for studying diabetic neuropathy; however, similarities and differences between these two culture models have not been studied. We investigated the effects of high glucose and MG on viability, polyol pathway activity, and expression of oxid...

Aims: Sporadic amyotrophic lateral sclerosis (ALS) is a progressive and invariably fatal disease involving the upper and lower motor neurones of adult humans. Among the neuropathological features of the disease, abnormalities in the protein-synthesizing system in motor neurones of the brainstem and spinal cord, such as a decrease of cytoplasmic RN...

The physiological roles of sonic hedgehog (Shh) have been intensively characterized in development of various organs. However, their functions in adult tissues have not been fully elucidated. We investigated the expression and the potential function of Shh in crush-injured adult rat sciatic nerves. The Shh expression was up-regulated in Schwann cel...

Motoneuron number and expression of cytoplasmic RNA and ribosomal RNA (rRNA) gene transcription activity in the facial nucleus were examined quantitatively and chronologically for up to 4 weeks in rats after facial nerve axotomy and avulsion in order to elucidate interrelationships in axonal changes. The right facial nerves of adult Fischer rats we...

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder characterized by selective loss of motoneurons leading to progressive paralysis and death by respiratory failure. Since the discovery of the mutation of Cu/Zn superoxide dismutase (SOD1) gene in patients with familial ALS, transgenic mice and rats expressing human mutan...

Two missense mutations (P123H and V70M) of beta-synuclein (beta-syn), the homologue of alpha-syn, have been recently identified in dementia with Lewy bodies. However, the mechanism through which these mutations influence the pathogenesis of dementia with Lewy bodies is unclear. To investigate the role of the beta-syn mutations in neurodegeneration,...

We have previously shown that gene therapy using Insulin-like growth factor (IGF)-I, glial cell line-derived neurotrophic factor (GDNF), and brain-derived neurotrophic factor (BDNF), or a combination of these trophic factors, is a treatment option for recurrent laryngeal nerve (RLN) palsy. However, there remain some difficulties preventing this opt...

Methylglyoxal (MG) is involved in the pathogenesis of diabetic complications via the formation of advanced glycation end products (AGEs) and reactive oxygen species (ROS). To clarify whether the antidiabetic drug metformin prevents Schwann cell damage induced by MG, we cultured mouse Schwann cells in the presence of MG and metformin. Cell apoptosis...

The surgical options for laryngeal paralysis only achieve static changes of vocal fold position. Laryngeal reinnervation procedures have had little impact on the return of dynamic laryngeal function. The development of a new treatment for laryngeal paralysis, aimed at the return of dynamic function and neurologic restoration and regeneration, is ne...

Peripheral neuropathy is one of the important manifestations of Fabry disease. Enzyme replacement therapy with presently available recombinant alpha-galactosidases does not always improve the Fabry neuropathy. But the reason has not been determined yet. We established a Schwann cell line from Fabry mice, characterized it, and then examined the upta...

Hepatoma-derived growth factor (HDGF) is a nuclear protein homologous to the high-mobility group B1 family of proteins. It is known to be released from cells and to act as a trophic factor for dividing cells. In this study HDGF was increased in spinal motor neurons of a mouse model of motor neuron degeneration, polyglutamine-tract-binding protein-1...