Ayako Furukawa

Glyceraldehyde caused Alzheimer’s disease-like alterations in diagnostic marker levels in SH-SY5Y human neuroblastoma cells

Clinical evidence has implicated diabetes mellitus as one of the risk factors for the development and progression of Alzheimer’s disease (AD). However, the neurotoxic pathway activated due to abnormalities in glucose metabolism has not yet been identified in AD. In order to investigate the relationship between impaired cerebral glucose metabolism and the pathophysiology of AD, SH-SY5Y human neuroblastoma cells were exposed to glyceraldehyde (GA), an inhibitor of glycolysis. GA induced the production of GA-derived advanced glycation end-products (GA-AGEs) and cell apoptosis, glycolytic inhibition, decreases in the medium concentrations of diagnostic markers of AD, such as amyloid β 1-42 (Aβ42), and increases in tau phosphorylation. These results suggest that the production of GA-AGEs and/or inhibition of glycolysis induce AD-like alterations, and this model may be useful for examining the pathophysiology of AD.

A role of Heat Shock Protein 70 in Photoreceptor Cell Death: Potential as a Novel Therapeutic Target in Retinal Degeneration

Retinal degenerative diseases (RDs) such as retinitis pigmentosa (RP) are a genetically heterogeneous group of disorders characterized by night blindness and peripheral vision loss, which caused by the dysfunction and death of photoreceptor cells. Although many causative gene mutations have been reported, the final common end stage is photoreceptor cell death. Unfortunately, no effective treatments or therapeutic agents have been discovered. Heat shock protein 70 (HSP70) is highly conserved and has antiapoptotic activities. A few reports have shown that HSP70 plays a role in RDs. Thus, we focused on the role of HSP70 in photoreceptor cell death. Using the N‐methyl‐N‐nitrosourea (MNU)‐induced photoreceptor cell death model in mice, we could examine two stages of the novel cell death mechanism; the early stage, including HSP70 cleavage through protein carbonylation by production of reactive oxygen species, lipid peroxidation and Ca2+ influx/calpain activation, and the late stage of cathepsin and/or caspase activation. The upregulation of intact HSP70 expression by its inducer is likely to protect photoreceptor cells. In this review, we focus on the role of HSP70 and the novel cell death signaling process in RDs. We also describe candidate therapeutic agents for RDs.

Involvement of neuronal nitric oxide synthase in N-methyl-N-nitrosourea-induced retinal degeneration in mice

N-methyl-N-nitrosourea (MNU) is widely used to study the mechanism of retinal degenerative diseases (RDs) because of its selectivity of photoreceptor cell death. Many reports suggest that excessive nitric oxide (NO) plays a crucial role in neuronal cell death. We hypothesized that nitric oxide synthase (NOS)/NO are involved in photoreceptor cell death by MNU. We found that the levels of NO increased after MNU treatment. Furthermore, we demonstrated that neuronal NOS specific inhibitor attenuated photoreceptor cell death by MNU in mice. We believe that our findings might be a new target for the treatment of RDs.

Spontaneous occurrence of photoageing‐like phenotypes in the dorsal skin of old SAMP1 mice, an oxidative stress model

Skin photoageing is a complex, multifactorial process and both intrinsic and extrinsic factors may contribute to its pathogenesis. The ultraviolet‐irradiated hairless mouse has been used as an animal model for photoageing, but this model mimics only the ‘extrinsic’ aspects. Here, we show that skin from old SAMP1 mice, a model for higher oxidative stress and senescence acceleration, exhibited histological and gene expression changes similar to those in human photoaged skin without ultraviolet irradiation. These changes include an increase in elastic fibre and glycosaminoglycan histologically, an upregulation of several proinflammatory cytokines and matrix metalloproteinases, and an increase in lipid peroxide. We propose that SAMP1 mice are a spontaneous animal model for photoageing caused by an exaggerated intrinsic mechanism, namely, higher oxidative status. This mouse model is useful to explore the link between oxidative stress and photoageing, and to evaluate the efficacy of antioxidants.

S -Nitrosylation Regulates Cell Survival and Death in the Central Nervous System

Nitric oxide (NO), which is produced from nitric oxide synthase, is an important cell signaling molecule that is crucial for many physiological functions such as neuronal death, neuronal survival, synaptic plasticity, and vascular homeostasis. This diffusible gaseous compound functions as an effector or second messenger in many intercellular communications and/or cell signaling pathways. Protein S-nitrosylation is a posttranslational modification that involves the covalent attachment of an NO group to the thiol side chain of select cysteine residues on target proteins. This process is thought to be very important for the regulation of cell death, cell survival, and gene expression in the central nervous system (CNS). However, there have been few reports on the role of protein S-nitrosylation in CNS disorders. Here, we briefly review specific examples of S-nitrosylation, with particular emphasis on its functions in neuronal cell death and survival. An understanding of the role and mechanisms underlying the effects of protein S-nitrosylation on neurodegenerative/neuroprotective events may reveal a novel therapeutic strategy for rescuing neurons in neurodegenerative diseases.

Talaumidin Promotes Neurite Outgrowth of Staurosporine-Differentiated RGC-5 Cells Through PI3K/Akt-Dependent Pathways

Talaumidin, a tetrahydrofuran neolignan isolated from the root of Aristolochia arcuata, was an interesting small molecule with neurotrophic activity in the cultured neuron. Talaumidin can promote neurite outgrowth from neurons. However, the mechanism by which talaumidin exerts its neurotrophic actions on retinal neurons has not been elucidated to date. In this study, we describe that talaumidin has neurotrophic properties such as neurite outgrowth in neuroretinal cell line, RGC-5. Talaumidin promotes staurosporine-induced neurite outgrowth in RGC-5 cells. The neurite outgrowth effect of talaumidin was inhibited by phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002, but not by Erk inhibitor, PD98059. These data suggest that talaumidin promotes neurite outgrowth through PI3K/Akt pathway and that the potential of talaumidin serves as a promising lead compound for the treatment of retinal degenerative disorders.

Protein Carbonylation-Dependent Photoreceptor Cell Death Induced by N-Methyl-N-nitrosourea in Mice

Retinal degenerative diseases, such as retinitis pigmentosa, are characterized by night blindness and peripheral vision loss caused by the slowly progressive loss of photoreceptor cells. A comprehensive molecular mechanism of the photoreceptor cell death remains unclear. We previously reported that heat shock protein 70 (HSP70), which has a protective effect on neuronal cells, was cleaved by a calcium-dependent protease, calpain, in N-methyl-N-nitrosourea (MNU)-treated mice retina. Carbonylated HSP70 is much more vulnerable than noncarbonylated HSP70 to calpain cleavage. However, it was not known whether protein carbonylation occurs in MNU-treated mice retina. In this study, we clearly show protein carbonylation-dependent photoreceptor cell death induced by MNU in mice. Therefore, protein carbonylation and subsequent calpain-dependent cleavage of HSP70 are key events in MNU-mediated photoreceptor cell death. Our data provide a comprehensive molecular mechanism of the photoreceptor cell death.

Alternative Splicing for Activation of Coagulation Factor XIII-A in the Fish Retina After Optic Nerve Injury

Factor XIII-A (FXIII-A), which has become known as cellular transglutaminase, plays important roles in mediating cross-linking reactions in various tissues. FXIII-A acts as one of the regeneration molecules in the fish retina and optic nerve after optic nerve injury and becomes activated at the site of injury within a few hours. Previous research has shown that activated FXIII-A induces neurite outgrowth from injured retinal ganglion cells and supports elongation of the regenerating optic nerve. However, the activation mechanism of FXIII-A remains unknown. Furthermore, the injured tissues do not express thrombin, a known activator of plasma FXIII. Here, we investigated the mRNA expression of FXIII-A based on two different regions, one encoding the activation peptide and the other encoding the enzymatic active site. We found that expression of the region encoding the activation peptide was markedly suppressed compared with the region encoding the active site. An overexpression study with a short-type FXIII-A cDNA lacking the activation peptide revealed induction of long neurite outgrowth in fish retinal explant cultures compared with full-length FXIII-A cDNA. The present findings suggest that alternative splicing may occur in the FXIII-A gene, resulting in deletion of the region encoding the activation peptide and thus allowing direct production of activated FXIII-A protein in the fish retina and optic nerve after optic nerve injury.

HSP70 cleavage-induced photoreceptor cell death caused by N-methyl-N-nitrosourea

Retinal degenerative diseases (RDs) such as retinitis pigmentosa (RP) are characterized by slowly progressive photoreceptor cell death, but the molecular mechanism underlying RP remains unclear. Animal models for RP have led to a better understanding of the disease pathological mechanisms, yet it remains difficult to identify an appropriate genetic model for RDs in general because there are many causative genes (Rossmiller et al., 2012).

A potential of novel therapeutic approach by heat shock protein 70 expression in retinitis pigmentosa

要約:網膜色素変性症(retinitis pigmentosa:RP)は, わが国でも患者数が多いが,未だに有効な治療薬や治 療法が確立されておらず,2015 年から指定難病に定め られた.今後,超高齢化社会を迎える日本ではさらな る患者数の増加が見込まれることから,RPの治療法 確立が急務である.RPの最終病態は網膜視細胞の脱 落である.この病態形成に関わる細胞内イベントとし て活性酸素,細胞内カルシウム,カルシウム依存性プ ロテアーゼ,アポトーシス関連分子がそれぞれ報告さ れている.しかしながら,これらの報告は RPの病態 形成の一部を解明したものであり,相互の関連性が明 らかでないことから,その全体像は未だに不明である. 最近,70 kDa熱ショックタンパク質(HSP70)がアポ トーシスを抑制し,細胞保護機能を示すことが注目さ れている.しかし RPにおけるHSP70 の役割は解明さ れていない.近年,我々は HSP70 が活性酸素により カルボニル化修飾され,カルシウム依存性カルパイン により切断されることで視細胞死を引き起こすことを 初めて明らかにした.さらに HSP70 発現誘導薬を用 いて HSP70 を補うことで細胞死を抑制できることを 解明した.本総説において,我々は RPの新規治療薬 開発の標的として HSP70 発現誘導薬を提唱する.さ らにドラッグリポジショニングの観点から,HSP70 発 現誘導作用をもつ既存医薬品による新規 RP治療薬と しての可能性について述べる.