1.LRRK2 Inhibits FAK Activity by Promoting FERM-mediated Autoinhibition of FAK and Recruiting the Tyrosine Phosphatase, SHP-2.
Insup CHOI ; Ji won BYUN ; Sang Myun PARK ; Ilo JOU ; Eun Hye JOE
Experimental Neurobiology 2016;25(5):269-276
Mutation of leucine-rich repeat kinase 2 (LRRK2) causes an autosomal dominant and late-onset familial Parkinson's disease (PD). Recently, we reported that LRRK2 directly binds to and phosphorylates the threonine 474 (T474)-containing Thr-X-Arg(Lys) (TXR) motif of focal adhesion kinase (FAK), thereby inhibiting the phosphorylation of FAK at tyrosine (Y) 397 residue (pY397-FAK), which is a marker of its activation. Mechanistically, however, it remained unclear how T474-FAK phosphorylation suppressed FAK activation. Here, we report that T474-FAK phosphorylation could inhibit FAK activation via at least two different mechanisms. First, T474 phosphorylation appears to induce a conformational change of FAK, enabling its N-terminal FERM domain to autoinhibit Y397 phosphorylation. This is supported by the observation that the levels of pY397-FAK were increased by deletion of the FERM domain and/or mutation of the FERM domain to prevent its interaction with the kinase domain of FAK. Second, pT474-FAK appears to recruit SHP-2, which is a phosphatase responsible for dephosphorylating pY397-FAK. We found that mutation of T474 into glutamate (T474E-FAK) to mimic phosphorylation induced more strong interaction with SHP-2 than WT-FAK, and that pharmacological inhibition of SHP-2 with NSC-87877 rescued the level of pY397 in HEK293T cells. These results collectively show that LRRK2 suppresses FAK activation through diverse mechanisms that include the promotion of autoinhibition and/or the recruitment of phosphatases, such as SHP-2.
Focal Adhesion Protein-Tyrosine Kinases
;
Glutamic Acid
;
Parkinson Disease
;
Phosphoric Monoester Hydrolases
;
Phosphorylation
;
Phosphotransferases
;
Protein Tyrosine Phosphatase, Non-Receptor Type 11*
;
Threonine
;
Tyrosine*
2.Systematic Analysis of Translocator Protein 18 kDa (TSPO) Ligands on Toll-like Receptors-mediated Pro-inflammatory Responses in Microglia and Astrocytes.
Ji Won LEE ; Hyeri NAM ; Seong Woon YU
Experimental Neurobiology 2016;25(5):262-268
Translocator protein 18 kDa (TSPO) is a mitochondrial protein highly expressed on reactive microglia and astrocytes, and is considered as a biomarker for neurodegeneration and brain damage, especially neuroinflammation. Toll-like receptors (TLRs) are closely related with inflammatory responses of microglia and astrocytes and these signaling pathways regulate neuroinflammation. Previous reports have identified the anti-inflammatory effects of TSPO ligands, however study of their effects in relation to the TLR signaling was limited. Here, we investigated the effects of five representative TSPO ligands on microglia and astrocytes following activation by various TLR ligands. Our results show that TSPO ligands reduce the pro-inflammatory response elicited by the TLR ligands with more profound effects on microglia than astrocytes.
Astrocytes*
;
Brain
;
Ligands*
;
Microglia*
;
Mitochondrial Proteins
;
Toll-Like Receptors
3.Comparative Analysis of Protein Tyrosine Phosphatases Regulating Microglial Activation.
Gyun Jee SONG ; Jaehong KIM ; Jong Heon KIM ; Seungeun SONG ; Hana PARK ; Zhong Yin ZHANG ; Kyoungho SUK
Experimental Neurobiology 2016;25(5):252-261
Protein tyrosine phosphatases (PTPs) are key regulatory factors in inflammatory signaling pathways. Although PTPs have been extensively studied, little is known about their role in neuroinflammation. In the present study, we examined the expression of 6 different PTPs (PTP1B, TC-PTP, SHP2, MEG2, LYP, and RPTPβ) and their role in glial activation and neuroinflammation. All PTPs were expressed in brain and glia. The expression of PTP1B, SHP2, and LYP was enhanced in the inflamed brain. The expression of PTP1B, TC-PTP, and LYP was increased after treating microglia cells with lipopolysaccharide (LPS). To examine the role of PTPs in microglial activation and neuroinflammation, we used specific pharmacological inhibitors of PTPs. Inhibition of PTP1B, TC-PTP, SHP2, LYP, and RPTPβ suppressed nitric oxide production in LPS-treated microglial cells in a dose-dependent manner. Furthermore, intracerebroventricular injection of PTP1B, TC-PTP, SHP2, and RPTPβ inhibitors downregulated microglial activation in an LPS-induced neuroinflammation model. Our results indicate that multiple PTPs are involved in regulating microglial activation and neuroinflammation, with different expression patterns and specific functions. Thus, PTP inhibitors can be exploited for therapeutic modulation of microglial activation in neuroinflammatory diseases.
Brain
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Microglia
;
Neuroglia
;
Nitric Oxide
;
Protein Tyrosine Phosphatase, Non-Receptor Type 2
;
Protein Tyrosine Phosphatases*
4.Inflammation after Ischemic Stroke: The Role of Leukocytes and Glial Cells.
Jong Youl KIM ; Joohyun PARK ; Ji Young CHANG ; Sa Hyun KIM ; Jong Eun LEE
Experimental Neurobiology 2016;25(5):241-251
The immune response after stroke is known to play a major role in ischemic brain pathobiology. The inflammatory signals released by immune mediators activated by brain injury sets off a complex series of biochemical and molecular events which have been increasingly recognized as a key contributor to neuronal cell death. The primary immune mediators involved are glial cells and infiltrating leukocytes, including neutrophils, monocytes and lymphocyte. After ischemic stroke, activation of glial cells and subsequent release of pro- and anti-inflammatory signals are important for modulating both neuronal cell damage and wound healing. Infiltrated leukocytes release inflammatory mediators into the site of the lesion, thereby exacerbating brain injury. This review describes how the roles of glial cells and circulating leukocytes are a double-edged sword for neuroinflammation by focusing on their detrimental and protective effects in ischemic stroke. Here, we will focus on underlying characterize of glial cells and leukocytes under inflammation after ischemic stroke.
Brain
;
Brain Injuries
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Cell Death
;
Inflammation*
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Leukocytes*
;
Lymphocytes
;
Monocytes
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Neuroglia*
;
Neurons
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Neutrophils
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Stroke*
;
Wound Healing
5.Astrocytes and Microglia as Non-cell Autonomous Players in the Pathogenesis of ALS.
Junghee LEE ; Seung Jae HYEON ; Hyeonjoo IM ; Hyun RYU ; Yunha KIM ; Hoon RYU
Experimental Neurobiology 2016;25(5):233-240
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder that leads to a progressive muscle wasting and paralysis. The pathological phenotypes are featured by severe motor neuron death and glial activation in the lumbar spinal cord. Proposed ALS pathogenic mechanisms include glutamate cytotoxicity, inflammatory pathway, oxidative stress, and protein aggregation. However, the exact mechanisms of ALS pathogenesis are not fully understood yet. Recently, a growing body of evidence provides a novel insight on the importance of glial cells in relation to the motor neuronal damage via the non-cell autonomous pathway. Accordingly, the aim of the current paper is to overview the role of astrocytes and microglia in the pathogenesis of ALS and to better understand the disease mechanism of ALS.
Amyotrophic Lateral Sclerosis
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Astrocytes*
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Glutamic Acid
;
Microglia*
;
Motor Neurons
;
Neurodegenerative Diseases
;
Neuroglia
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Oxidative Stress
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Paralysis
;
Phenotype
;
Spinal Cord
6.Two-pore Domain Potassium Channels in Astrocytes.
Experimental Neurobiology 2016;25(5):222-232
Two-pore domain potassium (K(2P)) channels have a distinct structure and channel properties, and are involved in a background K⁺ current. The 15 members of the K(2P) channels are identified and classified into six subfamilies on the basis of their sequence similarities. The activity of the channels is dynamically regulated by various physical, chemical, and biological effectors. The channels are expressed in a wide variety of tissues in mammals in an isoform specific manner, and play various roles in many physiological and pathophysiological conditions. To function as channels, the K(2P) channels form dimers, and some isoforms form heterodimers that provide diversity in channel properties. In the brain, TWIK1, TREK1, TREK2, TRAAK, TASK1, and TASK3 are predominantly expressed in various regions, including the cerebral cortex, dentate gyrus, CA1-CA3, and granular layer of the cerebellum. TWIK1, TREK1, and TASK1 are highly expressed in astrocytes, where they play specific cellular roles. Astrocytes keep leak K⁺ conductance, called the passive conductance, which mainly involves TWIK1-TREK1 heterodimeric channel. TWIK1 and TREK1 also mediate glutamate release from astrocytes in an exocytosis-independent manner. The expression of TREK1 and TREK2 in astrocytes increases under ischemic conditions, that enhance neuroprotection from ischemia. Accumulated evidence has indicated that astrocytes, together with neurons, are involved in brain function, with the K(2P) channels playing critical role in these astrocytes.
Astrocytes*
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Brain
;
Cerebellum
;
Cerebral Cortex
;
Dentate Gyrus
;
Glutamic Acid
;
Ischemia
;
Mammals
;
Neurons
;
Neuroprotection
;
Potassium Channels*
;
Potassium*
;
Protein Isoforms
7.Optogenetic and Chemogenetic Approaches for Studying Astrocytes and Gliotransmitters.
Juwon BANG ; Hak Yeong KIM ; Hyosang LEE
Experimental Neurobiology 2016;25(5):205-221
The brain consists of heterogeneous populations of neuronal and non-neuronal cells. The revelation of their connections and interactions is fundamental to understanding normal brain functions as well as abnormal changes in pathological conditions. Optogenetics and chemogenetics have been developed to allow functional manipulations both in vitro and in vivo to examine causal relationships between cellular changes and functional outcomes. These techniques are based on genetically encoded effector molecules that respond exclusively to exogenous stimuli, such as a certain wavelength of light or a synthetic ligand. Activation of effector molecules provokes diverse intracellular changes, such as an influx or efflux of ions, depolarization or hyperpolarization of membranes, and activation of intracellular signaling cascades. Optogenetics and chemogenetics have been applied mainly to the study of neuronal circuits, but their use in studying non-neuronal cells has been gradually increasing. Here we introduce recent studies that have employed optogenetics and chemogenetics to reveal the function of astrocytes and gliotransmitters.
Astrocytes*
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Brain
;
In Vitro Techniques
;
Ions
;
Membranes
;
Neurons
;
Optogenetics*
8.Optogenetic Glia Manipulation: Possibilities and Future Prospects.
Woo Hyun CHO ; Ellane BARCELON ; Sung Joong LEE
Experimental Neurobiology 2016;25(5):197-204
Our brains are composed of two distinct cell types: neurons and glia. Emerging data from recent investigations show that glial cells, especially astrocytes and microglia, are able to regulate synaptic transmission and thus brain information processing. This suggests that, not only neuronal activity, but communication between neurons and glia also plays a key role in brain function. Thus, it is currently well known that the physiology and pathophysiology of brain function can only be completely understood by considering the interplay between neurons and glia. However, it has not yet been possible to dissect glial cell type-specific roles in higher brain functions in vivo. Meanwhile, the recent development of optogenetics techniques has allowed investigators to manipulate neural activity with unprecedented temporal and spatial precision. Recently, a series of studies suggested the possibility of applying this cutting-edge technique to manipulate glial cell activity. This review briefly discusses the feasibility of optogenetic glia manipulation, which may provide a technical innovation in elucidating the in vivo role of glial cells in complex higher brain functions.
Astrocytes
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Automatic Data Processing
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Brain
;
Humans
;
Microglia
;
Neuroglia*
;
Neurons
;
Optogenetics*
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Physiology
;
Research Personnel
;
Synapses
;
Synaptic Transmission
9.Role of Oxidative Stress in Parkinson's Disease.
Experimental Neurobiology 2013;22(1):11-17
Parkinson's disease (PD) is a progressive neurodegenerative movement disorder associated with a selective loss of the dopamine(DA)rgic neurons in the substantia nigra pars compacta and the degeneration of projecting nerve fibers in the striatum. Because there is currently no therapy that delays the neurodegenerative process, modification of the disease course by neuroprotective therapy is an important unmet clinical need. Toward this end, understanding cellular mechanisms that render the nigral neurons particularly vulnerable have been a subject of intensive research. Increasing evidence suggests that oxidative stress plays a major role. The metabolism of DA itself contributes to oxidative stress, resulting in modification of intracellular macromolecules whose functions are important for cell survival. Mitochondrial dysfunction and the consequent increase in reactive oxygen species also trigger a sequence of events that leads to cell demise. In addition, activated microglia produce nitric oxide and superoxide during neuroinflammatory responses, and this is aggravated by the molecules released by damaged DAergic neurons such as alpha-synuclein, neuromelanin and matrix metalloproteinase-3. Ways to reduce oxidative stress therefore can provide a therapeutic strategy. NAD(P)H:quinone reductase (NQO1) and other antioxidant enzymes, whose gene expression are commonly under the regulation of the transcription factor Nrf2, can serve as target proteins utilized toward development of disease-modifying therapy for PD.
alpha-Synuclein
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Cell Survival
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Dopamine
;
Gene Expression
;
Melanins
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Microglia
;
Movement Disorders
;
Nerve Fibers
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Neurons
;
Nitric Oxide
;
Oxidative Stress
;
Oxidoreductases
;
Parkinson Disease
;
Proteins
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Reactive Oxygen Species
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Substantia Nigra
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Superoxides
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Transcription Factors
10.Multi Target Neuroprotective and Neurorestorative Anti-Parkinson and Anti-Alzheimer Drugs Ladostigil and M30 Derived from Rasagiline.
Experimental Neurobiology 2013;22(1):1-10
Present anti-PD and -AD drugs have limited symptomatic activity and devoid of neuroprotective and neurorestorative property that is needed for disease modifying action. The complex pathology of PD and AD led us to develop several multi-target neuroprotective and neurorestorative drugs with several CNS targets with the ability for possible disease modifying activity. Employing the pharmacophore of our anti-parkinson drug rasagiline (Azilect, N-propagrgyl-1-R-aminoindan), we have developed a series of novel multi-functional neuroprotective drugs (A) [TV-3326 (N-propargyl-3R-aminoindan-5yl)-ethyl methylcarbamate)], with both cholinesterase-butyrylesterase and brain selective monoamine-oxidase (MAO) A/B inhibitory activities and (B) the iron chelator-radical scavenging-brain selective monoamine oxidase (MAO) A/B inhibitor and M30 possessing the neuroprotective and neurorescuing propargyl moiety of rasagiline, as potential treatment of AD, DLB and PD with dementia. Another series of multi-target drugs (M30, HLA-20 series) which are brain permeable iron chelators and potent selective brain MAO inhibitors were also developed. These series of drugs have the ability of regulating and processing amyloid precursor protein (APP) since APP and alpha-synuclein are metaloproteins (iron-regulated proteins), with an iron responsive element 5"UTR mRNA similar to transferring and ferritin. Ladostigil inhibits brain acetyl and butyrylcholinesterase in rats after oral doses. After chronic but not acute treatment, it inhibits MAO-A and -B in the brain. Ladostigil acts like an anti-depressant in the forced swim test in rats, indicating a potential for anti-depressant activity. Ladostigil prevents the destruction of nigrostriatal neurons induced by infusion of neurotoxin MPTP in mice. The propargylamine moiety of ladostigil confers neuroprotective activity against cytotoxicity induced by ischemia and peroxynitrite in cultured neuronal cells. The multi-target iron chelator M30 has all the properties of ladostigil and similar neuroprotective activity to ladostigil, but is not a ChE inhibitor. M30 has a neurorestorative activity in post-lesion of nigrostriatal dopamine neurons in MPTP, lacatcystin and 6-hydroxydopamine animal models of PD. The neurorestorative activity is related to the ability of the drug to activate hypoxia inducing factor (HIF) which induces the production of such neurotrophins as brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF) and erythropoietin as well as glia-derived neurotrophic factor (GDNF). The unique multiple actions of ladostigil and M30 make the potentially useful drugs for the treatment of dementia with Parkinsonian-like symptoms and depression.
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
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alpha-Synuclein
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Amyloid
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Animals
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Anoxia
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Brain
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Brain-Derived Neurotrophic Factor
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Butyrylcholinesterase
;
Chelating Agents
;
Dementia
;
Depression
;
Dopamine
;
Erythropoietin
;
Ferritins
;
Indans
;
Iron
;
Ischemia
;
Mice
;
Models, Animal
;
Monoamine Oxidase
;
Monoamine Oxidase Inhibitors
;
Nerve Growth Factors
;
Neurons
;
Neuroprotective Agents
;
Oxidopamine
;
Pargyline
;
Peroxynitrous Acid
;
Propylamines
;
Rats
;
RNA, Messenger
;
Vascular Endothelial Growth Factor A