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      Ginkgo biloba extract increases neurite outgrowth and activates the Akt/mTOR pathway

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      1 , 2 , 1 , 2 , 1 , 2 , *
      PLoS ONE
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          Abstract

          Background

          Standardized Ginkgo biloba extract (GBE) has demonstrated efficacy in the cognitive functional neuropsychiatric symptoms of patients with Alzheimer’s disease (AD). With regard to its underlying molecular mode of action, first evidence was provided that GBE was able to modulate neuronal outgrowth in vitro, but the mechanisms underlying GBE effects on neuroplasticity remain unclear.

          Methodology/Principal findings

          In this study, we investigated the effect of GBE on neurite outgrowth using SH-SY5Y neuroblastoma cells in a 2D and 3D surface culture. The effects of the GBE LI1370 on neuroplasticity and neurite outgrowth were compared to those of nerve growth factor (NGF, 50 ng/ml) which was used as a positive control. We evaluated several parameters of neurite outgrowth such as the neurite number, total neurite length and extend of branching. Our findings showed that GBE (10 and 100 μg/ml) significantly increased neurite outgrowth in the 2D as well as 3D culture model after 3 days of treatment with a comparable effect than that NGF. The use of the 3D cell culture allowed us to better reproduce the in vivo neuronal microenvironment for the evaluation the neurite formation after GBE treatment. In addition, we assessed the effects of GBE on the Akt/mTOR pathway, which is known to promote neuroplasticity induced by nerve growth factors. We showed that GBE treatment induced an increase of phosphorylated IGF1R (Tyr1135/Tyr1136), Akt (Ser473), TSC2 (Ser939), mTOR (Ser2448), PTEN (Ser380) and GSK3β (Ser9).

          Conclusion

          Together, these findings indicate that GBE promotes neurite growth and activates the PI3K/Akt/mTOR pathway suggesting that this plant extract supports neuronal plasticity.

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          Most cited references35

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          TSC2 is phosphorylated and inhibited by Akt and suppresses mTOR signalling.

          Tuberous sclerosis (TSC) is an autosomal dominant disorder characterized by the formation of hamartomas in a wide range of human tissues. Mutation in either the TSC1 or TSC2 tumour suppressor gene is responsible for both the familial and sporadic forms of this disease. TSC1 and TSC2 proteins form a physical and functional complex in vivo. Here, we show that TSC1-TSC2 inhibits the p70 ribosomal protein S6 kinase 1 (an activator of translation) and activates the eukaryotic initiation factor 4E binding protein 1 (4E-BP1, an inhibitor of translational initiation). These functions of TSC1-TSC2 are mediated by inhibition of the mammalian target of rapamycin (mTOR). Furthermore, TSC2 is directly phosphorylated by Akt, which is involved in stimulating cell growth and is activated by growth stimulating signals, such as insulin. TSC2 is inactivated by Akt-dependent phosphorylation, which destabilizes TSC2 and disrupts its interaction with TSC1. Our data indicate a molecular mechanism for TSC2 in insulin signalling, tumour suppressor functions and in the inhibition of cell growth.
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            A Simple Role for BDNF in Learning and Memory?

            Since its discovery almost three decades ago, the secreted neurotrophin brain-derived neurotrophic factor (BDNF) has been firmly implicated in the differentiation and survival of neurons of the CNS. More recently, BDNF has also emerged as an important regulator of synaptogenesis and synaptic plasticity mechanisms underlying learning and memory in the adult CNS. In this review we will discuss our knowledge about the multiple intracellular signalling pathways activated by BDNF, and the role of this neurotrophin in long-term synaptic plasticity and memory formation as well as in synaptogenesis. We will show that maturation of BDNF, its cellular localization and its ability to regulate both excitatory and inhibitory synapses in the CNS may result in conflicting alterations in synaptic plasticity and memory formation. Lack of a precise knowledge about the mechanisms by which BDNF influences higher cognitive functions and complex behaviours may constitute a severe limitation in the possibility to devise BDNF-based therapeutics for human disorders of the CNS.
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              Mitochondria and neuroplasticity

              The production of neurons from neural progenitor cells, the growth of axons and dendrites and the formation and reorganization of synapses are examples of neuroplasticity. These processes are regulated by cell-autonomous and intercellular (paracrine and endocrine) programs that mediate responses of neural cells to environmental input. Mitochondria are highly mobile and move within and between subcellular compartments involved in neuroplasticity (synaptic terminals, dendrites, cell body and the axon). By generating energy (ATP and NAD+), and regulating subcellular Ca2+ and redox homoeostasis, mitochondria may play important roles in controlling fundamental processes in neuroplasticity, including neural differentiation, neurite outgrowth, neurotransmitter release and dendritic remodelling. Particularly intriguing is emerging data suggesting that mitochondria emit molecular signals (e.g. reactive oxygen species, proteins and lipid mediators) that can act locally or travel to distant targets including the nucleus. Disturbances in mitochondrial functions and signalling may play roles in impaired neuroplasticity and neuronal degeneration in Alzheimer's disease, Parkinson's disease, psychiatric disorders and stroke.
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                Author and article information

                Contributors
                Role: InvestigationRole: Writing – original draft
                Role: Writing – original draft
                Role: Supervision
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 December 2019
                2019
                : 14
                : 12
                : e0225761
                Affiliations
                [1 ] University of Basel, Neurobiology Lab for Brain Aging and Mental Health, Transfaculty Research Platform Molecular and Cognitive Neuroscience, Basel, Switzerland
                [2 ] Psychiatric University Clinics, University of Basel, Basel, Switzerland
                Lewis Katz School of Medicine at Temple University, UNITED STATES
                Author notes

                Competing Interests: AE received study grants from Vifor Pharma Switzerland. Moreover AE and AG received lecturer/consultant fees from Vifor Pharma Switzerland. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                Author information
                http://orcid.org/0000-0002-9341-3669
                Article
                PONE-D-19-18727
                10.1371/journal.pone.0225761
                6886765
                31790465
                52029555-2fbb-4b88-83aa-ee125218d2b1
                © 2019 Lejri et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 5 July 2019
                : 12 November 2019
                Page count
                Figures: 6, Tables: 0, Pages: 13
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100006484, Vifor Pharma;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100008947, Stiftung Synapsis - Alzheimer Forschung Schweiz AFS;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001711, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung;
                Award ID: 31003A_149728
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100008375, Universität Basel;
                Award Recipient :
                This study was partly supported by a principal investigator (AE) - initiated research grant supported by Vifor Pharma Switzerland, with regard to consumables and materials as well as GBE supply. Remaining funding was provided by grants from the Swiss National Science Foundation (SNF #31003A_149728, to AE) and Synapsis Foundation - Alzheimer Research Switzerland ARS to AG, and funding from the Transfaculty Research Platform, Molecular & Cognitive Neuroscience, University of Basel. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Neurons
                Neuronal Dendrites
                Neurites
                Biology and Life Sciences
                Neuroscience
                Cellular Neuroscience
                Neurons
                Neuronal Dendrites
                Neurites
                Research and Analysis Methods
                Biological Cultures
                Cell Cultures
                Cultured Tumor Cells
                Neuroblastoma Cells
                Biology and Life Sciences
                Neuroscience
                Cellular Neuroscience
                Neuronal Plasticity
                Research and Analysis Methods
                Biological Cultures
                Cell Cultures
                Biology and Life Sciences
                Biochemistry
                Bioenergetics
                Energy-Producing Organelles
                Mitochondria
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Energy-Producing Organelles
                Mitochondria
                Biology and Life Sciences
                Biochemistry
                Proteins
                Post-Translational Modification
                Phosphorylation
                Medicine and Health Sciences
                Endocrinology
                Endocrine Physiology
                Growth Factors
                Nerve Growth Factor
                Biology and Life Sciences
                Physiology
                Endocrine Physiology
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                Nerve Growth Factor
                Medicine and Health Sciences
                Physiology
                Endocrine Physiology
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                Biology and Life Sciences
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                Neurochemistry
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                Biology and Life Sciences
                Neuroscience
                Neurochemistry
                Neurochemicals
                Nerve Growth Factor
                Research and Analysis Methods
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                Confocal Microscopy
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