Atypical, non-standard functions of the microtubule associated Tau protein.

Since the discovery of the microtubule-associated protein Tau (MAPT) over 40 years ago, most studies have focused on Tau's role in microtubule stability and regulation, as well as on the neuropathological consequences of Tau hyperphosphorylation and aggregation in Alzheimer's disease (AD) brains. In recent years, however, research efforts identified new interaction partners and different sub-cellular localizations for Tau suggesting additional roles beyond its standard function as microtubule regulating protein. Moreover, despite the increasing research focus on AD over the last decades, Tau was only recently considered as a promising therapeutic target for the treatment and prevention of AD as well as for neurological pathologies beyond AD e.g. epilepsy, excitotoxicity, and environmental stress. This review will focus on atypical, non-standard roles of Tau on neuronal function and dysfunction in AD and other neurological pathologies providing novel insights about neuroplastic and neuropathological implications of Tau in both the central and the peripheral nervous system

Regulation of EB1/3 proteins by classical MAPs in neurons

Microtubules (MTs) are key cytoskeletal elements in developing and mature neurons. MT reorganization underlies the morphological changes that occur during neuronal development. Furthermore, MTs contribute to the maintenance of neuronal architecture, enable intracellular transport and act as scaffolds for signaling molecules. Thus, a fine-tuned regulation of MT dynamics and stability is crucial for the correct differentiation and functioning of neurons. Different types of proteins contribute to the regulation of the MT state, such as plus-end tracking proteins (+TIPs), which interact with the plus-ends of growing microtubules, and classical microtubule-associated proteins (MAPs), which bind along the microtubule lattice. Recent evidence indicates that MAPs interplay with End Binding Proteins (EBs), the core +TIPs, in neuronal cells. This might contribute to the orchestrated regulation of MT dynamics in neurons. In this mini-review article, we address recent research on the neuronal crosstalk between EBs and classical MAPs and speculate on its possible functional relevance. © 2014 Landes Bioscience.CSIC and CIBERNED (Madrid) and is currently supported and funded by the IMBRAIN project. The authors would like to acknowledge the IMBRAIN project (FP7-REGPOT-2012-CT2012–31637-IMBRAIN), funded under the 7th Framework Programme (Capacities). Our work was supported by grants of the Spanish Research Council (CSIC), Plan Nacional (MINECO), CIBERNED and Comunidad de Madri

GSK-3 and Tau: A Key Duet in Alzheimer's Disease

Glycogen synthase kinase-3 (GSK-3) is a ubiquitously expressed serine/threonine kinase with a plethora of substrates. As a modulator of several cellular processes, GSK-3 has a central position in cell metabolism and signaling, with important roles both in physiological and pathological conditions. GSK-3 has been associated with a number of human disorders, such as neurodegenerative diseases including Alzheimer's disease (AD). GSK-3 contributes to the hyperphosphorylation of tau protein, the main component of neurofibrillary tangles (NFTs), one of the hallmarks of AD. GSK-3 is further involved in the regulation of different neuronal processes that are dysregulated during AD pathogenesis, such as the generation of amyloid-β (Aβ) peptide or Aβ-induced cell death, axonal transport, cholinergic function, and adult neurogenesis or synaptic function. In this review, we will summarize recent data about GSK-3 involvement in these processes contributing to AD pathology, mostly focusing on the crucial interplay between GSK-3 and tau protein. We further discuss the current development of potential AD therapies targeting GSK-3 or GSK-3-phosphorylated tau.Spanish Ministry of Economy and Competitiveness (PGC-2018-09177-B-100

Regulation of neuronal cytoskeleton by lysophosphatidic acid: role of GSK-3

Neurite retraction is a crucial process during nervous system development and neurodegeneration. This process implies reorganization of the neuronal cytoskeleton. Some bioactive lipids such as lysophosphatidic acid (LPA) induce neurite retraction. The reorganization of the actin cytoskeleton during neurite retraction is one of the best-characterized effects of LPA. However, less information is available regarding the reorganization of the microtubule (MT) network in response to LPA in neuronal cells. Here, we first give an overview of the roles of cytoskeleton during neurite outgrowth, and subsequently, we review some of the data from different laboratories concerning LPA-induced cytoskeletal rearrangement in neuronal cells. We also summarize our own recent results about modifications of MTs during LPA-induced neurite retraction. We have shown that LPA induces changes in tubulin pools and increases in the phosphorylation levels of microtubule-associated proteins (MAPs), such as Tau. Tau hyperphosphorylation in response to LPA is mediated by the activation of glycogen synthase kinase-3 (GSK-3). The upregulation of GSK-3 activity by LPA seems to be a general process as it occurs in diverse neuronal cells of different species in correlation with the neurite retraction process.Our research was supported by institutional grants from Spanish CICYT and Ramón Areces Foundation.Peer reviewe

Glycogen synthase kinase-3 is activated in neuronal cells by Gα12 and Gα13 by rho-independent and rho-dependent mechanisms

Glycogen synthase kinase-3 (GSK-3) was generally considered a constitutively active enzyme, only regulated by inhibition. Here we describe that GSK-3 is activated by lysophosphatidic acid (LPA) during neurite retraction in rat cerebellar granule neurons. GSK-3 activation correlates with an increase in GSK-3 tyrosine phosphorylation. In addition, LPA induces a GSK-3-mediated hyperphosphorylation of the microtubule-associated protein tau. Inhibition of GSK-3 by lithium partially blocks neurite retraction, indicating that GSK-3 activation is important but not essential for the neurite retraction progress. GSK-3 activation by LPA in cerebellar granule neurons is neither downstream of Gαi nor downstream of Gαq/phospholipase C, suggesting that it is downstream of Gα12/13. Overexpression of constitutively active Gα12 (Gα12QL) and Gα13(Gα13QL) in Neuro2a cells induces upregulation of GSK-3 activity. Furthermore, overexpression of constitutively active RhoA (RhoAV14) also activates GSK-3 However, the activation of GSK-3 by Gα13 is blocked by coexpression with C3 transferase, whereas C3 does not block GSK-3 activation by Gα12. Thus, we demonstrate that GSK-3 is activated by both Gα12 and Gα13 in neuronal cells. However, GSK-3 activation by Gα13 is Rho-mediated, whereas GSK-3 activation by Gα12 is Rho-independent. The results presented here imply the existence of a previously unknown mechanism of GSK-3 activation by Gα12/13 subunits.This research was supported by grants from Spanish Comision Interministerial de Ciencia y Tecnologia and an institutional grant from Ramon Areces FoundationPeer reviewe

Glycogen Synthase Kinase-3 Is Activated in Neuronal Cells by Gα 12

Glycogen synthase kinase-3 (GSK-3) was generally considered a constitutively active enzyme, only regulated by inhibition. Here we describe that GSK-3 is activated by lysophosphatidic acid (LPA) during neurite retraction in rat cerebellar granule neurons. GSK-3 activation correlates with an increase in GSK-3 tyrosine phosphorylation. In addition, LPA induces a GSK-3-mediated hyperphosphorylation of the microtubule-associated protein tau. Inhibition of GSK-3 by lithium partially blocks neurite retraction, indicating that GSK-3 activation is important but not essential for the neurite retraction progress. GSK-3 activation by LPA in cerebellar granule neurons is neither downstream of Gαi nor downstream of Gαq/phospholipase C, suggesting that it is downstream of Gα12/13. Overexpression of constitutively active Gα12 (Gα12QL) and Gα13(Gα13QL) in Neuro2a cells induces upregulation of GSK-3 activity. Furthermore, overexpression of constitutively active RhoA (RhoAV14) also activates GSK-3 However, the activation of GSK-3 by Gα13 is blocked by coexpression with C3 transferase, whereas C3 does not block GSK-3 activation by Gα12. Thus, we demonstrate that GSK-3 is activated by both Gα12 and Gα13 in neuronal cells. However, GSK-3 activation by Gα13 is Rho-mediated, whereas GSK-3 activation by Gα12 is Rho-independent. The results presented here imply the existence of a previously unknown mechanism of GSK-3 activation by Gα12/13 subunits.This research was supported by grants from Spanish Comision Interministerial de Ciencia y Tecnologia and an institutional grant from Ramon Areces FoundationPeer reviewe

MAP1B regulates microtubule dynamics by sequestering EB1/3 in the cytosol of developing neuronal cells

MAP1B, a structural microtubule (MT)-associated protein highly expressed in developing neurons, plays a key role in neurite and axon extension. However, not all molecular mechanisms by which MAP1B controls MT dynamics during these processes have been revealed. Here, we show that MAP1B interacts directly with EB1 and EB3 (EBs), two core 'microtubule plus-end tracking proteins' (+TIPs), and sequesters them in the cytosol of developing neuronal cells. MAP1B overexpression reduces EBs binding to plus-ends, whereas MAP1B downregulation increases binding of EBs to MTs. These alterations in EBs behaviour lead to changes in MT dynamics, in particular overstabilization and looping, in growth cones of MAP1B-deficient neurons. This contributes to growth cone remodelling and a delay in axon outgrowth. Together, our findings define a new and crucial role of MAP1B as a direct regulator of EBs function and MT dynamics during neurite and axon extension. Our data provide a new layer of MT regulation: a classical MAP, which binds to the MT lattice and not to the end, controls effective concentration of core +TIPs thereby regulating MTs at their plus-ends. © 2013 European Molecular Biology Organization.Spanish Research Ministry (SAF2006-02424 and SAF2011-24841); CIBERNED; Comunidad de Madrid (S2010/BMD2331); Spanish Research Council (CSIC)Peer Reviewe

The inhibition of phosphatidylinositol-3-kinase induces neurite retraction and activates GSK3

It has been extensively described that neuronal differentiation involves the signalling through neurotrophin receptors to a Ras-dependent mitogen-activated protein kinase (MAPK) cascade. However, signalling pathways from other neuritogenic factors have not been well established. It has been reported that cAMP may activate protein kinase (PKA), and it has been shown that PKA-mediated stimulation of MAPK pathway regulates not only neuritogenesis but also survival. However, extracellular regulated kinases (ERKs) mediated pathways are not sufficient to explain all the processes which occur in neuronal differentiation. Our present data show that: in cAMP-mediated neuritogenesis, using the SH-SY5Y human neuroblastoma cell line, there exists a link between the activation of PKA and stimulation of phosphatidylinositol 3-kinase (PI3K). Both kinase activities are essential to the initial elongation steps. Surprisingly, this neuritogenic process appears to be independent of ERKs. While the activity of PI3K is essential for elongation and maintenance of neurites, its inhibition causes retraction. In this neurite retraction process, GSK3 is activated. Using both a pharmacological approach and gene transfer of a dominant negative form of GSK3, we conclude that this induced retraction is a GSK3-dependent process which in turn appears to be a common target for transduction pathways involved in lysophosphatidic acid-mediated and PI3K-mediated neurite retraction.This work was supported by grants DGCYT (PB93–0155 and PB94–0040), Fundacion La Caixa, and from an Institutional grant Fundacion Areces. Dr F. Lim was supported during 1999 by a grant from Plan Nacional BIO98–0895.Peer reviewe

Tau regulates the localization and function of End-binding proteins 1 and 3 in developing neuronal cells

The axonal microtubule-associated protein tau is a well-known regulator of microtubule stability in neurons. However, the putative interplay between tau and End-binding proteins 1 and 3 (EB1/3), the core microtubule plus-end tracking proteins, has not been elucidated yet. Here, we show that a cross-talk between tau and EB1/3 exists in developing neuronal cells. Tau and EBs partially colocalize at extending neurites of N1E- 115 neuroblastoma cells and axons of primary hippocampal neurons, as shown by confocal immunofluorescence analyses. Tau down-regulation leads to a reduction of EB1/3 comet length, as observed in shRNA-stably depleted neuroblastoma cells and TAU-/- neurons. EB1/3 localization depends on the expression levels and localization of tau protein. Overexpression of tau at high levels induces EBs relocalization to microtubule bundles at extending neurites of N1E-115 cells. In differentiating primary neurons, tau is required for the proper accumulation of EBs at stretches of microtubule bundles at the medial and distal regions of the axon. Tau interacts with EB proteins, as shown by immunoprecipitation in different nonneuronal and neuronal cells and in whole brain lysates. A tau/ EB1 direct interaction was corroborated by in vitro pull-down assays. Fluorescence recovery after photobleaching assays performed in neuroblastoma cells confirmed that tau modulates EB3 cellular mobility. In summary, we provide evidence of a new function of tau as a direct regulator of EB proteins in developing neuronal cells. This cross-talk between a classical microtubule-associated protein and a core microtubule plusend tracking protein may contribute to the fine-tuned regulation of microtubule dynamics and stability during neuronal differentiation.Fil: Sayas, Carmen Laura. Universidad Autonoma de Madrid. Centro de Biología Molecular; EspañaFil: Tortosa, Elena. Universidad Autonoma de Madrid. Centro de Biología Molecular; EspañaFil: Bollati, Flavia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Farmacología Experimental de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Farmacología Experimental de Córdoba; ArgentinaFil: Ramírez Ríos, Sacnicte. Grenoble Institut Des Neurosciences-inserm U836; FranciaFil: Arnal, Isabelle. Grenoble Institut Des Neurosciences-inserm U836 ; FranciaFil: Avila, Jesús. Universidad Autonoma de Madrid. Centro de Biología Molecular; Españ