SUMOylation regulates nucleo-cytoplasmic shuttling of Elk-1

The transcription factor Elk-1 is a nuclear target of mitogen-activated protein kinases and regulates immediate early gene activation by extracellular signals. We show that Elk-1 is also conjugated to SUMO on either lysines 230, 249, or 254. Mutation of all three sites is necessary to fully block SUMOylation in vitro and in vivo. This Elk-1 mutant, Elk-1(3R), shuttles more rapidly to nuclei of Balb/C cells fused to transfected HeLa cells. Coexpression of SUMO-1 or -2 strongly reduces shuttling by Elk-1 without affecting that of Elk-1(3R), indicating that SUMOylation regulates nuclear retention of Elk-1. Accordingly, overexpression of Elk-1(3R) in PC12 cells, where cytoplasmic relocalization of Elk-1 has been linked to differentiation, enhances neurite extension relative to Elk-1. The effect of Elk-1, but not of the 3R mutant, was blocked upon cotransfection with SUMO-1 or -2 and enhanced by coexpression with mutant Ubc-9. Thus, SUMO conjugation is a novel regulator of Elk-1 function through the control of its nuclear-cytoplasmic shuttling

Neuron-gated silicon nanowire field effect transistors to follow single spike propagation within neuronal network

International audienceSilicon nanowire field effect transistors SiNW-FETs provide a local probe for sensing neuronal activity at the subcellular scale, thanks to their nanometer size and ultrahigh sensitivity. The combination with micro-patterning or microfluidic techniques to build model neurons networks above SiNW arrays could allow monitoring spike propagation and tailor specific stimulations, being useful to investigate network communications at multiple scales, such as plasticity or computing processes. This versatile device could be useful in many research areas, including diagnosis, prosthesis, and health security. Using top-down silicon nanowires-based array, we show here the ability to record electrical signals from matured neurons with top-down silicon nanowires, such as local field potential and unitary spike within ex-vivo preparations and hippocampal neurons grown on chip respectively. Furthermore, we demonstrate the ability to guide neurites above the sensors array during 3 weeks of cultures and follow propagation of spikes along cells. Silicon nanowire field effect transistors are obtained by top-down approach with CMOS compatible technology, showing the possibility to implement them at manufacturing level. These results confirm further the potentiality of the approach to follow spike propagation over large distances and at precise location along neuronal cells, by providing a multiscale addressing at the nano and mesoscales

Sensing ion channel in neuron networks with graphene field effect transistors

Graphene, the atomically-thin honeycomb carbon lattice, is a highly conducting 2D material whose exposed electronic structure offers an ideal platform for chemical and biological sensing. Its biocompatible, flexible and chemically inert nature associated with the lack of dangling bonds, offers novel perspectives for direct interfacing with biological molecules. Combined with its exceptional electronic and optical properties, this promotes graphene as a unique platform for bioelectronics. Among the successful bio-integrations of graphene, the detection of action potentials in numerous electrogenic cells including neurons has paved the road for the high spatio-temporal and wide-field mapping of neuronal activity. Ultimate resolution of sensing ion channel activity can be achieved with neural interfaces, and it was shown that macroscale electrodes can record extracellular current of individual ion channels in model systems, by charging the quantum capacitance of large graphene monolayer (mm2). Here, we show the field effect detection of ion channel activity within neuron networks, cultured during several weeks above graphene transistor arrays. Dependences upon drugs, reference potential gating and device geometry confirm the field effect detection of individual ion channel and suggest a significant contribution of grain boundaries, which provide highly sensitive nanoscale-sized sensing sites. Our theoretical analysis and simulations demonstrate that the ion gating of a single grain boundary in liquid affects the electronic transmission of the whole transistor channel, resulting in significant conductance variations. Monitoring the ion channels activity is of great interest as most of neurodegenerative diseases relied on channelopathies, which rely on ion channel abnormal activity. Thus, such highly sensitive and biocompatible neuro-electronics which open the way to FET detection at the sub-cell precision should be useful for a wide range of fundamental and applied research areas, including brain-on-chip, pharmacology, and in vivo monitoring or diagnosis.The authors gratefully acknowledge financial support from the University Joseph Fourrier (SMINGUE project), from la Région Rhône-Alpes (COOPERA project), and from the french Agence Nationale de la Recherche under the projects ANR-10-LABX-51-01 (Labex LANEF du Programme d'Investissements d'Avenir) the Lab Alliances on Nanosciences—Energies for the Future. ICN2 is supported by the CERCA programme/Generalitat de Catalunya. And the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (Project No. FIS2015-67767-P MINECO/FEDER), MINECO (Grant No. SEV-2013-0295) and funded by the CERCA Programme/Generalitat de Catalunya. The authors acknowledge the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (Project No. FIS2015-67767-P MINECO/FEDER).Peer reviewe