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000829528 0247_ $$2doi$$a10.3389/conf.fnins.2016.93.00080
000829528 0247_ $$2ISSN$$a1662-453X
000829528 0247_ $$2ISSN$$a1662-4548
000829528 037__ $$aFZJ-2017-03213
000829528 082__ $$a610
000829528 1001_ $$0P:(DE-Juel1)162348$$aTihaa, Irina$$b0$$eCorresponding author
000829528 1112_ $$a10th International Meeting on Substrate-Integrated Electrode Arrays$$cReutlingen$$d2016-06-28 - 2016-07-01$$wGermany
000829528 245__ $$aNeuronal guiding: Designing In Vitro Networks On MEA
000829528 260__ $$aLausanne$$bFrontiers Research Foundation$$c2016
000829528 300__ $$a1
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000829528 3367_ $$0PUB:(DE-HGF)8$$2PUB:(DE-HGF)$$aContribution to a conference proceedings$$bcontrib$$mcontrib$$s1503928080_11919
000829528 520__ $$aMotivation.The central nervous system is subdivided in different parts connected to each other via multiple paths. The directional neuronal outgrowth is fundamental for the functionality of this complex organ and influenced by various environmental conditions like signaling molecules and chemical gradients. The study of the emerging functionality of the brain is impeded by the complex structure and the manifold influencing factors. Formation of neural network in vitro is one way to decipher the performance of hierarchically organized neuronal networks. In this study, we developed a simple and effective platform to pattern neurons in neural circuitries by focussing on the influence of the geometrical structure on these network formations. By reducing the number of interconnections, the impact of the neuronal orientation can be examined in detail. Our idea was, that with a controlled geometry we can control the anatomy of the network and thereby the resulting signal propagation directionality.Material and Methods.We created interconnected networks of a few hundred neurons utilizing the microcontact printing technique. In this process, cell-adhesive protein patterns consisting of a mixture of poly-L-lysine (PLL) and laminin are transferred onto a hydrophobic, cell-repellent glass surface. We chose a triangular shaped pattern as single building block. Multiples of these single triangles are stringed together to form a loop. Dissociated rat embryonic neurons (E18) are grown on these modified substrates for at least 14 days in vitro and form structured networks. We investigated the influence of the geometrical parameters on the directionality of neuronal populations. Therefore neuronal cultures were analyzed with immunofluorescence stainings against MAP2 (somatodendritic marker) and the axon specific marker Anti-200 kD neurofilament Heavy. Additionally Calcium imaging with the fluorescent dye Fluo-4-AM was performed to investigate the spontaneous neuronal activity. To further analyze the electrical signal transmission among and within these populations we apply proteins patterns onto Multi-electrode Arrays (MEAs). This enables tracing single cell activity as well as network activity with up to 64 electrodes simultaneously. MEAs with electrode diameters of 24 µm were fabricated in the clean room facilities of Forschungszentrum Jülich (fig. A, B). Signal recordings were performed with a low-noise amplifier system developed in our institute.Results.The results of the immunofluorescence stainings showed a polar anatomy of the shaped networks (fig. D). Spontaneous neuronal activity analyzed with Calcium Imaging revealed that the anatomy of the networks can be used to predict the direction of the signal transmission. The analysis of connected populations on MEAs (fig. C) confirm our observation that the geometry of the given pattern has a major impact on the directionality of signal propagation.Discussion.Our results demonstrate that the shape of transferred protein patterns has a great influence on the emerging higher network structure and its direction of signal propagation. In conclusion the orientation of connected neuronal networks and their information processing can be controlled by choosing the right geometrical parameters. As an outlook we want to further analyze geometrical parameters affecting the hierarchy of the network such as the size of a single building block.Conclusion.Here, we show a simple and effective platform to pattern primary neurons in hierarchical populations for long-term study of neural circuitry. By interfacing these circuitries with MEA devices will allow to build neuron based logic devices for computational applications.
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000829528 588__ $$aDataset connected to CrossRef
000829528 7001_ $$0P:(DE-HGF)0$$aJonas, Albers$$b1
000829528 7001_ $$0P:(DE-Juel1)128713$$aOffenhäusser, Andreas$$b2
000829528 773__ $$0PERI:(DE-600)2411902-7$$a10.3389/conf.fnins.2016.93.00080$$gVol. 10$$v10$$x1662-453X$$y2016
000829528 8564_ $$uhttp://www.frontiersin.org/10.3389/conf.fnins.2016.93.00080/event_abstract?sname=MEA_Meeting_2016_|10th_International_Meeting_on_Substrate-Integrated_Electrode_Arrays
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000829528 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)128713$$aForschungszentrum Jülich$$b2$$kFZJ
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000829528 9141_ $$y2017
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