Selective pharmacological manipulation of cortical–thalamic co-cultures in a dual-compartment device

Kanagasabapathi, Thirukumaran T.; Franco, María Clara; Barone, Rocco Andrea; Martinoia, Sergio; Wadman, Wytse J.; Decré, M.

doi:10.1016/j.jneumeth.2012.12.019

Cited by 30 publications

(27 citation statements)

References 35 publications

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“…Axon-selective microtunnels of 10 μm width were constructed to visualize bundles of axons [3, 14] and for analysis of CNS axonal injury and regeneration [15]. Other work with two different brain regions of thalamus and cortex followed later [16], but did not monitor axon activity with electrodes in the tunnels as in our work with pairs of DG-CA3 and CA3-CA1 [8]. Electrical activity of the axons was recorded from a single electrode [17] and multisites [4, 8, 18] through the tunnels with higher signal-to-noise ratio value [5] than conventional open-access substrate-embedded microelectrodes.…”

Section: Discussionmentioning

confidence: 99%

Narrow microtunnel technology for the isolation and precise identification of axonal communication among distinct hippocampal subregion networks

Narula¹,

Ruíz²,

McQuaide³

et al. 2017

PLoS ONE

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Communication between different sub regions of the hippocampus is fundamental to learning and memory. However accurate knowledge about information transfer between sub regions from access to the activity in individual axons is lacking. MEMS devices with microtunnels connecting two sub networks have begun to approach this problem but the commonly used 10 μm wide tunnels frequently measure signals from multiple axons. To reduce this complexity, we compared polydimethylsiloxane (PDMS) microtunnel devices each with a separate tunnel width of 2.5, 5 or 10 μm bridging two wells aligned over a multi electrode array (MEA). Primary rat neurons were grown in the chambers with neurons from the dentate gyrus on one side and hippocampal CA3 on the other. After 2–3 weeks of culture, spontaneous activity in the axons inside the tunnels was recorded. We report electrophysiological, exploratory data analysis for feature clustering and visual evidence to support the expectation that 2.5 μm wide tunnels have fewer axons per tunnel and therefore more clearly delineated signals than 10 or 5 μm wide tunnels. Several measures indicated that fewer axons per electrode enabled more accurate detection of spikes. A clustering analysis comparing the variations of spike height and width for different tunnel widths revealed tighter clusters representing unique spikes with less height and width variation when measured in narrow tunnels. Wider tunnels tended toward more diffuse clusters from a continuum of spike heights and widths. Standard deviations for multiple cluster measures, such as Average Dissimilarity, Silhouette Value (S) and Separation Factor (average dissimilarity/S value), support a conclusion that 2.5 μm wide tunnels containing fewer axons enable more precise determination of individual action potential peaks, their propagation direction, timing, and information transfer between sub networks.

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Section: Discussionmentioning

confidence: 99%

Narrow microtunnel technology for the isolation and precise identification of axonal communication among distinct hippocampal subregion networks

Narula¹,

Ruíz²,

McQuaide³

et al. 2017

PLoS ONE

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“…[8][9][10][11][12][13] Microfluidic systems and procedures, that were originally developed to compartmentalise neurons to study their axons in isolation and their response to injury, 9 have led to a range of analytical applications addressing neuron/glia co-cultures, cortical/thalamic neuronal co-cultures and neuronneuron cultures, as well as studying synapse formation. 10,13,14 These systems allow for increasingly complex but ''ordered'' models of the CNS to be created in vitro, facilitating the application of localised chemical stimuli, increasing the throughput of monitored responses and reducing the amount of drugs required per experiment, a factor particularly important for screening novel or expensive compounds. This makes microfluidic systems a versatile, low-cost and useful tool to screen new drugs and/or examine the genetic changes associated with CNS conditions.…”

Section: Introductionmentioning

confidence: 99%

Chemically induced synaptic activity between mixed primary hippocampal co-cultures in a microfluidic system

Robertson

Bushell

Zagnoni

2014

Integrative Biology

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Primary neuronal cultures are an invaluable in vitro tool for examining the fundamental physiological changes that occur in diseases of the central nervous system. In this work, we have used a microfluidic device to grow twin cultures of primary hippocampal neuronal/glia cells which are synaptically connected but environmentally isolated. Immunocytochemical staining, for β-III-Tubulin and synaptophysin, indicated that the two neuronal populations were physically connected and that synapses were present. By dispensing predefined volumes of fluids into the device inlets, one culture was chemically stimulated and the consequent increase in neuronal activity in the opposing culture was monitored using calcium imaging. To optimise the experimental procedures, we validated a numerical model that estimates the concentration distribution of substances under dynamic fluidic conditions, proposing that no cross contamination of chemical stimuli occurred during the experiments. Calcium imaging and local chemical stimulation were used to confirm synaptic connectivity between the cultures. Chemical stimulation of one population, using KCl or glutamate, resulted in a significant increase of calcium events in both neurons and astrocytes of the connected population. The integration of the system and techniques described here presents a novel methodology for probing the functional synaptic connectivity between mixed primary hippocampal co-cultures, creating an in vitro testing platform for the high-throughput investigation of synaptic activity modulation either by novel compounds or in in vitro disease models.

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“…For example, Kanagasabapathi et al have created MEA-coupled compartmentalized NSCs for monitoring cortical and thalamic cell connectivity. 53,54 Various studies have also used MEA-coupled microfluidic NSCs to study the effect of biochemical cues on neural networks. 44,51,52 Smith et al have created a cantilever-based NSC that measures muscle contraction following the stimulation of a motoneuron.…”

Section: Emerging Areas and Future Directionsmentioning

confidence: 99%

Microphysiological Human Brain and Neural Systems-on-a-Chip: Potential Alternatives to Small Animal Models and Emerging Platforms for Drug Discovery and Personalized Medicine

Haring

Sontheimer

Johnson

2017

Stem Cell Rev and Rep

105

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Translational challenges associated with reductionist modeling approaches, as well as ethical concerns and economic implications associated with small animal testing, drive the need for developing microphysiological neural systems for modeling human neurological diseases, disorders and injuries. Here, we provide a comprehensive review of microphysiological neural systems on a chip (NSCs) for modeling higher order trajectories in the human nervous system. Societal, economic, and national security impacts of neurological diseases, disorders and injuries are highlighted to identify critical NSC application spaces. Hierarchical design and manufacturing of NSCs are discussed with distinction of surface- and bulk-based systems. Three broad NSC classes are identified and reviewed: microfluidic NSCs, compartmentalized NSCs, and hydrogel NSCs. Emerging areas and future directions are highlighted, including the application of 3D printing to design and manufacturing of next-generation NSCs, the use of stem cells for constructing patient-specific NSCs, and the application of human NSCs to ‘personalized neurology’. Technical hurdles and remaining challenges are discussed. This review identifies the state-of-the-art design methodologies, manufacturing approaches, and performance capabilities of NSCs. This work suggests NSCs appear poised to revolutionize the modeling of human neurological diseases, disorders and injuries.

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Selective pharmacological manipulation of cortical–thalamic co-cultures in a dual-compartment device

Cited by 30 publications

References 35 publications

Narrow microtunnel technology for the isolation and precise identification of axonal communication among distinct hippocampal subregion networks

Narrow microtunnel technology for the isolation and precise identification of axonal communication among distinct hippocampal subregion networks

Chemically induced synaptic activity between mixed primary hippocampal co-cultures in a microfluidic system

Microphysiological Human Brain and Neural Systems-on-a-Chip: Potential Alternatives to Small Animal Models and Emerging Platforms for Drug Discovery and Personalized Medicine

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