NeuriteQuant: An open source toolkit for high content screens of neuronal Morphogenesis

Dehmelt, Leif; Poplawski, Gunnar; Hwang, Eric; Halpain, Shelley

doi:10.1186/1471-2202-12-100

Cited by 42 publications

(40 citation statements)

References 22 publications

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“…Immunostaining images or time lapse images were obtained with a fluorescence microscope (Nikon ECLIPSE Ti-S) interfaced with a digital charge-coupled device camera and an image analysis system or confocal microscope analysis using Carl Zeiss Laser Scanning Confocal Microscope Pascal 5 LSM and Pascal 5 LSM software Version 3.2. Dendrite elongation was assessed following MAP2 staining followed by NeuriteQuant analysis 37 .…”

Section: Methodsmentioning

confidence: 99%

Transfer of mitochondria from astrocytes to neurons after stroke

Hayakawa

Esposito

Wang

et al. 2016

Nature

1,046

915

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Recently, it was suggested that neurons can release and transfer damaged mitochondria to astrocytes for disposal and recycling 1. This ability to exchange mitochondria may represent a potential mode of cell-cell signaling in the central nervous system (CNS). Here, we show that astrocytes can also release functional mitochondria that enter into neurons. Astrocytic release of extracellular mitochondria particles was mediated by a calcium-dependent mechanism involving CD38/cyclic ADP ribose signaling. Transient focal cerebral ischemia in mice induced astrocytic mitochondria entry to adjacent neurons that amplified cell survival signals. Suppression of CD38 signaling with siRNA reduced extracellular mitochondria transfer and worsened neurological outcomes. These findings suggest a new mitochondrial mechanism of neuroglial crosstalk that may contribute to endogenous neuroprotective and neurorecovery mechanisms after stroke.

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Section: Methodsmentioning

confidence: 99%

Transfer of mitochondria from astrocytes to neurons after stroke

Hayakawa

Esposito

Wang

et al. 2016

Nature

1,046

915

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“…50–55 These techniques can be used to acquire simple morphometric measures, such as average neurite length, which were not evaluated in this study due to the complex neurite network formed by the dense neuronal cultures. However, our technique can be used to approximate the density of complex neuronal networks based on the percent area that the neurites cover in a specific region of interest.…”

Section: Discussionmentioning

confidence: 99%

Morphometric assessment of toxicant induced neuronal degeneration in full and restricted contact co-cultures of embryonic cortical rat neurons and astrocytes: Using m-Dinitrobezene as a model neurotoxicant

Dixon

Philbert

2015

Toxicology in Vitro

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With m-Dinitrobenzene (m-DNB) as a selected model neurotoxicant, we demonstrate how to assess neurotoxicity, using morphology based measurement of neurite degeneration, in a conventional “full-contact” and a modern “restricted-contact” co-culture of rat cortical neurons and astrocytes. In the “full-contact” co-culture, neurons and astrocytes in complete physical contact are “globally” exposed to m-DNB. A newly emergent “restricted-contact” co-culture is attained with a microfluidic device that polarizes neuron somas and neurites into separate compartments, and the neurite compartment is “selectively” exposed to m-DNB. Morphometric analysis of the neuronal area revealed that m-DNB exposure produced no significant change in mean neuronal cell area in “full-contact” co-cultures, whereas a significant decrease was observed for neuron monocultures. Neurite elaboration into a neurite exclusive compartment in a compartmentalized microfluidic device, for both monocultures (no astrocytes) and “restricted” co-cultures (astrocytes touching neurites), decreased with exposure to increasing concentrations of m-DNB, but the average neurite area was higher in co-cultures. By using co-culture systems that more closely approach biological and architectural complexities, and the directionality of exposure found in the brain, this study provides a methodological foundation for unraveling the role of physical contact between astrocytes and neurons in mitigating the toxic effects of chemicals such as m-DNB.

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“…Cellular coverage was measured at 7 DIV by setting a threshold in ImageJ highlighting the total fluorescent cell area and quantified as a percentage of the total area of substrate; this measurement was further subdivided into neurite and soma coverage by limiting the mask threshold. Neurite length measurements were performed utilizing the NeuriteQuant toolkit in ImageJ [48]. Reported data are cell density (cells per mm 2 ) and total neurite length per area at 7 DIV.…”

Section: Methodsmentioning

confidence: 99%

Scale of Carbon Nanomaterials Affects Neural Outgrowth and Adhesion

Franca

Jao

Fang

et al. 2016

IEEE Trans.on Nanobioscience

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Carbon nanomaterials have become increasingly popular microelectrode materials for neuroscience applications. Here we study how the scale of carbon nanotubes and carbon nanofibers affect neural viability, outgrowth, and adhesion. Carbon nanotubes were deposited on glass coverslips via a layer-by-layer method with polyethylenimine (PEI). Carbonized nanofibers were fabricated by electrospinning SU-8 and pyrolyzing the nanofiber depositions. Additional substrates tested were carbonized and SU-8 thin films and SU-8 nanofibers. Surfaces were O2-plasma treated, coated with varying concentrations of PEI, seeded with E18 rat cortical cells, and examined at 3, 4, and 7 days in vitro (DIV). Neural adhesion was examined at 4 DIV utilizing a parallel plate flow chamber. At 3 DIV, neural viability was lower on the nanofiber and thin film depositions treated with higher PEI concentrations which corresponded with significantly higher zeta potentials (surface charge); this significance was drastically higher on the nanofibers suggesting that the nanostructure may collect more PEI molecules, causing increased toxicity. At 7 DIV, significantly higher neurite outgrowth was observed on SU-8 nanofiber substrates with nanofibers a significant fraction of a neuron’s size. No differences were detected for carbonized nanofibers or carbon nanotubes. Both carbonized and SU-8 nanofibers had significantly higher cellular adhesion post-flow in comparison to controls whereas the carbon nanotubes were statistically similar to control substrates. These data suggest a neural cell preference for larger-scale nanomaterials with specific surface treatments. These characteristics could be taken advantage of in the future design and fabrication of neural microelectrodes.

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NeuriteQuant: An open source toolkit for high content screens of neuronal Morphogenesis

Cited by 42 publications

References 22 publications

Transfer of mitochondria from astrocytes to neurons after stroke

Transfer of mitochondria from astrocytes to neurons after stroke

Morphometric assessment of toxicant induced neuronal degeneration in full and restricted contact co-cultures of embryonic cortical rat neurons and astrocytes: Using m-Dinitrobezene as a model neurotoxicant

Scale of Carbon Nanomaterials Affects Neural Outgrowth and Adhesion

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