An algorithm for neurite outgrowth reconstruction

Weaver, Christina M.; Pinezich, John D.; Lindquist, W. Brent; Vazquez, Marcelo E.

doi:10.1016/s0165-0270(03)00017-7

Cited by 31 publications

(29 citation statements)

References 18 publications

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“…Measurements were made in exported eight-bit TIFF files using the image analysis software MetaMorph. Maximum neurite length was determined as follows: the MetaMorph "draw" function was used to draw a straight line from the border of the explants to the tip of the longest neurite or neurite bundle at 30 different locations covering each of the four quadrants of explants, as described previously (Weaver et al, 2003). Maximum neurite length was defined as the average of these 30 measurements, and the values were calibrated in micrometers using the MetaMorph "calibrate" function.…”

Section: Methodsmentioning

confidence: 99%

A Role for Synaptotagmin VII-Regulated Exocytosis of Lysosomes in Neurite Outgrowth from Primary Sympathetic Neurons

Arantes¹,

Andrews²

2006

J. Neurosci.

144

134

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Neurite outgrowth is mediated by the exocytosis of intracellular vesicles at the tips of elongating neuronal processes. The lysosomal vesicle-associated soluble N-ethylmaleimide-sensitive factor attachment protein receptor tetanus neurotoxin insensitive vesicleassociated membrane protein (TI-VAMP)/VAMP7 was previously implicated in membrane fusion events mediating neurite outgrowth, but the participation of lysosomes in this exocytic process has remained unclear. Here, we show that VAMP7 and the lysosomal glycoprotein Lamp1 extensively colocalize in vesicles present throughout the soma and neurite outgrowths of primary sympathetic neurons. Synaptotagmin VII (Syt VII), a Ca 2ϩ -sensing synaptotagmin isoform previously shown to interact with VAMP7 during lysosomal exocytosis in fibroblasts, was detected on a subset of these lysosomal glycoprotein 1 (Lamp1)/VAMP7-positive neuronal vesicles. Ionophore-stimulated exocytosis triggered exposure of the luminal domains of both Lamp1 and Syt VII at overlapping sites on the neuronal surface, indicating that the Syt VII-containing lysosomal compartments fuse with the plasma membrane in response to [Ca 2ϩ ] i elevation. To determine whether Syt VII was required for the exocytic events mediating neurite extension, we followed the development of superior cervical ganglion neurons explanted from Syt VII-deficient mice. The results revealed a marked defect in neurite outgrowth and arborization, suggesting that Ca 2ϩ -dependent, Syt VII-regulated exocytosis of late endosomes/lysosomes plays a role in the addition of new membrane to developing neurite extensions.

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Section: Methodsmentioning

confidence: 99%

A Role for Synaptotagmin VII-Regulated Exocytosis of Lysosomes in Neurite Outgrowth from Primary Sympathetic Neurons

Arantes¹,

Andrews²

2006

J. Neurosci.

144

134

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“…In phasecontrast microscopy too, the cell bodies often yield more contrast than their processes, in which case they can be segmented simply by means of intensity thresholding (33,34). If, on the other hand, the somas cannot be distinguished solely based on intensity, they may be discriminated by applying suitable (binary or grayscale) morphological filtering (35)(36)(37)(38).…”

Section: Soma Segmentationmentioning

confidence: 99%

Neuron tracing in perspective

2010

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The study of the structure and function of neuronal cells and networks is of crucial importance in the endeavor to understand how the brain works. A key component in this process is the extraction of neuronal morphology from microscopic imaging data. In the past four decades, many computational methods and tools have been developed for digital reconstruction of neurons from images, with limited success. As witnessed by the growing body of literature on the subject, as well as the organization of challenging competitions in the field, the quest for a robust and fully automated system of more general applicability still continues. The aim of this work, is to contribute by surveying recent developments in the field for anyone interested in taking up the challenge. Relevant aspects discussed in the article include proposed image segmentation methods, quantitative measures of neuronal morphology, currently available software tools for various related purposes, and morphology databases. ' 2010 International Society for Advancement of Cytometry

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“…Several investigators have abandoned tracing altogether and have resorted to heuristic approaches to estimate the morphometric features of interest (11)(12)(13). An alternative, vectorial-tracking technique, published just before completion of our manuscript, was successfully applied to images of retinal explants (14); however, it does not address the just mentioned problems seen in our type of images.…”

mentioning

confidence: 99%

Design and validation of a tool for neurite tracing and analysis in fluorescence microscopy images

et al. 2004

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Background: For the investigation of the molecular mechanisms involved in neurite outgrowth and differentiation, accurate and reproducible segmentation and quantification of neuronal processes are a prerequisite. To facilitate this task, we developed a semiautomatic neurite tracing technique. This article describes the design and validation of the technique. Methods:The technique was compared to fully manual delineation. Four observers repeatedly traced selected neurites in 20 fluorescence microscopy images of cells in culture, using both methods. Accuracy and reproducibility were determined by comparing the tracings to highresolution reference tracings, using two error measures. Labor intensiveness was measured in numbers of mouse clicks required. The significance of the results was determined by a Student t-test and by analysis of variance. Results: Both methods slightly underestimated the true neurite length, but the differences were not unanimously

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An algorithm for neurite outgrowth reconstruction

Cited by 31 publications

References 18 publications

A Role for Synaptotagmin VII-Regulated Exocytosis of Lysosomes in Neurite Outgrowth from Primary Sympathetic Neurons

A Role for Synaptotagmin VII-Regulated Exocytosis of Lysosomes in Neurite Outgrowth from Primary Sympathetic Neurons

Neuron tracing in perspective

Design and validation of a tool for neurite tracing and analysis in fluorescence microscopy images

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