HCA-Vision: Automated Neurite Outgrowth Analysis

Wang, Dadong; Lagerstrom, Ryan; Sun, Changming; Bishof, Leanne; Valotton, Pascal; Götte, Marjo

doi:10.1177/1087057110382894

Cited by 44 publications

(20 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In comparison to freely available tools, commercial software solutions, such as HCA-Vision [15] can offer more detailed analysis on the basis of single cell measurements, and report detailed branch patterns in individual neurons. Such analysis can be beneficial for studying low-density cultures, in which neurite arbors are well separated between individual neurons.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2011

View full text Add to dashboard Cite

BackgroundTo date, some of the most useful and physiologically relevant neuronal cell culture systems, such as high density co-cultures of astrocytes and primary hippocampal neurons, or differentiated stem cell-derived cultures, are characterized by high cell density and partially overlapping cellular structures. Efficient analytical strategies are required to enable rapid, reliable, quantitative analysis of neuronal morphology in these valuable model systems.ResultsHere we present the development and validation of a novel bioinformatics pipeline called NeuriteQuant. This tool enables fully automated morphological analysis of large-scale image data from neuronal cultures or brain sections that display a high degree of complexity and overlap of neuronal outgrowths. It also provides an efficient web-based tool to review and evaluate the analysis process. In addition to its built-in functionality, NeuriteQuant can be readily extended based on the rich toolset offered by ImageJ and its associated community of developers. As proof of concept we performed automated screens for modulators of neuronal development in cultures of primary neurons and neuronally differentiated P19 stem cells, which demonstrated specific dose-dependent effects on neuronal morphology.ConclusionsNeuriteQuant is a freely available open-source tool for the automated analysis and effective review of large-scale high-content screens. It is especially well suited to quantify the effect of experimental manipulations on physiologically relevant neuronal cultures or brain sections that display a high degree of complexity and overlap among neurites or other cellular structures.

show abstract

Section: Resultsmentioning

confidence: 99%

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

et al. 2011

View full text Add to dashboard Cite

show abstract

“…The length of the neurites was quantified using HCA-Vision software (CSIRO, North Ryde, NSW, Australia). The neurite quantification procedure, which involved neuron body detection, neurite detection, and neurite analysis, was performed as previously described [28]. The neurite length obtained under control conditions (i.e., in the absence of NGF and PACAP) was subtracted from each treatment condition.…”

Section: Methodsmentioning

confidence: 99%

c-Jun N-terminal kinase in synergistic neurite outgrowth in PC12 cells mediated through P90RSK

et al. 2013

View full text Add to dashboard Cite

BackgroundSynergistic multi-ligand treatments that can induce neuronal differentiation offer valuable strategies to regulate and modulate neurite outgrowth. Whereas the signaling pathways mediating single ligand-induced neurite outgrowth, such as Akt, extracellular signal-regulated kinase (Erk), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (P38), have been extensively studied, the mechanisms underlying multi-ligand synergistic neurite outgrowth are poorly understood. In an attempt to gain insight into synergistic neurite outgrowth, PC12 cells were treated with one of three combinations: pituitary adenylate cyclase-activating peptide (PACAP) with epidermal growth factor (EP), basic fibroblast growth factor (FP), or nerve growth factor (NP) and then challenged with the appropriate kinase inhibitors to assess the signaling pathways involved in the process.ResultsResponse surface analyses indicated that synergistic neurite outgrowth was regulated by distinct pathways in these systems. Synergistic increases in the phosphorylation of Erk and JNK, but not Akt or P38, were observed with the three growth factor-PACAP combinations. Unexpectedly, we identified a synergistic increase in JNK phosphorylation, which was involved in neurite outgrowth in the NP and FP, but not EP, systems. Inhibition of JNK using the SP600125 inhibitor reduced phosphorylation of 90 kDa ribosomal S6 kinase (P90RSK) in the NP and FP, but not EP, systems. This suggested the involvement of P90RSK in mediating the differential effects of JNK in synergistic neurite outgrowth.ConclusionsTaken together, these findings reveal the involvement of distinct signaling pathways in regulating neurite outgrowth in response to different synergistic growth factor-PACAP treatments. Our findings demonstrate a hitherto unrecognized mechanism of JNK-P90RSK in mediating synergistic neurite outgrowth induced by the co-treatment of growth factors and PACAP.

show abstract

“…To accomplish this, high content screening systems can be employed, such as the PerkinElmer Opera automated confocal microscope. For the impaired axonal outgrowth in HSP, the measurement of neurite outgrowth parameters can be automatically quantified using existing software (Wang et al, 2010). The automated identification of axonal swellings is a little more difficult because no software is currently available to do so.…”

Section: Challenges and Future Directionsmentioning

confidence: 99%

Modeling axonal defects in hereditary spastic paraplegia with human pluripotent stem cells

Denton

Shah³

et al. 2016

Front. Biol.

View full text Add to dashboard Cite

BACKGROUND Cortical motor neurons, also known as upper motor neurons, are large projection neurons whose axons convey signals to lower motor neurons to control the muscle movements. Degeneration of cortical motor neuron axons is implicated in several debilitating disorders, including hereditary spastic paraplegia (HSP) and amyotrophic lateral sclerosis (ALS). Since the discovery of the first HSP gene, SPAST that encodes spastin, over 70 distinct genetic loci associated with HSP have been identified. How the mutations of these functionally diverse genes result in axonal degeneration and why certain axons are affected in HSP remains largely unknown. The development of induced pluripotent stem cell (iPSC) technology has provided researchers an excellent resource to generate patient-specific human neurons to model human neuropathologic processes including axonal defects. METHODS In this article, we will frst review the pathology and pathways affected in the common forms of HSP subtypes by searching the PubMed database. We will then summurize the findings and insights gained from studies using iPSC-based models, and discuss the challenges and future directions. RESULTS HSPs, a heterogeneous group of genetic neurodegenerative disorders, are characterized by lower extremity weakness and spasticity that result from retrograde axonal degeneration of cortical motor neurons. Recently, iPSCs have been generated from several common forms of HSP including SPG4, SPG3A, and SPG11 patients. Neurons derived from HSP iPSCs exhibit disease-relevant axonal defects, such as impaired neurite outgrowth, increased axonal swellings, and reduced axonal transport. CONCLUSION These patient-derived neurons offer unique tools to study the pathogenic mechanisms and explore the treatments for rescuing axonal defects in HSP, as well as other diseases involving axonopathy.

show abstract

HCA-Vision: Automated Neurite Outgrowth Analysis

Cited by 44 publications

References 11 publications

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

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

c-Jun N-terminal kinase in synergistic neurite outgrowth in PC12 cells mediated through P90RSK

Modeling axonal defects in hereditary spastic paraplegia with human pluripotent stem cells

Contact Info

Product

Resources

About