Methods and Applications of Campenot Trichamber Neuronal  Cultures for the Study of Neuroinvasive Viruses

Tierney, Wesley M.; Vicino, Ian A.; Sun, Shiqi; Chiu, Wah; Engel, Esteban A.; Taylor, Matthew P.; Hogue, Ian B.

doi:10.20944/preprints202105.0098.v1

Cited by 3 publications

(1 citation statement)

References 40 publications

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“…Uniformly placed scratches on the coated culture plate ensure that the regenerating neurites grow linearly on the coated areas in-between scratches, with minimal bending ( Figure 2 ). Notably, the rake for applying scratches is made by cementing 20 insect pins placed 200 μm apart, which yields a set of 20 parallel scratches on coated cell culture plates [ 45 , 46 , 47 ].…”

Section: Transection Injury Modelsmentioning

confidence: 99%

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

Varier

Raju

Madhusudanan

et al. 2022

IJMS

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Nerve axonal injury and associated cellular mechanisms leading to peripheral nerve damage are important topics of research necessary for reducing disability and enhancing quality of life. Model systems that mimic the biological changes that occur during human nerve injury are crucial for the identification of cellular responses, screening of novel therapeutic molecules, and design of neural regeneration strategies. In addition to in vivo and mathematical models, in vitro axonal injury models provide a simple, robust, and reductionist platform to partially understand nerve injury pathogenesis and regeneration. In recent years, there have been several advances related to in vitro techniques that focus on the utilization of custom-fabricated cell culture chambers, microfluidic chamber systems, and injury techniques such as laser ablation and axonal stretching. These developments seem to reflect a gradual and natural progression towards understanding molecular and signaling events at an individual axon and neuronal-soma level. In this review, we attempt to categorize and discuss various in vitro models of injury relevant to the peripheral nervous system and highlight their strengths, weaknesses, and opportunities. Such models will help to recreate the post-injury microenvironment and aid in the development of therapeutic strategies that can accelerate nerve repair.

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Section: Transection Injury Modelsmentioning

confidence: 99%

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

Varier

Raju

Madhusudanan

et al. 2022

IJMS

View full text Add to dashboard Cite

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High-Resolution Imaging of Mitochondria and Mitochondrial Nucleoids in Differentiated SH-SY5Y Cells

Annuario

Vagnoni

2022

Methods in Molecular Biology

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Mitochondria are highly dynamic organelles which form intricate networks with complex dynamics. Mitochondrial transport and distribution are essential to ensure proper cell function, especially in cells with an extremely polarised morphology such as neurons. A layer of complexity is added when considering mitochondria have their own genome, packaged into nucleoids. Major mitochondrial morphological transitions, for example mitochondrial division, often occur in conjunction with mitochondrial DNA (mtDNA) replication and changes in the dynamic behaviour of the nucleoids. However, the relationship between mtDNA dynamics and mitochondrial motility in the processes of neurons has been largely overlooked. In this chapter, we describe a method for live imaging of mitochondria and nucleoids in differentiated SH-SY5Y cells by instant structured illumination microscopy (iSIM). We also include a detailed protocol for the differentiation of SH-SY5Y cells into cells with a pronounced neuronal-like morphology and show examples of coordinated mitochondrial and nucleoid motility in the long processes of these cells.

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Detailed Imaging of Mitochondrial Transport and Precise Manipulation of Mitochondrial Function with Genetically Encoded Photosensitizers in Adult Drosophila Neurons

Mattedi

Chennell

Vagnoni

2022

Methods in Molecular Biology

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Precise distribution of mitochondria is essential for maintaining neuronal homeostasis. Although detailed mechanisms governing the transport of mitochondria have emerged, it is still poorly understood how the regulation of transport is coordinated in space and time within the physiological context of an organism. How alteration in mitochondrial functionality may trigger changes in organellar dynamics also remains unclear in this context. Therefore, the use of genetically encoded tools to perturb mitochondrial functionality in real time would be desirable. Here we describe methods to interfere with mitochondrial function with high spatiotemporal precision with the use of photosensitizers in vivo in the intact wing nerve of adult Drosophila. We also provide details on how to visualize the transport of mitochondria and to improve the quality of the imaging to attain super-resolution in this tissue.

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Methods and Applications of Campenot Trichamber Neuronal Cultures for the Study of Neuroinvasive Viruses

Cited by 3 publications

References 40 publications

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

High-Resolution Imaging of Mitochondria and Mitochondrial Nucleoids in Differentiated SH-SY5Y Cells

Detailed Imaging of Mitochondrial Transport and Precise Manipulation of Mitochondrial Function with Genetically Encoded Photosensitizers in Adult Drosophila Neurons

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