a-simple-microfluidic-chip-for-long-term-growth-imaging

A Simple Microfluidic Chip for Long-Term Growth and Imaging of <em>Caenorhabditis elegans</em>

Caenorhabditis elegans (C. elegans) have proved to be a valuable model system for studying developmental and cell biological processes. Understanding ...

TIFR

InSTEM

Manipal Academy of Higher Education

Microtubules form important cytoskeletal structures that play a role in establishing and maintaining neuronal polarity, regulating neuronal morphology, transporting cargo, and scaffolding signaling molecules to form signaling hubs. Within a neuronal cell, microtubules are found to have variable lengths and can be both stable and dynamic. Microtubule associated proteins, post-translational modifications of tubulin subunits, microtubule severing enzymes, and signaling molecules are all known to influence both stable and dynamic pools of microtubules. Microtubule dynamics, the process of interconversion between stable and dynamic pools, and the proportions of these two pools have the potential to influence a wide variety of cellular processes. Reduced microtubule stability has been observed in several neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), Amyotrophic Lateral Sclerosis (ALS), and tauopathies like Progressive Supranuclear Palsy. Hyperstable microtubules, as seen in Hereditary Spastic Paraplegia (HSP), also lead to neurodegeneration. Therefore, the ratio of stable and dynamic microtubules is likely to be important for neuronal function and perturbation in microtubule dynamics might contribute to disease progression. 

Neurodegeneration and microtubule dynamics: death by a thousand cuts.

[This corrects the article on p. 343 in vol. 9, PMID: 26441521.]. 

Corrigendum: Neurodegeneration and microtubule dynamics: death by a thousand cuts.

The long cellular architecture of neurons requires regulation in part through transport and anchoring events to distribute intracellular organelles. During development, cellular and subcellular events such as organelle additions and their recruitment at specific sites on the growing axons occur over different time scales and often show interanimal variability thus making it difficult to identify specific phenomena in population averages. To measure the variability in subcellular events such as organelle positions, we developed a microfluidic device to feed and immobilize  for high-resolution imaging over several days. The microfluidic device enabled long-term imaging of individual animals and allowed us to investigate organelle density using mitochondria as a testbed in a growing neuronal process  Subcellular imaging of an individual neuron in multiple animals, over 36 h in our microfluidic device, shows the addition of new mitochondria along the neuronal process and an increase in the accumulation of synaptic vesicles (SVs) at synapses. Long-term imaging of individual  touch receptor neurons (TRNs) shows that the addition of new mitochondria takes place along the entire neuronal process length at a rate of ∼0.6 mitochondria/h. The threshold for the addition of a new mitochondrion occurs when the average separation between the two preexisting mitochondria exceeds 24 μm. Our assay provides a new opportunity to move beyond simple observations obtained from  assays to allow the discovery of genes that regulate positioning of mitochondria in neurons.

Tracking Mitochondrial Density and Positioning along a Growing Neuronal Process in Individual  Neuron Using a Long-Term Growth and Imaging Microfluidic Device.

Article	Year
Neurodegeneration and microtubule dynamics: death by a thousand cuts. Frontiers in cellular neuroscience\| PubMed ID: 26441521	2015
Corrigendum: Neurodegeneration and microtubule dynamics: death by a thousand cuts. Frontiers in cellular neuroscience\| PubMed ID: 26903811	2016
Tracking Mitochondrial Density and Positioning along a Growing Neuronal Process in Individual Neuron Using a Long-Term Growth and Imaging Microfluidic Device. eNeuro\| PubMed ID: 34035072	2021

Jyoti Dubey

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