CO<sub>2</sub>and compressive immobilization of C. elegans on-chip

Chokshi, Trushal Vijaykumar; Ben‐Yakar, Adela; Chronis, Nikos

doi:10.1039/b807345g

Cited by 126 publications

(135 citation statements)

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“…Immobilization has been achieved mechanically, thermally, or chemically using pressure (Hulme et al, 2007(Hulme et al, , 2010, gelation (Krajniak and Lu, 2010), low temperatures (Chung et al, 2008), or carbondioxide (Chokshi et al, 2009).…”

Section: An Improved System For Imaging C Elegans Larvaementioning

confidence: 99%

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Bringmann

2011

Journal of Neuroscience Methods

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a b s t r a c tCaenorhabditis elegans larvae display specific behavior and development that is not observed in adults. For example, larvae go through a molting cycle that includes a sleep-like state prior to the molt. The study of these processes requires high-resolution long-term observation of individual animals. Here we describe a method for simultaneous culture and observation of several individual young C. elegans larvae inside agarose hydrogel-based arrayed microcompartments. We used agarose hydrogel microcompartments to observe and quantify larval specific sleep-wake-like behavior and to observe neuronal rewiring using confocal fluorescence microscopy without acute immobilization. We found no behavioral aberrations caused by area restriction. We show that worms cultured inside hydrogel microcompartments develop into normal adults. Thus, hydrogel microcompartments appear useful for long-term observation of larval behavior and development.

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Section: An Improved System For Imaging C Elegans Larvaementioning

confidence: 99%

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Bringmann

2011

Journal of Neuroscience Methods

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“…In addition, anesthetics need several minutes to take effect, and recovery of nematodes from anesthesia requires exchange of media without losing animals, all of which hinder high-throughput screening. Other techniques that can be used to reversibly immobilize C. elegans include trapping of nematodes in wedge-shaped microchannels (13), cooling (14,15), and exposure to CO 2 (16,17). However, the physiological effects of exposure to low temperatures and CO 2 remain uncharacterized for many biological processes.…”

Section: Elegans | Chemical Screen | Microfluidicsmentioning

confidence: 99%

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

Samara

Rohde

Gilleland

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

110

114

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Discovery of molecular mechanisms and chemical compounds that enhance neuronal regeneration can lead to development of therapeutics to combat nervous system injuries and neurodegenerative diseases. By combining high-throughput microfluidics and femtosecond laser microsurgery, we demonstrate for the first time largescale in vivo screens for identification of compounds that affect neurite regeneration. We performed thousands of microsurgeries at single-axon precision in the nematode Caenorhabditis elegans at a rate of 20 seconds per animal. Following surgeries, we exposed the animals to a hand-curated library of approximately one hundred small molecules and identified chemicals that significantly alter neurite regeneration. In particular, we found that the PKC kinase inhibitor staurosporine strongly modulates regeneration in a concentration-and neuronal type-specific manner. Two structurally unrelated PKC inhibitors produce similar effects. We further show that regeneration is significantly enhanced by the PKC activator prostratin. C. elegans | chemical screen | microfluidicsT he ability of neurons in the adult mammalian central nervous system to regenerate their axons after injury is extremely limited, which has been attributed to both extrinsic signals of the inhibitory glial environment (1) as well as intrinsic neuronal factors (2-4). The discovery of cell-permeable small molecules that modulate axon regrowth can potentiate the development of efficient therapeutic treatments for spinal cord injuries, brain trauma, stroke, and neurodegenerative diseases. Identification of such molecules can also provide valuable tools for fundamental investigations of the mechanisms involved in the regeneration process. Currently, small-molecule screens for neuronal regeneration are performed in simple in vitro cell culture systems. Such screens have already revealed large numbers of chemicals that enhance regeneration and/or affect cellular morphogenesis, yet many of these hits still remain untested in vivo. In addition, most in vitro studies do not translate to animal models and also fail to reveal off-target, toxic, or lethal effects. Thus, a thorough investigation of neuronal regeneration mechanisms requires in vivo neuronal injury models.In vivo investigation of neuronal regeneration has been performed mainly in mice and rats. However, their long developmental periods, complicated genetics and biology, and expensive maintenance prevent large-scale studies on these animals. The nematode Caenorhabditis elegans is a simple, well-studied, invertebrate model-organism with a fully mapped neuronal network comprising 302 neurons. Its short developmental cycle, simple and low-cost laboratory maintenance, and genetic amenability make it an ideal model for large-scale screens, rapid identification of the molecular targets of screened compounds, and discovery of novel signaling pathways implicated in regeneration.Until recently however, the small size of C. elegans (∼50 μm in diameter) prevented its use for investigation of neuronal re...

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“…Microfluidic technology has been leveraged for various applications in nematode biology-from immobilization for imaging to laser ablation in neuronal and ageing studies [22][23][24][25]. Worm growth has also been studied using an array of chambers in which L4 worms were individually loaded to monitor development to adulthood [26], or using a device developed to observe collections of 30-40 adults worms [27].…”

Section: Introductionmentioning

confidence: 99%

A size threshold governsCaenorhabditis elegansdevelopmental progression

Uppaluri

Brangwynne

2015

Proc. R. Soc. B.

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The growth of organisms from humans to bacteria is affected by environmental conditions. However, mechanisms governing growth and size control are not well understood, particularly in the context of changes in food availability in developing multicellular organisms. Here, we use a novel microfluidic platform to study the impact of diet on the growth and development of the nematode Caenorhabditis elegans . This device allows us to observe individual worms throughout larval development, quantify their growth as well as pinpoint the moulting transitions marking successive developmental stages. Under conditions of low food availability, worms grow very slowly, but do not moult until they have achieved a threshold size. The time spent in larval stages can be extended by over an order of magnitude, in agreement with a simple threshold size model. Thus, a critical worm size appears to trigger developmental progression, and may contribute to prolonged lifespan under dietary restriction.

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CO₂and compressive immobilization of C. elegans on-chip

Cited by 126 publications

References 35 publications

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

A size threshold governsCaenorhabditis elegansdevelopmental progression

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CO2and compressive immobilization of C. elegans on-chip

Cited by 126 publications

References 35 publications

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Agarose hydrogel microcompartments for imaging sleep- and wake-like behavior and nervous system development in Caenorhabditis elegans larvae

Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration

A size threshold governsCaenorhabditis elegansdevelopmental progression

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CO₂and compressive immobilization of C. elegans on-chip