Growth of<i>Caenorhabditis elegans</i>in Defined Media Is Dependent on Presence of Particulate Matter

Flavel, Matthew; Mechler, Ádám; Shahmiri, Mahdi; Mathews, Elizabeth R.; Franks, Ashley E.; Chen, Weisan; Zanker, Damien; Xian, Bo; Gao, Shan; Luo, Jing; Tegegne, Surafel M.; Doneski, Christian; Jois, Markandeya

doi:10.1534/g3.117.300325

Cited by 30 publications

(32 citation statements)

References 36 publications

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“…This was later supported by two studies in which pure precipitated proteins were used [68, 72, 73]. Likewise, C. elegans requires particulate matter to successfully take up the nutrients present in axenic medium [74]. By subjecting the medium to 0.22-μm filtration, it retains a very low concentration of small-sized particles that worms cannot ingest but expel with liquid, resulting in impediment of worm growth [74].…”

Section: Caloric Compounds and Building Blocksmentioning

confidence: 99%

The nutritional requirements of Caenorhabditis elegans

2019

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Animals require sufficient intake of a variety of nutrients to support their development, somatic maintenance and reproduction. An adequate diet provides cell building blocks, chemical energy to drive cellular processes and essential nutrients that cannot be synthesised by the animal, or at least not in the required amounts. Dietary requirements of nematodes, including Caenorhabditis elegans have been extensively studied with the major aim to develop a chemically defined axenic medium that would support their growth and reproduction. At the same time, these studies helped elucidating important aspects of nutrition-related biochemistry and metabolism as well as the establishment of C. elegans as a powerful model in studying evolutionarily conserved pathways, and the influence of the diet on health.

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Section: Caloric Compounds and Building Blocksmentioning

confidence: 99%

The nutritional requirements of Caenorhabditis elegans

2019

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“…The higher throughput methods described here should allow investigators to take even better advantage of some of the strengths of this model system; its optical transparency, its short life cycle, its full editable genome, and its eminent utility in forward and reverse genetic screens. Moreover, the recent development of synthetic food medium as an alternative to the typically used OP50 E. coli strain has should allow execution of chemical genetic screens, using for example, FDA approved pharmaceuticals packaged into liposomes for delivery into the worm body via the alimentary canal (FLAVEL et al 2018).…”

Section: Using C Elegans To Study Intron Functionmentioning

confidence: 99%

In vivomeasurements reveal a single 5’ intron is sufficient to increase protein expression level inC. elegans

Sands

Battaglia

et al. 2018

Preprint

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Introns can increase gene expression levels using a variety of mechanisms collectively referred to as Intron Mediated Enhancement (IME). To date, the magnitude of IME has been quantified in human cell culture and plant models by comparing intronless reporter gene expression levels to those of intron-bearing reporter genes in vitro (mRNA, Western Blots, protein activity), using genome editing technologies that lacked full control of locus and copy number. Here, for the first time, we quantified IME in vivo, in terms of protein expression levels, using fluorescent reporter proteins expressed from a single, defined locus in Caenorhabditis elegans. To quantify the magnitude of IME, we developed a microfluidic chip-based workflow to mount and image individual animals, including software for operation and image processing. We used this workflow to systematically test the effects of position, number and sequence of introns on two different proteins, mCherry and mEGFP, driven by two different promoters, vit-2 and hsp-90. We found the three canonical synthetic introns commonly used in C. elegans transgenes increased mCherry protein concentration by approximately 50%. The naturally-occurring introns found in hsp-90 also increased mCherry expression level by about 50%. Furthermore, and consistent with prior results examining mRNA levels, protein activity or phenotypic rescue, we found that a single, natural or synthetic, 5' intron was sufficient for the full IME effect while a 3' intron was not. IME was also affected by protein coding sequence (50% for mCherry and 80% for mEGFP) but not strongly affected by promoter 46% for hsp-90 and 54% for the stronger vit-2. Our results show that IME of protein expression in C. elegans is affected by intron position and contextual coding sequence surrounding the introns, but not greatly by promoter strength. Our combined controlled transgenesis and microfluidic screening approach should facilitate screens for factors affecting IME and other intron-dependent processes. + equal contribution

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“…However, we used live OP50 bacteria that could potentially metabolize and degrade a compound thereby lowering its availability to the animal. In future studies the use of dead bacteria as a food source could further maximize drug uptake or in alternative, axenic food could replace the bacteria now that this nutrient has been developed in granular form that the worms can eat more easily [35]. Mutant worms with altered cuticle permeability such as bus-5 (br19) also represent a valid alternative.…”

Section: Discussionmentioning

confidence: 99%

A novel assay for drug screening that utilizes the heat shock response of Caenorhabditis elegans nematodes

et al. 2020

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Biological organisms respond to environmental stressors by recruiting multiple cellular cascades that act to mitigate damage and ultimately enhance survival. This implies that compounds that interact with any of those pathways might improve organism's survival. Here, we report on an initial attempt to develop a drug screening assay based on the heat shock (HS) response of Caenorhabditis elegans nematodes. The protocol works by subjecting the worms to two HS conditions in the absence/presence of the test compounds. Post-heat shock survival is quantified manually or in semi-automatic manner by analyzing z-stack pictures. We blindly screened a cassette of 72 compounds in different developmental stages provided by Eli Lilly through their Open Innovation Drug Discovery program. The analysis indicated that, on average, therapeutically useful drugs increase survival to HS compared to compounds used in non-clinical settings. We developed a formalism that estimates the probability of a compound to enhance survival based on a comparison with a set of parameters calculated from a pool of 35 FDA-approved drugs. The method correctly identified the developmental stages of the Lilly compounds based on their relative abilities to enhance survival to the HS. Taken together these data provide proof of principle that an assay that measures the HS response of C. elegans can offer physiological and pharmacological insight in a cost-and time-efficient manner.

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Growth ofCaenorhabditis elegansin Defined Media Is Dependent on Presence of Particulate Matter

Cited by 30 publications

References 36 publications

The nutritional requirements of Caenorhabditis elegans

The nutritional requirements of Caenorhabditis elegans

In vivomeasurements reveal a single 5’ intron is sufficient to increase protein expression level inC. elegans

A novel assay for drug screening that utilizes the heat shock response of Caenorhabditis elegans nematodes

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