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https://www.readbyqxmd.com/read/29343752/counting-caenorhabditis-elegans-protocol-optimization-and-applications-for-population-growth-and-toxicity-studies-in-liquid-medium
#1
Leona D Scanlan, Steven P Lund, Sanem Hosbas Coskun, Shannon K Hanna, Monique E Johnson, Christopher M Sims, Karina Brignoni, Patricia Lapasset, Elijah J Petersen, John T Elliott, Bryant C Nelson
The nematode Caenorhabditis elegans is used extensively in molecular, toxicological and genetics research. However, standardized methods for counting nematodes in liquid culture do not exist despite the wide use of nematodes and need for accurate measurements. Herein, we provide a simple and affordable counting protocol developed to maximize count accuracy and minimize variability in liquid nematode culture. Sources of variability in the counting process were identified and tested in 14 separate experiments...
January 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29343694/sertraline-paroxetine-and-chlorpromazine-are-rapidly-acting-anthelmintic-drugs-capable-of-clinical-repurposing
#2
Janis C Weeks, William M Roberts, Caitlyn Leasure, Brian M Suzuki, Kristin J Robinson, Heather Currey, Phurpa Wangchuk, Ramon M Eichenberger, Aleen D Saxton, Thomas D Bird, Brian C Kraemer, Alex Loukas, John M Hawdon, Conor R Caffrey, Nicole F Liachko
Parasitic helminths infect over 1 billion people worldwide, while current treatments rely on a limited arsenal of drugs. To expedite drug discovery, we screened a small-molecule library of compounds with histories of use in human clinical trials for anthelmintic activity against the soil nematode Caenorhabditis elegans. From this screen, we found that the neuromodulatory drugs sertraline, paroxetine, and chlorpromazine kill C. elegans at multiple life stages including embryos, developing larvae and gravid adults...
January 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29334121/the-multipotency-to-commitment-transition-mct-in-c-elegans-implications-for-reprogramming-from-cells-to-organs
#3
REVIEW
Erik A Spickard, Pradeep M Joshi, Joel H Rothman
In animal embryos, cells transition from a multipotential state, with the capacity to adopt multiple fates, into an irreversible, committed state of differentiation. This multipotency-to-commitment transition (MCT) is evident from experiments in which cell fate is reprogrammed by transcription factors for cell-type-specific differentiation, as has been observed extensively in C. elegans. Although factors that direct differentiation into each of the three germ layer types cannot generally reprogram cells after the MCT in this animal, transcription factors for endoderm development are able to do so in multiple differentiated cell types...
January 15, 2018: FEBS Letters
https://www.readbyqxmd.com/read/29242146/control-of-oocyte-meiotic-maturation-in-c-elegans
#4
REVIEW
Gabriela Huelgas-Morales
In virtually all sexually reproducing animals, oocytes arrest in meiotic prophase and resume meiosis in a conserved biological process called meiotic maturation. Meiotic arrest enables oocytes, which are amongst the largest cells in an organism, to grow and accumulate the necessary cellular constituents required to support embryonic development. Oocyte arrest can be maintained for a prolonged period, up to 50 years in humans, and defects in the meiotic maturation process interfere with the faithful segregation of meiotic chromosomes, representing the leading cause of human birth defects and female infertility...
December 11, 2017: Seminars in Cell & Developmental Biology
https://www.readbyqxmd.com/read/29223862/wave-regulates-cadherin-junction-assembly-and-turnover-during-epithelial-polarization
#5
Shashikala Sasidharan, Sofya Borinskaya, Falshruti Patel, Yelena Bernadskaya, Sailaja Mandalapu, Maria Agapito, Martha Soto
Actin is an integral component of epithelial apical junctions, yet the interactions of branched actin regulators with apical junction components are still not clear. Biochemical data have shown that α-catenin inhibits Arp2/3-dependent branched actin. These results suggested that branched actin is only needed at earliest stages of apical junction development. We use live imaging in developing C. elegans embryos to test models for how WAVE-induced branched actin collaborates with other apical junction proteins during the essential process of junction formation and maturation...
December 6, 2017: Developmental Biology
https://www.readbyqxmd.com/read/29217120/hydric-environmental-effects-on-turtle-development-and-sex-ratio
#6
Itzel Sifuentes-Romero, Boris M Tezak, Sarah L Milton, Jeanette Wyneken
Experimental and field studies of different turtle species suggest that moisture influences embryonic development and sex ratios, wetter substrates tend to produce more males, and drier substrates produce more females. In this study, we used Trachemys scripta elegans to test the effect of moisture on embryonic development and sex ratios. T. s. elegans eggs were incubated under different temperature and moisture regimes. We monitored embryonic development until stage 22 (after sex determination) and, for the first time, we estimated sex ratios using a male-specific transcriptional molecular marker, Sox9...
November 28, 2017: Zoology: Analysis of Complex Systems, ZACS
https://www.readbyqxmd.com/read/29192061/nude-l-regulates-dynein-at-kinetochores-but-is-dispensable-for-other-dynein-functions-in-the-c-elegans-early-embryo
#7
Patrícia A Simões, Ricardo Celestino, Ana X Carvalho, Reto Gassmann
In mitosis, the molecular motor dynein is recruited to kinetochores by the Rod-Zw10-Zwilch complex (RZZ) and Spindly to control spindle assembly checkpoint (SAC) signaling and microtubule attachment. How the ubiquitous dynein co-factors Lis1 and NudE/L contribute to these functions remains poorly understood. Here, we show that the C. elegans NudE/L homolog NUD-2 is dispensable for dynein- and LIS-1-dependent mitotic spindle assembly in the zygote. This facilitates functional characterization of kinetochore-localized NUD-2, which is recruited by the CENP-F-like proteins HCP-1/2 independently of RZZ-Spindly and dynein-LIS-1...
November 30, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29158261/the-biotin-ligating-protein-bpl-1-is-critical-for-lipid-biosynthesis-and-polarization-of-the-caenorhabditis-elegans-embryo
#8
Jason S Watts, Diane G Morton, Kenneth J Kemphues, Jennifer L Watts
Biotin is an essential cofactor for multiple metabolic reactions catalyzed by carboxylases. Biotin is covalently linked to apoproteins by holocarboxylase synthetase (HCS). Accordingly, some mutations in HCS cause holocarboxylase deficiency, a rare metabolic disorder that can be life threatening if left untreated. However, the long-term effects of HCS deficiency are poorly understood. Here, we report our investigations of bpl-1, which encodes the C. elegans ortholog of HCS. We found that mutations in the biotin-binding region of bpl-1 are maternal-effect lethal, and cause defects in embryonic polarity establishment, meiosis, and the integrity of the eggshell permeability barrier...
November 20, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29129923/ist1-regulates-escrt-iii-assembly-and-function-during-multivesicular-endosome-biogenesis-in-caenorhabditis-elegans-embryos
#9
E B Frankel, Raakhee Shankar, James J Moresco, John R Yates, Niels Volkmann, Anjon Audhya
Degradation of most integral membrane proteins is directed by the endosomal sorting complex required for transport (ESCRT) machinery, which selectively targets ubiquitin-modified cargoes into intralumenal vesicles (ILVs) within multivesicular endosomes (MVEs). To better understand the mechanisms underlying ESCRT-mediated formation of ILVs, we exploited the rapid, de novo biogenesis of MVEs during the oocyte-to-embryo transition in C. elegans. In contrast to previous models suggesting that ILVs form individually, we demonstrate that they remain tethered to one another subsequent to internalization, arguing that they bud continuously from stable subdomains...
November 13, 2017: Nature Communications
https://www.readbyqxmd.com/read/29091720/loss-of-the-caenorhabditis-elegans-pocket-protein-lin-35-reveals-muvb-s-innate-function-as-the-repressor-of-dream-target-genes
#10
Paul D Goetsch, Jacob M Garrigues, Susan Strome
The DREAM (Dp/Retinoblastoma(Rb)-like/E2F/MuvB) transcriptional repressor complex acts as a gatekeeper of the mammalian cell cycle by establishing and maintaining cellular quiescence. How DREAM's three functional components, the E2F-DP heterodimer, the Rb-like pocket protein, and the MuvB subcomplex, form and function at target gene promoters remains unknown. The current model invokes that the pocket protein links E2F-DP and MuvB and is essential for gene repression. We tested this model by assessing how the conserved yet less redundant DREAM system in Caenorhabditis elegans is affected by absence of the sole C...
November 2017: PLoS Genetics
https://www.readbyqxmd.com/read/29076351/activity-of-gallic-acid-and-its-ester-derivatives-in-caenorhabditis-elegans-and-zebrafish-danio-rerio-models
#11
Junya de Lacorte Singulani, Liliana Scorzoni, Paulo César Gomes, Ana Carolina Nazaré, Carlos Roberto Polaquini, Luis Octávio Regasini, Ana Marisa Fusco-Almeida, Maria José Soares Mendes-Giannini
AIM: Gallic acid and its ester derivatives have shown antifungal activity in vitro. This study was performed to investigate their activity against Candida albicans and their toxicity in the animal models Caenorhabditis elegans and zebrafish embryos. RESULTS: The compounds protected worms from C. albicans infection. The dodecyl gallate was the most effective. In zebrafish embryo, gallic acid and dodecyl gallate were the least toxic. CONCLUSION: Gallic acid and its ester derivatives have potential for in vivo use against C...
October 2017: Future Medicinal Chemistry
https://www.readbyqxmd.com/read/29075433/differences-in-the-genetic-control-of-early-egg-development-and-reproduction-between-c-elegans-and-its-parthenogenetic-relative-d-coronatus
#12
Christopher Kraus, Philipp H Schiffer, Hiroshi Kagoshima, Hideaki Hiraki, Theresa Vogt, Michael Kroiher, Yuji Kohara, Einhard Schierenberg
BACKGROUND: The free-living nematode Diploscapter coronatus is the closest known relative of Caenorhabditis elegans with parthenogenetic reproduction. It shows several developmental idiosyncracies, for example concerning the mode of reproduction, embryonic axis formation and early cleavage pattern (Lahl et al. in Int J Dev Biol 50:393-397, 2006). Our recent genome analysis (Hiraki et al. in BMC Genomics 18:478, 2017) provides a solid foundation to better understand the molecular basis of developmental idiosyncrasies in this species in an evolutionary context by comparison with selected other nematodes...
2017: EvoDevo
https://www.readbyqxmd.com/read/29074739/genome-wide-discovery-of-active-regulatory-elements-and-transcription-factor-footprints-in-caenorhabditis-elegans-using-dnase-seq
#13
Paul Sternberg, Margaret Ho, Porfirio Quintero-Cadena
Deep sequencing of size-selected DNase I-treated chromatin (DNase-seq) allows high resolution measurement of chromatin accessibility to DNase I cleavage, permitting identification of de novo active cis-regulatory modules (CRMs) and individual transcription factor (TF) binding sites. We adapted DNase-seq to nuclei isolated from C. elegans embryos and L1 arrest larvae to generate high-resolution maps of TF binding. Over half of embryonic DNase I hypersensitive sites (DHSs) were annotated as noncoding, with 24% in intergenic, 12% in promoters and 28% in introns, with similar statistics observed in L1 arrest larvae...
October 26, 2017: Genome Research
https://www.readbyqxmd.com/read/29058002/a-microfluidic-microinjector-for-toxicological-and-developmental-studies-in-drosophila-embryos
#14
Reza Ghaemi, Pouya Arefi, Ana Stosic, Meryl Acker, Qanber Raza, J Roger Jacobs, Ponnambalam Ravi Selvaganapathy
Microinjection is an established and reliable method to deliver biological reagents such as transgenic constructs and drugs, to specific locations inside organisms such as the Drosophila embryo and C. elegans worm. In this paper, a simple compliant mechanism based PDMS-microinjection system has been demonstrated. Unlike conventional microinjectors, this unique system could allow one to precisely insert a long taper microneedle laterally and at various positions inside the length of the Drosophila embryo (up to 250 μm) with the resolution of 5 μm...
November 7, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/29049853/nutrient-driven-o-glcnac-in-proteostasis-and-neurodegeneration
#15
REVIEW
Ilhan Akan, Stephanie Olivier-Van Stichelen, Michelle R Bond, John A Hanover
Proteostasis is essential in the mammalian brain where post-mitotic cells must function for decades to maintain synaptic contacts and memory. The brain is dependent on glucose and other metabolites for proper function and is spared from metabolic deficits even during starvation. In this review, we outline how the nutrient sensitive nucleocytoplasmic posttranslational modification O-linked N-acetylglucosamine (O-GlcNAc) regulates protein homeostasis. The O-GlcNAc modification is highly abundant in the mammalian brain, and has been linked to proteopathies, including neurodegenerative diseases such as Alzheimer's, Parkinson's, and Huntington's...
October 19, 2017: Journal of Neurochemistry
https://www.readbyqxmd.com/read/29048526/comparative-transcriptomics-of-steinernema-and-caenorhabditis-single-embryos-reveals-orthologous-gene-expression-convergence-during-late-embryogenesis
#16
Marissa Macchietto, Dristi Angdembey, Negar Heidarpour, Lorrayne Serra, Bryan Rodriguez, Nicole El-Ali, Ali Mortazavi
Cells express distinct sets of genes in a precise spatio-temporal manner during embryonic development. There is a wealth of information on the deterministic embryonic development of Caenorhabditis elegans, but much less is known about embryonic development in nematodes from other taxa, especially at the molecular level. We are interested in insect pathogenic nematodes from the genus Steinernema as models of parasitism and symbiosis as well as a satellite model for evolution in comparison to C. elegans. To explore gene expression differences across taxa, we sequenced the transcriptomes of single embryos of two Steinernema species and two Caenorhabditis species at 11 stages during embryonic development and found several interesting features...
October 1, 2017: Genome Biology and Evolution
https://www.readbyqxmd.com/read/29043078/a-gap-that-divides
#17
REVIEW
Angika Basant, Michael Glotzer
Cytokinesis in metazoan cells is mediated by an actomyosin-based contractile ring that assembles in response to activation of the small GTPase RhoA. The guanine nucleotide exchange factor that activates RhoA during cytokinesis, ECT-2, is highly regulated. In most metazoan cells, with the notable exception of the early Caenorhabditis elegans embryo, RhoA activation and furrow ingression require the centralspindlin complex. This exception is due to the existence of a parallel pathway for RhoA activation in C...
2017: F1000Research
https://www.readbyqxmd.com/read/29030842/microrna-detection-by-whole-mount-in-situ-hybridization-in-c-elegans
#18
Yoshiki Andachi, Yuji Kohara
MicroRNAs (miRNAs) loaded on argonaute proteins guide RNA-induced silencing complexes to target mRNAs. An excellent method to decipher the spatiotemporal expression patterns of miRNAs is whole-mount in situ hybridization (WISH), which has been successfully used in vertebrate embryos but still remains unavailable for many animal species. Here, we describe a WISH method for miRNA detection in Caenorhabditis elegans at both embryonic and post-embryonic stages. Strategies devised for detection include fixation of animals with carbodiimide at a high temperature and subsequent partial digestion of the fixed animals with an extremely high concentration of proteinase...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29030390/apc-c-fzr-1-controls-sas-5-levels-to-regulate-centrosome-duplication-in-caenorhabditis-elegans
#19
Jeffrey C Medley, Lauren E DeMeyer, Megan M Kabara, Mi Hye Song
As the primary microtubule-organizing center, centrosomes play a key role in establishing mitotic bipolar spindles that secure correct transmission of genomic content. For the fidelity of cell division, centrosome number must be strictly controlled by duplicating only once per cell cycle. Proper levels of centrosome proteins are shown to be critical for normal centrosome number and function. Overexpressing core centrosome factors leads to extra centrosomes, while depleting these factors results in centrosome duplication failure...
October 13, 2017: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/28994576/combined-computed-nanotomography-and-nanoscopic-x-ray-fluorescence-imaging-of-cobalt-nanoparticles-in-caenorhabditis-elegans
#20
Simone Cagno, Dag Anders Brede, Gert Nuyts, Frederik Vanmeert, Alexandra Pacureanu, Remi Tucoulou, Peter Cloetens, Gerald Falkenberg, Koen Janssens, Brit Salbu, Ole Christian Lind
Synchrotron radiation phase-contrast computed nanotomography (nano-CT) and two- and three-dimensional (2D and 3D) nanoscopic X-ray fluorescence (nano-XRF) were used to investigate the internal distribution of engineered cobalt nanoparticles (Co NPs) in exposed individuals of the nematode Caenorhabditis elegans. Whole nematodes and selected tissues and organs were 3D-rendered: anatomical 3D renderings with 50 nm voxel size enabled the visualization of spherical nanoparticle aggregates with size up to 200 nm within intact C...
November 7, 2017: Analytical Chemistry
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