87 Works

EOR-1 mediates non-cell autonomous regulation of abts-1 gene expression in HSNs

Yoichi Shinkai & Motomichi Doi
In the present study, we investigated whether abts-1 expression in HSNs is dependent on neuronal activities through synaptic transmission. abts-1 is expressed in HSNs from the L4 larval stage onward (Shinkai et al. 2018). To measure abts-1 expression levels in HSNs, we used transgenic worms carrying vsIs138 [Pabts-1b::gfp, lin15(+)]. Quantitative fluorescent measurements were performed as previously described (Shinkai et al. 2018). Since presynaptic inputs are important for the GABA functional switch, we investigated the effect...

Genetic mapping of EgfrL.3.1 in Drosophila melanogaster

Joyce Stamm, Gnanada S Joshi, MA Anderson, Katie Bussing, Colton Houchin, Amber C Elinsky, Jacob T Flyte, Nadine Husseini, Dominika Jarosz, Chelsea L Johnson, Abby F Johnson, Christina E Jones, Taj P Kooner, Daniel Myhre, Thomas N Rafaill, Sarah Sayed, Kirby W Swan, Jonathan Toma & Jacob D Kagey

Copulation defective mutants of C. elegans

Yvonne M. Hajdu-Cronin, Katharine S. Liu, Leslie Barber, Helen M. Chamberlin, William Boorstein & Paul W. Sternberg
​Description To identify genes involved in male copulatory behavior, we carried out an F2 clonal screen in a him-5 mutant background. We identified 20 mutations that affect male mating behavior without causing gross defects in morphology. ​ Male mating in C. elegans comprises at least five steps (Liu and Sternberg, 1995). (l) The male responds to the hermaphrodite by backing his tail along the length of the hermaphrodite, (2) he turns over or under her...

Application of the red-shifted channelrhodopsin Chrimson for the Caenorhabditis elegans cGAL bipartite system

Mengyi Cao, Cynthia Chai, Jonathan Liu & Paul W. Sternberg
Channelrhodopsins are light-gated ion channels that serve as photoreceptors in photosynthetic microbes and have been applied as crucial optogenetic tools in genetic model organisms. When expressed in animals, they enable light-inducible control of ionic membrane permeability, which directly manipulates the activity of neurons expressing the protein. The application of channelrhodopsin-based optogenetics is particularly powerful when used in conjugation with the cGAL (GAL4-UAS) bipartite system (Wang, 2017). The mating of neuron-specific GAL4 driver lines to new...

DVC interneuron cGAL driver in Caenorhabditis elegans

Jun Young Oh, Shahla Gharib, Jonathan Liu, Han Wang & Paul W Sternberg

Loss of eif-2alpha phosphorylation on S49 (mammalian S51) associated with the integrated stress response hastens development in C. elegans​

Jarod Rollins, Noah Lind & Aric N. Rogers
The gene eif-2alpha (Y37E3.10) in C. elegans encodes for the alpha subunit of eukaryotic translation initiation factor 2 (eIF2). The eIF2 trimer is required for delivering charged tRNAMet to the ribosome in a reaction requiring the hydrolysis of GTP to GDP (Sokabe et al., 2012). The phosphorylation of eif2-alpha on its 49th serine residue (Nukazuka et al., 2008) prevents replenishing GDP with GTP and thus reduces translation initiation by preventing delivery of tRNAMet. The phosphorylation...

GFPnovo2, a brighter GFP variant for in vivo labeling in C. elegans

Ardalan Hendi & Kota Mizumoto
Green fluorescent protein (GFP) is one of the most common fluorophores used to label cells and proteins in C. elegans. Previous artificial protein evolution strategy has generated a variant of eGFP, GFPnovo2, which is 3.3 times brighter than the original eGFP in the DT40 cell line (Arakawa et al, 2008). GFPnovo2 carries four mutations from the original eGFP (Y145F, V163A, S202T, L221V), and has the same excitation/emission wavelengths as the original eGFP. Here we compared...

Some effects of a chrysin bromide-derivative on GABA-A receptors and on Caenorhabditis elegans

José L Téllez-Arreola, Alejandro Valdez-Calderón, Simplicio González-Montiel, Ataúlfo Martinez-Torres & Adan Hernandez

daf-2 regeneration of mechanosensory neurons: integration

Zehra C Abay, Michelle Yu-Ying Wong & Brent Neumann
We previously established that a critical event for axonal fusion to occur is the exposure of PS ‘save-me’ signals after injury (Neumann et al., 2015), which increase with advancing age (Abay et al., 2017a). To determine if daf-2 is involved in this age-dependent modulation of PS exposure, we visualised and quantified the level of PS exposed after PLM axotomy using a secreted, tagged version of Annexin V (Neumann et al., 2015; Mapes et al., 2012)....

Mutations in KCNQ potassium channels cause pharyngeal pumping defects in C. elegans

Yoanne Clovis, Alexis Webb, Carl Turner & Bill Roberts
Pumps were stimulated with 10mM 5HT in M9 recorded as electropharyngeograms (EPGs) for 2 minutes in a NemaMetrix ScreenChip, and analyzed using NemAnalysis software (NemaMetrix). The null mutant strains kqt-1(aw3) and kqt-3 (aw1) were kindly donated by Dr. Aguan Wei (Wei et al., 2002) A) Pump frequency in kqt-1(aw3) animals was significantly lower than in N2s, while kqt-3(aw1) worms showed an increase in pump frequency (*p<0.05; ***p<0.01; 1-tailed Mann-Whitney U-test; n = 21-23 worms in...

2018 Update on Protein-Protein Interaction Data in WormBase

Jaehyoung Cho, Christian Grove, Kimberly Van Auken, Juancarlos Chan, Sibyl Gao & Paul W. Sternberg
Protein interaction is an important data type to understand the biological function of proteins involved in the interaction, and helps researchers to deduce the biological nature of unknown proteins from the well-characterized functions of their interaction partners. High-throughput studies, coupled with the aggregation of individual experiments, provides a global 'snapshot' of the protein interactions occurring at all levels of biological processes or circumstances. This snapshot of the interaction network, the interactome, is important to understand...

Extending the CRISPR toolbox for C. elegans: Cpf1 as an alternative gene editing system for AT-rich sequences

Annabel Ebbing, Peng Shang, Niels Geijsen & Hendrik C. Korswagen
The CRISPR/Cas9 system has become a powerful tool for genome-editing in C. elegans. Sequence specificity of the system is determined by a guide RNA which targets the Cas9 endonuclease to the genomic region of interest. In addition, Cas9 needs to interact with a so-called protospacer-adjacent motif (PAM) next to the target sequence. In case of Cas9 from Streptococcus pyogenes (SpCas9), the most commonly used Cas9, this sequence is NGG. Because the C. elegans genome sequence...

me98 is a new allele of rad-54

Baptiste Roelens, Karl A Zawadzki & Anne M Villeneuve
We isolated the me98 mutant in a genetic screen for C. elegans mutants with an altered number of GFP::COSA-1 foci, which mark the sites of crossovers in wild-type C. elegans germ cells (ROSU et al. 2013). After multiple rounds of outcrossing, we confirmed that the me98 mutant is defective in meiotic prophase as i) chromosomes in diakinesis oocytes appear partially decondensed and structurally compromised (Fig. 1A), ii) me98 fails to form the six GFP::COSA-1 foci...

GABA Receptor mutant gbb-1 accelerates morphological aging of GABA neurons in Caenorhabditis elegans

I. Dhillon, A. Momin & I. Chin-Sang
The aging of an organism is heterogeneous, that is to say, tissues and organs may age at unique rates compared to overall organism longevity. Inhibition of the insulin/insulin-like growth factor signaling (IIS), such as the daf-2 IIS receptor increases Caenorhabditis elegans longevity. The daf-2 pathway plays a key role in the regulation of metabolism and stress responses and as such, impacts the longevity of the organism (Uno and Nishida 2016). Evidence suggests that an increase...

wrt-2 expression oscillates during larval development

Ana G. Vera-Cruz, Siavash Amon, Sabih Rashid & Lesley T. MacNeil
wrt-2 encodes a hedgehog-like protein that is weakly required for molting (Kouns et al., 2011). As monitored using a transcriptional GFP reporter, wrt-2 is expressed in seam and hypodermal cells throughout development (Apock et al., 1999). However, wrt-2 transcript levels oscillate with a period consistent with the timing of the molting cycle (Hendriks et al., 2014). Because standard GFP reporters do not capture rapid changes in gene expression, we generated a wrt-2 transcriptional reporter driving...

Microfluidic EPG Recordings Show Striking Pharyngeal Pumping Phenotype in a C. elegans Alzheimer's Disease Model

Janis C Weeks, Kristen J Robinson, Benedicta Wanjeri, Philip F Copenhaver & William M Roberts
Strain CL4176 accumulates human amyloid-β1-42 in body wall muscles after animals are shifted from 15 to 25 oC, causing paralysis (Link et al. 2003). We temperature-shifted worms at early L4 and quantified pharyngeal pumping at 48 h using a microfluidic electropharyngeogram (EPG) recording platform (Lockery et al. 2012; Weeks et al. 2016). Recordings were made in M9 buffer with 10 mM serotonin and analyzed by automated pump-recognition software. At 48 h, control CL802 worms showed...

unc-42 regulates the expression of ASH terminal fate markers

Jordan F Wood & Denise M Ferkey
The genes encoding transcription factors that initiate the terminal differentiation programs within individual neurons have been termed “terminal selectors” (Hobert 2008). They are considered master regulators of neuronal identity because they integrate upstream lineage inputs to ultimately drive (directly and/or indirectly) the expression of all terminal differentiation features of a neuron (Hobert 2008; Bertrand and Hobert 2010). The paired-like homeodomain transcription factor UNC-42 has been proposed to function as the terminal selector in the developmental...

The Mapping of Drosophila melanogaster mutant A.4.4

Kayla L Bieser, Joyce Stamm, Ayala A Aldo, Suneil Bhaskara, Makayla Claiborne, Joselyn N Coronel Gómez, Ron Dean, Aaron Dowell, Evan Dowell, Mathew Eissa, Ahmad A Fawaz, Michael M Fouad-Meshriky, Dustin Godoy, Krista Gonzalez, Malak K Hachem, Malak F Hammoud, Anthony Huffman, Hunter Ingram, Alex B Jackman, Bibek Karki, Natalia Khalil, Houda Khalil, Tran Khanh Ha, Arjun Kharel, Izabell Kobylarz … & Jacob D Kagey
A novel Drosophila melanogaster mutant A.4.4 was isolated from a conditional Flp/FRT mosaic eye screen in the context of blocked apoptosis (Kagey et al., 2012). The ;FRT42D, Dark82 chromosome was used as a starting point for the EMS mutagenesis screen to screen for mutations that conferred a growth advantage in the environment of blocked apoptosis via the homozygous Dark82 allele (Akdemir et al., 2006). Mutants were screened for over-representation of mutant tissue (pigmented) as compared...

acdh-1 transcripts are expressed in the intestine

Dustin Updike
​Stellaris smFISH probes targeting acdh-1, a short-chain acyl-CoA dehydrogenase, are shown in red (Cal Fluor 610). DAPI/blue marks embryonic nuclei, and PGL-1::GFP shows the corresponding location of P granules surrounding germ cell nuclei (arrowheads, green). During embryogenesis (A), acdh-1 expression begins in the E cells (arrow, red). Expression continues in the developing intestine, shown in red, throughout embryogenesis. Intestinal expression of acdh-1 persists through larval development in the L1 (B) and L2 (C) stages. These...

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