Immunohistochemistry

Duerr, Janet S.

doi:10.1895/wormbook.1.105.1

Cited by 126 publications

(104 citation statements)

References 13 publications

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“…Antibody staining of dissected gonads was carried out as described (15), and staining of whole animals was carried out as described (43). See SI Materials and Methods for protocols.…”

Section: Methodsmentioning

confidence: 99%

Discovery of two GLP-1/Notch target genes that account for the role of GLP-1/Notch signaling in stem cell maintenance

Kershner

Shin

Hansen

et al. 2014

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

102

241

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A stem cell’s immediate microenvironment creates an essential “niche” to maintain stem cell self-renewal. Many niches and their intercellular signaling pathways are known, but for the most part, the key downstream targets of niche signaling remain elusive. Here, we report the discovery of two GLP-1/Notch target genes, lst-1 (lateral signaling target) and sygl-1 (synthetic Glp), that function redundantly to maintain germ-line stem cells (GSCs) in the nematode Caenorhabditis elegans. Whereas lst-1 and sygl-1 single mutants appear normal, lst-1 sygl-1 double mutants are phenotypically indistinguishable from glp-1/Notch mutants. Multiple lines of evidence demonstrate that GLP-1/Notch signaling activates lst-1 and sygl-1 expression in GSCs within the niche. Therefore, these two genes fully account for the role of GLP-1/Notch signaling in GSC maintenance. Importantly, lst-1 and sygl-1 are not required for GLP-1/Notch signaling per se. We conclude that lst-1 and sygl-1 forge a critical link between Notch signaling and GSC maintenance.

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“…Antibody staining of dissected gonads was carried out as described (15), and staining of whole animals was carried out as described (43). See SI Materials and Methods for protocols.…”

Section: Methodsmentioning

confidence: 99%

Discovery of two GLP-1/Notch target genes that account for the role of GLP-1/Notch signaling in stem cell maintenance

Kershner

Shin

Hansen

et al. 2014

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

102

241

View full text Add to dashboard Cite

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“…Worms were kept well fed to prevent conflation of aging traits with starvation, and measurements were made on Day 9 after hatching. For fixation experiments, worms were treated with 4% paraformaldehyde overnight before imaging (32).…”

Section: Methodsmentioning

confidence: 99%

Worms under Pressure: Bulk Mechanical Properties of C. elegans Are Independent of the Cuticle

Gilpin

Uppaluri

Brangwynne

2015

Biophysical Journal

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The mechanical properties of cells and tissues play a well-known role in physiology and disease. The model organism Caenorhabditis elegans exhibits mechanical properties that are still poorly understood, but are thought to be dominated by its collagen-rich outer cuticle. To our knowledge, we use a novel microfluidic technique to reveal that the worm responds linearly to low applied hydrostatic stress, exhibiting a volumetric compression with a bulk modulus, k ¼ 140 5 20 kPa; applying negative pressures leads to volumetric expansion of the worm, with a similar bulk modulus. Surprisingly, however, we find that a variety of collagen mutants and pharmacological perturbations targeting the cuticle do not impact the bulk modulus. Moreover, the worm exhibits dramatic stiffening at higher stresses-behavior that is also independent of the cuticle. The stress-strain curves for all conditions can be scaled onto a master equation, suggesting that C. elegans exhibits a universal elastic response dominated by the mechanics of pressurized internal organs.

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“…For immunohistochemical analysis, antibodies were obtained from external sources or raised against on-site produced Histagged proteins. The mf were fixed, permeabilized, and treated by an adaptation of the tube protocol for C. elegans (39). Controls include observation of mf with omission of primary antibody and, for on-site produced proteins, with peptide-or protein-adsorbed primary antibodies.…”

Section: Methodsmentioning

confidence: 99%

Ivermectin disrupts the function of the excretory-secretory apparatus in microfilariae of Brugia malayi

Moreno

Nabhan

Solomon

et al. 2010

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

147

141

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Ivermectin (IVM) is a broad-spectrum anthelmintic used in filariasis control programs. By binding to nematode glutamate-gated chloride channels (GluCls), IVM disrupts neurotransmission processes regulated by GluCl activity. IVM treatment of filarial infections is characterized by an initial dramatic drop in the levels of circulating microfilariae, followed by long-term suppression of their production, but the drug has little direct effect on microfilariae in culture at pharmacologically relevant concentrations. We localized Brugia malayi GluCl expression solely in a muscle structure that surrounds the microfilarial excretory-secretory (ES) vesicle, which suggests that protein release from the ES vesicle is regulated by GluCl activity. Consistent with this hypothesis, exposure to IVM in vitro decreased the amount of protein released from microfilariae. To better understand the scope of IVM effects on protein release by the parasite, three different expression patterns were identified from immunolocalization assays on a representative group of five microfilarial ES products. Patterns of expression suggest that the ES apparatus is the main source of regulated ES product release from microfilariae, as it is the only compartment that appears to be under neuromuscular control. Our results show that IVM treatment of microfilariae results in a marked reduction of protein release from the ES apparatus. Under in vivo conditions, the rapid microfilarial clearance induced by IVM treatment is proposed to result from suppression of the ability of the parasite to secrete proteins that enable evasion of the host immune system. filarial nematode | macrocyclic lactone | glutamate-gated chloride channels

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Immunohistochemistry

Cited by 126 publications

References 13 publications

Discovery of two GLP-1/Notch target genes that account for the role of GLP-1/Notch signaling in stem cell maintenance

Discovery of two GLP-1/Notch target genes that account for the role of GLP-1/Notch signaling in stem cell maintenance

Worms under Pressure: Bulk Mechanical Properties of C. elegans Are Independent of the Cuticle

Ivermectin disrupts the function of the excretory-secretory apparatus in microfilariae of Brugia malayi

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