scholarly journals Type IV Collagen Is Detectable in Most, but Not All, Basement Membranes of Caenorhabditis elegans and Assembles on Tissues That Do Not Express It

1997 ◽  
Vol 137 (5) ◽  
pp. 1171-1183 ◽  
Author(s):  
Patricia L. Graham ◽  
Jeffrey J. Johnson ◽  
Shaoru Wang ◽  
Marion H. Sibley ◽  
Malini C. Gupta ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Body Wall ◽  
Type Iv Collagen ◽  
Fluorescent Protein ◽  
Muscle Cells ◽  
Basement Membranes ◽  
Body Wall Muscle ◽  
Collagen Molecule ◽  
C Elegans ◽  
Type Iv

Type IV collagen in Caenorhabditis elegans is produced by two essential genes, emb-9 and let-2, which encode α1- and α2-like chains, respectively. The distribution of EMB-9 and LET-2 chains has been characterized using chain-specific antisera. The chains colocalize, suggesting that they may function in a single heterotrimeric collagen molecule. Type IV collagen is detected in all basement membranes except those on the pseudocoelomic face of body wall muscle and on the regions of the hypodermis between body wall muscle quadrants, indicating that there are major structural differences between some basement membranes in C. elegans. Using lacZ/green fluorescent protein (GFP) reporter constructs, both type IV collagen genes were shown to be expressed in the same cells, primarily body wall muscles, and some somatic cells of the gonad. Although the pharynx and intestine are covered with basement membranes that contain type IV collagen, these tissues do not express either type IV collagen gene. Using an epitope-tagged emb-9 construct, we show that type IV collagen made in body wall muscle cells can assemble into the pharyngeal, intestinal, and gonadal basement membranes. Additionally, we show that expression of functional type IV collagen only in body wall muscle cells is sufficient for C. elegans to complete development and be partially fertile. Since type IV collagen secreted from muscle cells only assembles into some of the basement membranes that it has access to, there must be a mechanism regulating its assembly. We propose that interaction with a cell surface–associated molecule(s) is required to facilitate type IV collagen assembly.

scholarly journals Three-dimensional simulation of the Caenorhabditis elegans body and muscle cells in liquid and gel environments for behavioural analysis

2018 ◽  
Vol 373 (1758) ◽  
pp. 20170376 ◽  
Author(s):  
Andrey Palyanov ◽  
Sergey Khayrulin ◽  
Stephen D. Larson
Keyword(s):  
Caenorhabditis Elegans ◽  
Muscle Activation ◽  
Body Wall ◽  
Particle System ◽  
Three Dimensional ◽  
Muscle Cells ◽  
The Body ◽  
C Elegans ◽  
Muscle Activation Patterns ◽  
Simulation Engine

To better understand how a nervous system controls the movements of an organism, we have created a three-dimensional computational biomechanical model of the Caenorhabditis elegans body based on real anatomical structure. The body model is created with a particle system–based simulation engine known as Sibernetic, which implements the smoothed particle–hydrodynamics algorithm. The model includes an elastic body-wall cuticle subject to hydrostatic pressure. This cuticle is then driven by body-wall muscle cells that contract and relax, whose positions and shape are mapped from C. elegans anatomy, and determined from light microscopy and electron micrograph data. We show that by using different muscle activation patterns, this model is capable of producing C. elegans -like behaviours, including crawling and swimming locomotion in environments with different viscosities, while fitting multiple additional known biomechanical properties of the animal.  This article is part of a discussion meeting issue ‘Connectome to behaviour: modelling C. elegans at cellular resolution’.

MEC-8 regulates alternative splicing ofunc-52transcripts inC. eleganshypodermal cells

Development ◽  
2002 ◽  
Vol 129 (21) ◽  
pp. 4999-5008 ◽  
Author(s):  
Caroline A. Spike ◽  
Andrew G. Davies ◽  
Jocelyn E. Shaw ◽  
Robert K. Herman
Keyword(s):  
Alternative Splicing ◽  
Body Wall ◽  
Muscle Development ◽  
Rna Binding ◽  
Fluorescent Protein ◽  
Body Wall Muscle ◽  
Larval Body ◽  
C Elegans ◽  
Green Fluorescent ◽  
Mutant Phenotypes

Previous work has shown that C. elegans MEC-8 is a putative RNA-binding protein that promotes specific alternative splices ofunc-52 transcripts. unc-52 encodes homologs of mammalian perlecan that are located extracellularly between muscle and hypodermis and are essential for muscle development in both embryos and larvae. We show that MEC-8 is a nuclear protein found in hypodermis at most stages of development and not in most late embryonic or larval body-wall muscle. We have also found that overexpression of MEC-8 in hypodermis but not muscle can suppress certainunc-52 mutant phenotypes. These are unexpected results because it has been proposed that UNC-52 is produced exclusively by muscle. We have constructed various tissue-specific unc-52 minigenes fused to a gene for green fluorescent protein that have allowed us to monitor tissue-specificmec-8-dependent alternative splicing; we show that mec-8must be expressed in the same cell type as the unc-52 minigene in order to regulate its expression, supporting the view that MEC-8 acts directly on unc-52 transcripts and that UNC-52 must be synthesized primarily by the hypodermis. Indeed, our analysis of unc-52 genetic mosaics has shown that the focus of unc-52 action is not in body-wall muscle but most likely is in hypodermis.

Differential assembly of alpha- and gamma-filagenins into thick filaments in Caenorhabditis elegans

2000 ◽  
Vol 113 (22) ◽  
pp. 4001-4012 ◽  
Author(s):  
F. Liu ◽  
I. Ortiz ◽  
A. Hutagalung ◽  
C.C. Bauer ◽  
R.G. Cook ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Immunoelectron Microscopy ◽  
Body Wall ◽  
Developmental Stages ◽  
Body Wall Muscle ◽  
Developmentally Regulated ◽  
C Elegans ◽  
Novel Proteins ◽  
Thick Filaments ◽  
Associated Proteins

Muscle thick filaments are highly organized supramolecular assemblies of myosin and associated proteins with lengths, diameters and flexural rigidities characteristic of their source. The cores of body wall muscle thick filaments of the nematode Caenorhabditis elegans are tubular structures of paramyosin sub-filaments coupled by filagenins and have been proposed to serve as templates for the assembly of native thick filaments. We have characterized alpha- and gamma-filagenins, two novel proteins of the cores with calculated molecular masses of 30,043 and 19,601 and isoelectric points of 10.52 and 11.49, respectively. Western blot and immunoelectron microscopy using affinity-purified antibodies confirmed that the two proteins are core components. Immunoelectron microscopy of the cores revealed that they assemble with different periodicities. Immunofluorescence microscopy showed that alpha-filagenin is localized in the medial regions of the A-bands of body wall muscle cells whereas gamma-filagenin is localized in the flanking regions, and that alpha-filagenin is expressed in 1.5-twofold embryos while gamma-filagenin becomes detectable only in late vermiform embryos. The expression of both proteins continues throughout later stages of development. C. elegans body wall muscle thick filaments of these developmental stages have distinct lengths. Our results suggest that the differential assembly of alpha- and gamma-filagenins into thick filaments of distinct lengths may be developmentally regulated.

scholarly journals Domain and basement membrane specificity of a monoclonal antibody against chicken type IV collagen.

1982 ◽  
Vol 95 (2) ◽  
pp. 641-647 ◽  
Author(s):  
J M Fitch ◽  
E Gibney ◽  
R D Sanderson ◽  
R Mayne ◽  
T F Linsenmayer
Keyword(s):  
Monoclonal Antibody ◽  
Basement Membrane ◽  
Type Iv Collagen ◽  
Helical Structure ◽  
Immunohistochemical Localization ◽  
Basement Membranes ◽  
Optical Rotatory Dispersion ◽  
Collagen Molecule ◽  
Type Iv ◽  
Triple Helical Domain

A monoclonal antibody, IV-IA8, generated against chicken type IV collagen has been characterized and shown to bind specifically to a conformational-dependent site within a major, triple helical domain of the type IV molecule. Immunohistochemical localization of the antigenic determinant with IV-IA8 revealed that the basement membranes of a variety of chick tissues were stained but that the basement membrane of the corneal epithelium showed little, if any, staining. Thus, basement membranes may differ in their content of type IV collagen, or in the way in which it is assembled. The specificity of the antibody was determined by inhibition ELISA using purified collagen types I-V and three purified molecular domains of chick type IV collagen ([F1]2F2, F3, and 7S) as inhibitors. Only unfractionated type IV collagen and the (F1)2F2 domain bound the antibody. Antibody binding was destroyed by thermal denaturation of the collagen, the loss occurring at a temperature similar to that at which previous optical rotatory dispersion studies had shown melting of the triple helical structure of (F1)2F2. Such domain-specific monoclonal antibodies should prove to be useful probes in studies involving immunological dissection of the type IV collagen molecule, its assembly within basement membranes, and changes in its distribution during normal development and in disease.

scholarly journals The L-type voltage-dependent Ca2+ channel EGL-19 controls body wall muscle function in Caenorhabditis elegans

2002 ◽  
Vol 159 (2) ◽  
pp. 337-348 ◽  
Author(s):  
Maëlle Jospin ◽  
Vincent Jacquemond ◽  
Marie-Christine Mariol ◽  
Laurent Ségalat ◽  
Bruno Allard
Keyword(s):  
Caenorhabditis Elegans ◽  
Body Wall ◽  
Muscle Cells ◽  
Membrane Depolarization ◽  
Cellular Level ◽  
Body Wall Muscle ◽  
K Channel ◽  
Channel Blockers ◽  
Patch Clamp Technique ◽  
Voltage Dependent

Caenorhabditis elegans is a powerful model system widely used to investigate the relationships between genes and complex behaviors like locomotion. However, physiological studies at the cellular level have been restricted by the difficulty to dissect this microscopic animal. Thus, little is known about the properties of body wall muscle cells used for locomotion. Using in situ patch clamp technique, we show that body wall muscle cells generate spontaneous spike potentials and develop graded action potentials in response to injection of positive current of increasing amplitude. In the presence of K+ channel blockers, membrane depolarization elicited Ca2+ currents inhibited by nifedipine and exhibiting Ca2+-dependent inactivation. Our results give evidence that the Ca2+ channel involved belongs to the L-type class and corresponds to EGL-19, a putative Ca2+ channel originally thought to be a member of this class on the basis of genomic data. Using Ca2+ fluorescence imaging on patch-clamped muscle cells, we demonstrate that the Ca2+ transients elicited by membrane depolarization are under the control of Ca2+ entry through L-type Ca2+ channels. In reduction of function egl-19 mutant muscle cells, Ca2+ currents displayed slower activation kinetics and provided a significantly smaller Ca2+ entry, whereas the threshold for Ca2+ transients was shifted toward positive membrane potentials.

scholarly journals Genetic identification, sequence, and alternative splicing of the Caenorhabditis elegans alpha 2(IV) collagen gene.

1993 ◽  
Vol 123 (1) ◽  
pp. 255-264 ◽  
Author(s):  
M H Sibley ◽  
J J Johnson ◽  
C C Mello ◽  
J M Kramer
Keyword(s):  
Amino Acid ◽  
Caenorhabditis Elegans ◽  
Type Iv Collagen ◽  
Collagen Gene ◽  
C Elegans ◽  
Type Iv ◽  
High Amino Acid ◽  
Alternatively Spliced ◽  
Exon 9 ◽  
Collagen Genes

The nematode Caenorhabditis elegans has two type IV collagen genes homologous to the mammalian alpha 1(IV) and alpha 2(IV) collagen genes. We demonstrate by transgenic rescue of mutant animals that the genetic locus encoding the C. elegans alpha 2(IV) collagen gene is let-2 on the X chromosome. The most severe effect of mutations in let-2 is temperature-sensitive embryonic lethality. The embryonic lethal phenotype is similar to that seen in animals with mutations in the alpha 1(IV) collagen gene, emb-9. The sequence of the entire C. elegans alpha 2(IV) collagen gene is presented. Comparisons with mammalian type IV collagen sequences show high amino acid sequence conservation in the C-terminal NCl domain and of crosslinking residues (Cys and Lys) in the N-terminal 7S domain. RT-PCR analysis shows that transcripts of the C. elegans alpha 2(IV) collagen gene are alternatively spliced. Transcripts contain one of two mutually exclusive exons, exon 9 or 10. These exons encode very similar products, differing primarily in the sequence of a 9-10 amino acid Gly-X-Y interruption. The expression of these alternatively spliced alpha 2(IV) collagen transcripts is developmentally regulated. In embryos over 90% of the alpha 2(IV) collagen mRNA contains exon 9, while larval and adult RNAs contain 80-90% exon 10. This shift in expression of alternative alpha 2(IV) collagen transcripts suggests that C. elegans embryos may require a different form of alpha 2(IV) collagen than do larvae and adults.

scholarly journals Muscle-specific functions of ryanodine receptor channels in Caenorhabditis elegans

1998 ◽  
Vol 111 (19) ◽  
pp. 2885-2895 ◽  
Author(s):  
E.B. Maryon ◽  
B. Saari ◽  
P. Anderson
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Ryanodine Receptor ◽  
Body Wall ◽  
Calcium Release ◽  
Muscle Cells ◽  
Body Wall Muscle ◽  
Apical Plasma Membrane ◽  
C Elegans ◽  
Terminal Bulb ◽  
Null Mutants

Ryanodine receptor channels regulate contraction of striated muscle by gating the release of calcium ions from the sarcoplasmic reticulum. Ryanodine receptors are expressed in excitable and non-excitable cells of numerous species, including the nematode C. elegans. Unlike vertebrates, which have at least three ryanodine receptor genes, C. elegans has a single gene encoded by the unc-68 locus. We show that unc-68 is expressed in most muscle cells, and that the phenotypic defects exhibited by unc-68 null mutants result from the loss of unc-68 function in pharyngeal and body-wall muscle cells. The loss of unc-68 function in the isthmus and terminal bulb muscles of the pharynx causes a reduction in growth rate and brood size. unc-68 null mutants exhibit defective pharyngeal pumping (feeding) and have abnormal vacuoles in the terminal bulb of the pharynx. unc-68 is required in body-wall muscle cells for normal motility. We show that UNC-68 is localized in body-wall muscle cells to flattened vesicular sacs positioned between the apical plasma membrane and the myofilament lattice. In unc-68 mutants, the vesicles are enlarged and densely stained. The flattened vesicles in body-wall muscle cells thus represent the C. elegans sarcoplasmic reticulum. Morphological and behavioral phenotypes of unc-68 mutants suggest that intracellular calcium release is not essential for excitation-contraction coupling in C. elegans.

scholarly journals α-Catulin CTN-1 is required for BK channel subcellular localization in C. elegans body-wall muscle cells

2010 ◽  
Vol 29 (18) ◽  
pp. 3184-3195 ◽  
Author(s):  
Bojun Chen ◽  
Ping Liu ◽  
Sijie J Wang ◽  
Qian Ge ◽  
Haiying Zhan ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Body Wall ◽  
Muscle Cells ◽  
Bk Channel ◽  
Body Wall Muscle ◽  
C Elegans

scholarly journals Genetic dissection of ion currents underlying all-or-none action potentials in C. elegans body-wall muscle cells

2010 ◽  
Vol 589 (1) ◽  
pp. 101-117 ◽  
Author(s):  
P. Liu ◽  
Q. Ge ◽  
B. Chen ◽  
L. Salkoff ◽  
M. I. Kotlikoff ◽  
...  
Keyword(s):  
Action Potentials ◽  
Body Wall ◽  
Muscle Cells ◽  
Body Wall Muscle ◽  
Genetic Dissection ◽  
Ion Currents ◽  
C Elegans

HSP43, a small heat-shock protein localized to specific cells of the vulva and spermatheca in the nematode Caenorhabditis elegans

2000 ◽  
Vol 349 (2) ◽  
pp. 409-412 ◽  
Author(s):  
Lily DING ◽  
E. M. Peter CANDIDO
Keyword(s):  
Caenorhabditis Elegans ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Body Wall ◽  
Small Heat Shock Protein ◽  
Muscle Cells ◽  
Body Wall Muscle ◽  
Nematode Caenorhabditis Elegans ◽  
Punctate Pattern

Heat-shock protein 43 (HSP43) of Caenorhabditis elegans is prominently expressed in the utse cell, which attaches the uterus to the hypodermis, the uv1 cells joining the vulva and the uterus, the spermathecal valve and junctions between cells of the spermathecal cage. In body-wall muscle, HSP43 forms a punctate pattern of circumferential lines, probably corresponding to regions where the hypodermis contacts the muscle cells.

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