An analysis of the response to gut induction in the C. elegans embryo

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1227-1236 ◽  
Author(s):  
B. Goldstein
Keyword(s):  
Body Wall ◽  
Cell Lineage ◽  
Muscle Cells ◽  
The Other ◽  
Cell Stage ◽  
Pharyngeal Muscle ◽  
Cell Cycles ◽  
Sister Cell ◽  
C Elegans ◽  
Cell Diversity

Establishment of the gut founder cell (E) in C. elegans involves an interaction between the P2 and the EMS cell at the four cell stage. Here I show that the fate of only one daughter of EMS, the E cell, is affected by this induction. In the absence of the P2-EMS interaction, both E and its sister cell, MS, produce pharyngeal muscle cells and body wall muscle cells, much as MS normally does. By cell manipulations and inhibitor studies, I show first that EMS loses the competence to respond before it divides even once, but P2 presents an inducing signal for at least three cell cycles. Second, induction on one side of the EMS cell usually blocks the other side from responding to a second P2-derived signal. Third, microfilaments and microtubules may be required near the time of the interaction for subsequent gut differentiation. Lastly, cell manipulations in pie-1 mutant embryos, in which the P2 cell is transformed to an EMS-like fate and produces a gut cell lineage, revealed that gut fate is segregated to one of P2's daughters cell-autonomously. The results contrast with previous results from similar experiments on the response to other inductions, and suggest that this induction may generate cell diversity by a different mechanism.

When and how does cell division order influence cell allocation to the inner cell mass of the mouse blastocyst?

Development ◽  
1987 ◽  
Vol 100 (2) ◽  
pp. 325-332
Author(s):  
C.L. Garbutt ◽  
M.H. Johnson ◽  
M.A. George
Keyword(s):  
Cell Division ◽  
Cell Population ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
The Other ◽  
Mouse Blastocyst ◽  
Short Term ◽  
Cell Stage ◽  
Cell Embryo

Aggregate 8-cell embryos were constructed from four 2/8 pairs of blastomeres, one of which was marked with a short-term cell lineage marker and was also either 4 h older (derived from an early-dividing 4-cell) or 4 h younger (derived from a late-dividing 4-cell) than the other three pairs. The aggregate embryos were cultured to the 16-cell stage, at which time a second marker was used to label the outside cell population. The embryos were then disaggregated and each cell was examined to determine its labelling pattern. From this analysis, we calculated the relative contributions to the inside cell population of the 16-cell embryo of older and younger cells. Older cells were found to contribute preferentially. However, if the construction of the aggregate 8-cell embryo was delayed until each of the contributing 2/8 cell pairs had undergone intercellular flattening and then had been exposed to medium low in calcium to reverse this flattening immediately prior to aggregation, the advantage possessed by the older cells was lost. These results support the suggestion that older cells derived from early-dividing 4-cell blastomeres contribute preferentially to the inner cell mass as a result of being early-flattening cells.

Establishment of gut fate in the E lineage of C. elegans: the roles of lineage-dependent mechanisms and cell interactions

Development ◽  
1993 ◽  
Vol 118 (4) ◽  
pp. 1267-1277 ◽  
Author(s):  
B. Goldstein
Keyword(s):  
Cell Interactions ◽  
The Other ◽  
Cell Contact ◽  
Cell Stage ◽  
C Elegans ◽  
Daughter Cells ◽  
Intact Embryo ◽  
A Cell ◽  
Random Position ◽  
Recombination Experiments

The gut of C. elegans derives from all the progeny of the E blastomere, a cell of the eight cell stage. Previous work has shown that gut specification requires an induction during the four cell stage (Goldstein, B. (1992) Nature 357, 255–257). Blastomere isolation and recombination experiments were done to determine which parts of the embryo can respond to gut induction. Normally only the posterior side of the EMS blastomere contacts the inducing cell, P2. When P2 was instead placed in a random position on an isolated EMS, gut consistently differentiated from the daughter of EMS contacting P2, indicating that any side of EMS can respond to gut induction. Additionally, moving P2 around to the opposite side of EMS in an otherwise intact embryo caused EMS's two daughter cells to switch lineage timings, and gut to differentiate from the descendents of what normally would be the MS blastomere. The other cells of the four cell stage, ABa, ABp, and P2, did not form gut when placed in contact with the inducer. To determine whether any other inductions are involved in gut specification, timed blastomere isolations were done at the two and eight cell stages. In the absence of cell contact at the two cell stage, segregation of gut fate proceeded normally at both the two and four cell stages. Gut fate also segregated properly in the absence of cell contact at the eight cell stage. A model is presented for the roles of lineage-dependent mechanisms and cell interactions in establishing gut fate in the E lineage.

Features of cell lineage in preimplantation mouse development

Development ◽  
1978 ◽  
Vol 48 (1) ◽  
pp. 53-72
Author(s):  
C. F. Graham ◽  
Z. A. Deussen
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
The Other ◽  
Mouse Development ◽  
Cell Stage ◽  
Daughter Cells ◽  
Preimplantation Mouse ◽  
Peripheral Cells

The cell lineage of the mouse was studied from the 2-cell stage to the blastocyst. Lineage to the 8-cell stage was followed under the microscope. Each cell from the 2-cell stage divided to form two daughter cells which remained attached. Subsequently, these two daughters each produced two descendants; one of these descendants regularly lay deep in the structure of the embryo while the other was peripheral. Lineage to the blastocyst was followed by injecting oil drops into cells at the 8-cell stage, and then following the segregation of these drops into the inner cell mass and trophectoderm. Between the 8-cell stage and the blastocyst, the deep cells contributed more frequently to the inner cell mass than did the peripheral cells.

scholarly journals Assembly of body wall muscle and muscle cell attachment structures in Caenorhabditis elegans

1994 ◽  
Vol 124 (4) ◽  
pp. 491-506 ◽  
Author(s):  
MC Hresko ◽  
BD Williams ◽  
RH Waterston
Keyword(s):  
Spatial Organization ◽  
Body Wall ◽  
Cell Attachment ◽  
Temporal Distribution ◽  
Muscle Cells ◽  
Structural Elements ◽  
C Elegans ◽  
Attachment Structures ◽  
Outer Covering ◽  
Dividing Cells

C. Elegans has four muscle quadrants that are used for locomotion. Contraction is converted to locomotion because muscle cells are anchored to the cuticle (the outer covering of the worm) by a specialized basement membrane and hemidesmosome structures in the hypodermis (a cellular syncytium that covers the worm and secretes the cuticle). To study muscle assembly, we have used antibodies to determine the spatial and temporal distribution of muscle and attachment structure components in wild-type and mutant C. elegans embryos. Myofibrillar components are first observed diffusely distributed in the muscle cells, and are expressed in some dividing cells. Later, the components accumulate at the membrane adjacent to the hypodermis where the sarcomeres will form, showing that the cells have become polarized. Assembly of muscle attachment structures is spatially and temporally coordinated with muscle assembly suggesting that important developmental signals may be passed between muscle and hypodermal cells. Analysis of embryos homozygous for mutations that affect muscle assembly show that muscle components closer to the membrane than the affected protein assemble quite well, while those further from the membrane do not. Our results suggest a model where lattice assembly is initiated at the membrane and the spatial organization of the structural elements of the muscle is dictated by membrane proximal events, not by the filament components themselves.

scholarly journals Microinjection into the Caenorhabditis elegans embryo using an uncoated glass needle enables cell lineage visualization and reveals cell-non-autonomous adhesion control

2018 ◽  
Author(s):  
Yohei Kikuchi ◽  
Akatsuki Kimura
Keyword(s):  
Caenorhabditis Elegans ◽  
Cell Biology ◽  
Cell Lineage ◽  
Special Equipment ◽  
Cell Stage ◽  
C Elegans ◽  
Carbon Coated ◽  
A Cell ◽  
Glass Needle ◽  
Time Specific

AbstractMicroinjection is a useful method in cell biology, with which exogenous substances are introduced into a cell in a location- and time-specific manner. The Caenorhabditis elegans embryo is an important model system for cell and developmental biology. Applying microinjection to the C. elegans embryo had been difficult due to the rigid eggshell surrounding the embryo. In 2013, microinjection method using a carbon-coated quartz needle for the C. elegans embryo was reported. To prepare the needle, unfortunately, special equipment is required and thus a limited number of researchers can use this method. In this study, we established a method for the microinjection of drugs, dyes, and microbeads into the C. elegans embryo using an uncoated glass needle that can be produced in a general laboratory. This method enabled us to easily detect cell lineage up to adult stages by injecting a fluorescent dye into a blastomere. We also found a cell-non-autonomous control mechanism of cell adhesion; specifically, the injection of an actin inhibitor into one cell at the 2-cell stage enhanced adhesion between daughter cells of the other cell. Our microinjection method is expected to be used for broad studies and could facilitate various discoveries using C. elegans.

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’.

Early transcription in Caenorhabditis elegans embryos

Development ◽  
1994 ◽  
Vol 120 (2) ◽  
pp. 443-451 ◽  
Author(s):  
L.G. Edgar ◽  
N. Wolf ◽  
W.B. Wood
Keyword(s):  
Cell Division ◽  
Caenorhabditis Elegans ◽  
Cell Stage ◽  
Cell Cycles ◽  
Cell Lineages ◽  
C Elegans ◽  
Early Transcription ◽  
Alpha Amanitin ◽  
Autoradiographic Detection ◽  
Specific Timing

We have analysed early transcription in devitellinized, cultured embryos of the nematode Caenorhabditis elegans by two methods: measurement of [32P]UTP uptake into TCA-precipitable material and autoradiographic detection of [3H]UTP labelling both in the presence and absence of alpha-amanitin. RNA synthesis was first detected at the 8- to 12-cell stage, and alpha-amanitin sensitivity also appeared at this time, during the cleavages establishing the major founder cell lineages. The requirements for maternally supplied versus embryonically produced gene products in early embryogenesis were examined in the same culture system by observing the effects of alpha-amanitin on cell division and the early stereotyped lineage patterns. In the presence of high levels of alpha-amanitin added at varying times from two cells onward, cell division continued until approximately the 100-cell stage and then stopped during a single round of cell division. The characteristic unequal early cleavages, orientation of cleavage planes and lineage-specific timing of early divisions were unaffected by alpha-amanitin in embryos up to 87 cells. These results indicate that embryonic transcription starts well before gastrulation in C. elegans embryos, but that although embryonic transcripts may have important early functions, maternal products can support at least the mechanics of the first 6 to 7 cell cycles.

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 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 Cleavage, gastrulation, and germ disc formation of the amphipod Orchestia cavimana (Crustacea, Malacostraca, Peracarida)

2002 ◽  
Vol 71 (1-3) ◽  
pp. 9-28 ◽  
Author(s):  
Gerhard Scholtz ◽  
Carsten Wolff
Keyword(s):  
Embryonic Development ◽  
Video Recording ◽  
Cell Lineage ◽  
Embryonic Cell ◽  
Lineage Tracing ◽  
The Other ◽  
Cell Stage ◽  
Characteristic Type ◽  
Orchestia Cavimana ◽  
Disc Formation

Investigations of amphipod embryonic development have a long tradition. However, many aspects of amphipod embryology are still controversial. These concern, among others, the nature of the cleavage, the origin of the germ disc, and the mode of gastrulation. On the other hand, amphipods show the same characteristic type of invariant cell division pattern in the germ band as other malacostracans. Since amphipods seem to undergo a stereotyped pattern of early cleavage they are highly interesting for our understanding of the evolution of arthropod development. In this paper, we describe the cleavage pattern of the amphipod crustacean Orchestia cavimana from the zygote to gastrulation and the formation of the germ disc using direct observation, scanning electron microscopy, histology, video recording, and lineage tracing with a vital dye. The early development follows the mode of a total, radial, unequal cleavage with a determinate stereotyped pattern. A small transient blastocoel is formed. The 8-cell stage is characterised by 4 micromeres and 4 macromeres. One quadrant is smaller than the others. There are two kinds of eggs that show a mirror handed image. The 16-cell stage is the last regular stage after which the blastomeres divide highly asynchronously. The germ disc is formed by the descendants of the macromeres and some micromere derivatives. The other micromeres constitute the extra-embryonic region. Migration of macromere descendants is involved in germ disc formation accompanied by the extrusion of the yolk. During this process some vitellophages are formed. The gastrulation sensu stricto is initiated by the micromere derivatives of the smallest quadrant at the anterior of the forming germ disc. A true blastopore occurs which involves an invagination and the immigration of cells. Our data help to correct erroneous interpretations of former students of amphipod development. We can show that many characters of amphipod embryonic development are apomorphic supporting amphipod monophyly. With the present investigation we contribute to a complete understanding of the embryonic cell lineage of amphipods from the egg to segment formation and organogenesis.

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