Soma-germline asymmetry in the distributions of embryonic RNAs in Caenorhabditis elegans

Development ◽  
1994 ◽  
Vol 120 (10) ◽  
pp. 2823-2834 ◽  
Author(s):  
G. Seydoux ◽  
A. Fire
Keyword(s):  
Caenorhabditis Elegans ◽  
Somatic Cells ◽  
Early Embryogenesis ◽  
Messenger Rnas ◽  
Cell Stage ◽  
P Granules ◽  
C Elegans ◽  
Germline Cells ◽  
Hybridization Protocol

Early embryogenesis in Caenorhabditis elegans is characterized by a series of unequal cleavages that mark the stepwise separation of somatic and germ lineages. We have developed an in situ hybridization protocol to examine the localization of specific maternal and embryonically transcribed messenger RNAs during these early cleavages. We detected three classes of maternal RNAs: RNAs that are maintained in all cells, RNAs that are maintained in germline cells but are lost from somatic cells, and a population of RNAs that are associated with the germline-specific P granules. We observed embryonically transcribed RNAs in somatic cells as early as the 4-cell stage. These transcripts were not detected in germline cells. These observations suggest that mechanisms which distinguish between soma and germline cause asymmetries in mRNA stability and transcription within the first few cleavages of C. elegans embryogenesis.

scholarly journals Reevaluation of whether a soma–to–germ-line transformation extends lifespan in Caenorhabditis elegans

2016 ◽  
Vol 113 (13) ◽  
pp. 3591-3596 ◽  
Author(s):  
Andrew Kekūpa'a Knutson ◽  
Andreas Rechtsteiner ◽  
Susan Strome
Keyword(s):  
Young Adults ◽  
Caenorhabditis Elegans ◽  
Germ Line ◽  
Somatic Cells ◽  
Simultaneous Removal ◽  
Whole Genome ◽  
P Granules ◽  
C Elegans ◽  
Granule Proteins ◽  
Germ Line Transformation

The germ lineage is considered to be immortal. In the quest to extend lifespan, a possible strategy is to drive germ-line traits in somatic cells, to try to confer some of the germ lineage’s immortality on the somatic body. Notably, a study in Caenorhabditis elegans suggested that expression of germ-line genes in the somatic cells of long-lived daf-2 mutants confers some of daf-2’s long lifespan. Specifically, mRNAs encoding components of C. elegans germ granules (P granules) were up-regulated in daf-2 mutant worms, and knockdown of individual P-granule and other germ-line genes in daf-2 young adults modestly reduced their lifespan. We investigated the contribution of a germ-line program to daf-2’s long lifespan and also tested whether other mutants known to express germ-line genes in their somatic cells are long-lived. Our key findings are as follows. (i) We could not detect P-granule proteins in the somatic cells of daf-2 mutants by immunostaining or by expression of a P-granule transgene. (ii) Whole-genome transcript profiling of animals lacking a germ line revealed that germ-line transcripts are not up-regulated in the soma of daf-2 worms compared with the soma of control worms. (iii) Simultaneous removal of multiple P-granule proteins or the entire germ-line program from daf-2 worms did not reduce their lifespan. (iv) Several mutants that robustly express a broad spectrum of germ-line genes in their somatic cells are not long-lived. Together, our findings argue against the hypothesis that acquisition of a germ-cell program in somatic cells increases lifespan and contributes to daf-2’s long lifespan.

scholarly journals Assessment of asymmetric cell divisions in the early development of Caenorhabditis elegans

2017 ◽  
Author(s):  
Rolf Fickentscher ◽  
Matthias Weiss
Keyword(s):  
Caenorhabditis Elegans ◽  
Cell Size ◽  
Somatic Cells ◽  
Metaphase Plate ◽  
Early Embryogenesis ◽  
Asymmetric Cell Divisions ◽  
C Elegans ◽  
Light Sheet Microscopy ◽  
Cell Divisions ◽  
A Cell

AbstractAsymmetric cell divisions are of fundamental importance for developmental processes, e.g. for the generation of founder cells. Prime examples are asymmetric cell divisions in the P lineage during early embryogenesis of the model organism Caenorhabditis elegans. However, due to a lack of quantitative data it has remained unclear how frequent unequal daughter cell sizes emerge in the nematode’s early embryogenesis, and whether these originate from sterical or biochemical cues. Using quantitative light-sheet microscopy, we have found that about 40% of all cell divisions in C. elegans until gastrulation generate daughter cells with significantly different volumes. Removing the embryo’s rigid eggshell revealed asymmetric divisions in somatic cells to be primarily induced by steric effects. Division asymmetries in the germline remained unaltered and were correctly reproduced by a model based on a cell-size independent, eccentric displacement of the metaphase plate. Our data suggest asymmetric cell divisions to be essential for establishing important cell-cell interactions that eventually fuel a successful embryogenesis.Summary statementAbout 40% of all cell divisions in early C. elegans embryogenesis are found to be asymmetric. A cell-size independent displacement of the mitotic spindle explains division asymmetries in the germline whereas the confining eggshell induces asymmetries of somatic cells.

PES-1 is expressed during early embryogenesis in Caenorhabditis elegans and has homology to the fork head family of transcription factors

Development ◽  
1994 ◽  
Vol 120 (3) ◽  
pp. 505-514 ◽  
Author(s):  
I.A. Hope
Keyword(s):  
Transcription Factors ◽  
Caenorhabditis Elegans ◽  
Cell Lineage ◽  
Embryonic Cell ◽  
Early Embryogenesis ◽  
C Elegans ◽  
Promoter Trapping ◽  
Fork Head ◽  
Initiation Of Transcription ◽  
Beta Galactosidase

Promoter trapping has identified a gene, pes-1, which is expressed during C. elegans embryogenesis. The beta-galactosidase expression pattern, directed by the pes-1/lacZ fusion through which this gene was cloned, has been determined precisely in terms of the embryonic cell lineage and has three components. One component is in a subset of cells of the AB founder cell lineage during early embryogenesis, suggesting pes-1 may be regulated both by cell autonomous determinants and by intercellular signals. Analysis of cDNA suggests pes-1 has two sites for initiation of transcription and the two transcripts would encode related but distinct proteins. The predicted PES-1 proteins have homology to the fork head family of transcription factors and therefore may have important regulatory roles in early embryogenesis.

scholarly journals ATM Induces Cell Death with Autophagy in Response to H2O2 Specifically in Caenorhabditis elegans Nondividing Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Takahito Moriwaki ◽  
Akira Yamasaki ◽  
Qiu-Mei Zhang-Akiyama
Keyword(s):  
Cell Death ◽  
Caenorhabditis Elegans ◽  
Somatic Cells ◽  
Dead Cell ◽  
C Elegans ◽  
Cell Staining ◽  
Dna Damaging Agents ◽  
Gfp Reporter ◽  
Wide Range ◽  
Reporter Assays

Introduction. Ataxia-telangiectasia-mutated (ATM) kinase is a master regulator of the DNA damage response and is directly activated by reactive oxygen species (ROSs) in addition to DNA double-stranded breaks. However, the physiological function of the response to ROSs is not understood. Purpose. In the present study, we investigated how ATM responds to ROSs in Caenorhabditis elegans (C. elegans). Materials and Methods. First, we measured sensitivities of larvae to DNA-damaging agents and ROSs. Next, we analyzed the drug sensitivities of fully matured adult worms, which consist of nondividing somatic cells. Dead cell staining with acridine orange was performed to visualize the dead cells. In addition, we performed GFP reporter assays of lgg-1, an autophagy-related gene, to determine the types of cell death. Results. atm-1(tm5027) larvae showed a wide range of sensitivities to both DNA-damaging agents and ROSs. In contrast, fully matured adult worms, which consist of nondividing somatic cells, showed sensitivity to DNA-damaging agent, NaHSO3, but they showed resistance to H2O2. Dead cell staining and GFP reporter assays of lgg-1 suggest that C. elegans ATM-1 induces the cell death with autophagy in intestinal cells in response to H2O2. Conclusion. We revealed that ATM induces cell death in response to H2O2.

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 mRNA localization is linked to translation regulation in the Caenorhabditis elegans germ lineage

2020 ◽  
Author(s):  
Dylan M. Parker ◽  
Lindsay P. Winkenbach ◽  
Samuel P. Boyson ◽  
Matthew N. Saxton ◽  
Camryn Daidone ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Regulatory Control ◽  
Translational Repression ◽  
Translation Regulation ◽  
Germline Development ◽  
P Granules ◽  
C Elegans ◽  
Early Embryos

AbstractCaenorhabditis elegans early embryos generate cell-specific transcriptomes despite lacking active transcription. This presents an opportunity to study mechanisms of post-transcriptional regulatory control. In seeking the mechanisms behind this patterning, we discovered that some cell-specific mRNAs accumulate non-homogenously within cells, localizing to membranes, P granules (associated with progenitor germ cells in the P lineage), and P-bodies (associated with RNA processing). Transcripts differed in their dependence on 3’UTRs and RNA Binding Proteins, suggesting diverse regulatory mechanisms. Notably, we found strong but imperfect correlations between low translational status and P granule localization within the progenitor germ lineage. By uncoupling these, we untangled a long-standing question: Are mRNAs directed to P granules for translational repression or do they accumulate there as a downstream step? We found translational repression preceded P granule localization and could occur independent of it. Further, disruption of translation was sufficient to send homogenously distributed mRNAs to P granules. Overall, we show transcripts important for germline development are directed to P granules by translational repression, and this, in turn, directs their accumulation in the progenitor germ lineage where their repression can ultimately be relieved.SummaryMaternally loaded mRNAs localize non-homogeneously within C. elegans early embryos correlating with their translational status and lineage-specific fates.

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.

Segregation of germ granules in living Caenorhabditis elegans embryos: cell-type-specific mechanisms for cytoplasmic localisation

Development ◽  
1996 ◽  
Vol 122 (4) ◽  
pp. 1303-1312 ◽  
Author(s):  
S.N. Hird ◽  
J.E. Paulsen ◽  
S. Strome
Keyword(s):  
Caenorhabditis Elegans ◽  
Laser Scanning ◽  
Daughter Cell ◽  
Laser Scanning Confocal Microscopy ◽  
P Granules ◽  
C Elegans ◽  
Scanning Confocal Microscopy ◽  
Germ Granules ◽  
Cell Type Specific ◽  
P Cells

Germ granules are ribonucleoprotein particles that are thought to function in germline specification in invertebrates and possibly in vertebrates. In Caenorhabditis elegans, these structures, termed P granules, are partitioned to the germline P cells during the early embryonic divisions. By injecting a fluorescently labelled anti-P-granule antibody into the C. elegans germline syncitium, we followed P-granule segregation in live embryos using laser-scanning confocal microscopy. We show that, in early P cells (P0 and P1), P-granule partitioning is achieved primarily by their migration through the cytoplasm towards the site of formation of the germline daughter cell. A different mechanism appears to operate in later P cells (P2 and P3): P granules associate with the nucleus and move with it toward the site of formation of the germline daughter cell, where they are then deposited. At each division, there is also disassembly or degradation of those P granules that remain in the cytoplasm destined for the somatic daughter cell. Microfilaments, microtubules and the product of the gene mes-1 are required for the normal pattern of P-granule segregation in P2.

scholarly journals The isolation and in situ location of adligin: the microtubule cross-linking protein from Caenorhabditis elegans.

1989 ◽  
Vol 108 (3) ◽  
pp. 955-963 ◽  
Author(s):  
E Aamodt ◽  
R Holmgren ◽  
J Culotti
Keyword(s):  
Caenorhabditis Elegans ◽  
Structural Protein ◽  
Microtubule Network ◽  
C Elegans ◽  
Exclusion Chromatography ◽  
Associated Proteins ◽  
Cross Links ◽  
The Cross ◽  
Labeling Pattern

Microtubules isolated from the nematode Caenorhabditis elegans contain long stretches of periodic cross-links formed by microtubule-associated proteins (MAPs). These cross-links are 5.7 nm long, 3 nm wide, and occur at one tubulin dimer (8-nm) intervals along the walls of microtubules (Aamodt, E., and J. Culotti, 1986. J. Cell Biol. 103:23-31). The structural protein of the cross-links was isolated from the MAPs by centrifugation and exclusion chromatography. The cross-links were formed exclusively from the most prevalent MAP, a 32,000 mol wt protein. We suggest the name adligin for this MAP. Adligin eluted from the exclusion column at 33,000 mol wt indicating that it was a monomer in solution. Antibodies were made against the purified adligin and affinity purified. The affinity-purified antibodies were used to locate adligin in situ and to determine its distribution relative to that of tubulin by the use of double label immunofluorescence. The anti-adligin antibodies labeled a fibrous network in the cytoplasm of most cells of C. elegans. Neurons were labeled especially well. This labeling pattern was similar to the labeling pattern obtained with antitubulin, but anti-adligin labeled some granules in the gut that were not labeled with antitubulin. These results suggest that adligin may be part of the interphase microtubule network in C. elegans.

scholarly journals Caenorhabditis elegans β-G Spectrin Is Dispensable for Establishment of Epithelial Polarity, but Essential for Muscular and Neuronal Function

2000 ◽  
Vol 149 (4) ◽  
pp. 915-930 ◽  
Author(s):  
Suraj Moorthy ◽  
Lihsia Chen ◽  
Vann Bennett
Keyword(s):  
Nervous System ◽  
Caenorhabditis Elegans ◽  
Plasma Membranes ◽  
Striated Muscle ◽  
Expression Patterns ◽  
Cell Contact ◽  
Epithelial Polarity ◽  
Cell Stage ◽  
C Elegans ◽  
Cell Cell

The Caenorhabditis elegans genome encodes one α spectrin subunit, a β spectrin subunit (β-G), and a β-H spectrin subunit. Our experiments show that the phenotype resulting from the loss of the C. elegans α spectrin is reproduced by tandem depletion of both β-G and β-H spectrins. We propose that α spectrin combines with the β-G and β-H subunits to form α/β-G and α/β-H heteromers that perform the entire repertoire of spectrin function in the nematode. The expression patterns of nematode β-G spectrin and vertebrate β spectrins exhibit three striking parallels including: (1) β spectrins are associated with the sites of cell–cell contact in epithelial tissues; (2) the highest levels of β-G spectrin occur in the nervous system; and (3) β spec-trin-G in striated muscle is associated with points of attachment of the myofilament apparatus to adjacent cells. Nematode β-G spectrin associates with plasma membranes at sites of cell–cell contact, beginning at the two-cell stage, and with a dramatic increase in intensity after gastrulation when most cell proliferation has been completed. Strikingly, depletion of nematode β-G spectrin by RNA-mediated interference to undetectable levels does not affect the establishment of structural and functional polarity in epidermis and intestine. Contrary to recent speculation, β-G spectrin is not associated with internal membranes and depletion of β-G spectrin was not associated with any detectable defects in secretion. Instead β-G spectrin-deficient nematodes arrest as early larvae with progressive defects in the musculature and nervous system. Therefore, C. elegans β-G spectrin is required for normal muscle and neuron function, but is dispensable for embryonic elongation and establishment of early epithelial polarity. We hypothesize that heteromeric spectrin evolved in metazoans in response to the needs of cells in the context of mechanically integrated tissues that can withstand the rigors imposed by an active organism.

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