Contrasting patterns of mitochondrial redistribution in the early lineages of Caenorhabditis elegans and Acrobeloides sp. PS1146

Badrinath, Ananth; White, John

doi:10.1016/s0012-1606(03)00102-7

Cited by 13 publications

(10 citation statements)

References 22 publications

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“…We found that CAR-1-positive particles also labeled with the nucleic acid label PI ( Figure 4C), indicating that these small puncta, as well as the P-granules, contain RNA. This pattern of cytoplasmic PI labeling was clearly distinct from the published pattern of mitochondria organization (Badrinath and White, 2003).…”

Section: Large Car-1 Puncta Are P-granules and The Small Puncta Are Pcontrasting

confidence: 48%

CAR-1, a Protein That Localizes with the mRNA Decapping Component DCAP-1, Is Required for Cytokinesis and ER Organization inCaenorhabditis elegansEmbryos

Squirrell

Eggers

Luedke

et al. 2006

MBoC

102

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The division of one cell into two requires the coordination of multiple components. We describe a gene, car-1, whose product may provide a link between disparate cellular processes. Inhibition of car-1 expression in Caenorhabditis elegans embryos causes late cytokinesis failures: cleavage furrows ingress but subsequently regress and the spindle midzone fails to form, even though midzone components are present. The localized accumulation of membrane that normally develops at the apex of the cleavage furrow during the final phase of cytokinesis does not occur and organization of the endoplasmic reticulum is aberrant, indicative of a disruption in membrane trafficking. The car-1 gene has homologues in a number of species, including proteins that associate with RNA binding proteins. CAR-1 localizes to P-granules (germ-line specific ribonucleoprotein particles) and discrete, developmentally regulated cytoplasmic foci. These foci also contain DCAP-1, a protein involved in decapping mRNAs. Thus, CAR-1, a protein likely to be associated with RNA metabolism, plays an essential role in the late stage of cytokinesis, suggesting a novel link between RNA, membrane trafficking and cytokinesis in the C. elegans embryo.

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Section: Large Car-1 Puncta Are P-granules and The Small Puncta Are Pcontrasting

confidence: 48%

CAR-1, a Protein That Localizes with the mRNA Decapping Component DCAP-1, Is Required for Cytokinesis and ER Organization inCaenorhabditis elegansEmbryos

Squirrell

Eggers

Luedke

et al. 2006

MBoC

102

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“…In most organisms during interphase, mitochondria are localized throughout the cytoplasm. However, in some organisms such as yeast and the nematode Acrobeloides, mitochondria cluster around the centrosomes during mitosis, presumably to ensure proper segregation into separate daughter cells (Badrinath and White 2003;Yaffe et al 2003). This centrosome clustering is consistent with a possible dependence on dynein.…”

Section: Resultsmentioning

confidence: 64%

“…This centrosome clustering is consistent with a possible dependence on dynein. However, in C. elegans embryos, mitochondria do not undergo any obvious rearrangements during mitosis ( Figure 5) (Badrinath and White 2003), making SPD-3TGFP localization even more surprising. It is worth noting that mitochondrial morphology and localization as assessed with mitochondrial vital dyes were not affected in oj35 embryos, at least at levels that we could detect in these studies ( Figure 5).…”

Section: Resultsmentioning

confidence: 99%

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SPD-3 Is Required for Spindle Alignment in Caenorhabditis elegans Embryos and Localizes to Mitochondria

et al. 2007

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During the development of multicellular organisms, cellular diversity is often achieved through asymmetric cell divisions that produce two daughter cells having different developmental potentials. Prior to an asymmetric cell division, cellular components segregate to opposite ends of the cell defining an axis of polarity. The mitotic spindle rotationally aligns along this axis of polarity, thereby ensuring that the cleavage plane is positioned such that segregated components end up in individual daughter cells. Here we report our characterization of a novel gene required for spindle alignment in Caenorhabditis elegans. During the first mitosis in spd-3(oj35) embryos the spindle failed to align along the anterior/posterior axis, leading to abnormal cleavage configurations. spd-3(oj35) embryos had additional defects reminiscent of dynein/dynactin loss-of-function possibly caused by the mislocalization of dynactin. Surprisingly, we found that SPD-3TGFP localized to mitochondria. Consistent with this localization, spd-3(oj35) worms exhibited slow growth and increased ATP concentrations, which are phenotypes similar to those described for other mitochondrial mutants in C. elegans. To our knowledge, SPD-3 is the first example of a link between mitochondria and spindle alignment in C. elegans.

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“…This might include labeled small molecules, proteins, lipids, etc. In some cases, it may not be necessary to actually inject labeled trackers as the worms will readily uptake some dyes such as mitotracker (10). However, we have tried both soaking and injection of lysotracker and have found much better results with injection for visualization in embryos.…”

Section: Discussionmentioning

confidence: 99%

Time-lapse Microscopy of Early Embryogenesis in <em>Caenorhabditis elegans</em>

Boyd

Hajjar

O’Connell

2011

JoVE

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Caenorhabditis elegans has often been used as a model system in studies of early developmental processes. The transparency of the embryos, the genetic resources, and the relative ease of transformation are qualities that make C. elegans an excellent model for early embryogenesis. Laser-based confocal microscopy and fluorescently labeled tags allow researchers to follow specific cellular structures and proteins in the developing embryo. For example, one can follow specific organelles, such as lysosomes or mitochondria, using fluorescently labeled dyes. These dyes can be delivered to the early embryo by means of microinjection into the adult gonad. Also, the localization of specific proteins can be followed using fluorescent protein tags. Examples are presented here demonstrating the use of a fluorescent lysosomal dye as well as fluorescently tagged histone and ubiquitin proteins. The labeled histone is used to visualize the DNA and thus identify the stage of the cell cycle. GFP-tagged ubiquitin reveals the dynamics of ubiquitinated vesicles in the early embryo. Observations of labeled lysosomes and GFP:: ubiquitin can be used to determine if there is colocalization between ubiquitinated vesicles and lysosomes. A technique for the microinjection of the lysosomal dye is presented. Techniques for generating transgenenic strains are presented elsewhere (1, 2). For imaging, embryos are cut out of adult hermaphrodite nematodes and mounted onto 2% agarose pads followed by time-lapse microscopy on a standard laser scanning confocal microscope or a spinning disk confocal microscope. This methodology provides for the high resolution visualization of early embryogenesis. Video LinkThe video component of this article can be found at http://www.jove.com/video/2852/ Protocol 1. Nematode cultures 1. Obtain the appropriate C. elegans strain from the Caenorhabditis Genetics Stock Center (CGC) or from a colleague. 2. Grow nematodes on NGM agar plates seeded with an OP50 bacterial lawn (3). For analysis of GFP strains growth at 25°C is recommended. 3. The day before your microscopy experiment, pick at least 40 L4 larvae onto seeded plates and place the plates at 25°C overnight. These worms will be young adults for the experiment. InjectionsIf it is desirable to view structures such as lysosomes or mitochondria, adults can be injected with fluorescent dye prior to visualization.

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Contrasting patterns of mitochondrial redistribution in the early lineages of Caenorhabditis elegans and Acrobeloides sp. PS1146

Cited by 13 publications

References 22 publications

CAR-1, a Protein That Localizes with the mRNA Decapping Component DCAP-1, Is Required for Cytokinesis and ER Organization inCaenorhabditis elegansEmbryos

CAR-1, a Protein That Localizes with the mRNA Decapping Component DCAP-1, Is Required for Cytokinesis and ER Organization inCaenorhabditis elegansEmbryos

SPD-3 Is Required for Spindle Alignment in Caenorhabditis elegans Embryos and Localizes to Mitochondria

Time-lapse Microscopy of Early Embryogenesis in <em>Caenorhabditis elegans</em>

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