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Expr14682
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We first show the localization of UNC-59 at the cleavage furrow (previously shown with antibody staining, (Nguyen et al. 2000)) during a time lapse of cell divisions in early 2- to 4-cell stages of embryogenesis and throughout embryogenesis (cleavage rings in older embryos. Septins are also important for gonad morphogenesis and distal tip cell (DTC) migration (Nguyen et al. 2000) where UNC-59 protein is detected throughout gonad development in the rachis (previously shown with endogenously tagged unc-59::mKate, (Priti et al. 2018)) and DTCs. We highlight UNC-59/Septin localization in the DTC (previously shown with a transgene, (Finger et al. 2003)) at the L2 and L3 stages where it is organized into bundles (DeMay et al. 2011) and ring structures. The two bilateral sex myoblast cells express UNC-59 during their posterior to anterior migration in the L2 and early L3 stage and continue to express UNC-59 in these cells as they differentiate into vulval muscles in the late L3 to early L4 stages. Lastly, we show UNC-59/Septin expression and localization in tissue not previously reported: in the pharynx (cells of the buccal cavity, anterior procorpus, and terminal bulb); in the seam cells, both in bundles and at the cleavage furrows, beginning in the L1 stage and continuing throughout development and into the adult; and in sperm surrounding an embryo that has exited the spermatheca. |
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Expr12265
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The localization of tetraThymosin-beta is shown in oocytes (in dissected gonads of adult worms), in embryos and in whole adult organisms. TetraThymosin-beta was maternally expressed and in the adult gonad diffusely present in the cytoplasm of the oocytes, with strong staining at the cell cortex where also actin is present. In the distal arm of the gonad, tetraThymosin-beta localized to the inside edges of the membrane cubicles surrounding germ cell nuclei. Here, tetraThymosin-beta did not localize to the lateral side of the membrane cubicles where actin is arranged in a 'honeycomb' structure (Strome, 1986). In two-cell stage embryos, tetraThymosin-beta was enriched in the cell-cell contact where actin was also concentrated. The tetraThymosin-beta signal was clearly apparent until the four-cell stage and became weaker later on. At the comma stage (290 min after the first cell division), tetraThymosin-beta staining was observed at the developing nerve ring that is the largest axonal bundle in the nematode body and that positions around the isthmus of the pharynx. TetraThymosin-beta was evident in this region even before a clear actin signal was apparent. In larvae and adults, immunofluorescence yielded a diffuse but specific staining of the entire worm body with enrichment at the intestinal tract and spermatheca. At these stages, in contrast to what is observed in developing embryo, no prominent signal was observed in the head region where the nerve ring is located. In young adults is expressed in theposterior bulb of the pharynx. These immunostaining patterns were reproducible in three or more experiments and in 20 or more worms or embryos. |
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Expr12428
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NEG-1::GFP is asymmetrically localized in the early embryo. NEG-1::GFP was detected in the zygotic nucleus and at equal levels in both nuclei of the two-cell embryo (23 of 23). However, at the four-cell stage, NEG-1::GFP expression was markedly higher in nuclei of the anterior AB blastomeres than in the nuclei of EMS and P2 (31 of 34). Following the four-cell stage, NEG-1::GFP remained high in the granddaughters of the AB blastomere and diminished progressively in subsequent divisions (data not shown). In the adult germline, NEG- 1::GFP was observed in the nuclei of distal germ cells and the nuclei of growing oocytes except for the most proximal oocyte. Moreover, intense sub-nuclear localization of NEG-1:GFP was observed on condensed chromatin. |
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Expr12331
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Wild-type animals showed ubiquitous and uniform expression restricted to the cytoplasm as early as the two cell stage. This pattern persisted to the onset of elongation. RHGF-2 protein expression may be stronger in the seam cells post-elongation, around the 3-fold stage. The RHGF-2 antibody also detected expression in the nervous system starting around the 1.5 fold stage. RHGF-2 also lined the borders of cell in the pharyngeal lumen. |
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Subcellular localization: GLP-1 was found in the cytoplasm at the 2-cell stage, then in cytoplasm and membranes at 4-cell and 8-cell stages. Cytoplasmic GLP-1 fades after the 8-cell stage, and disappears by the 28-cell stage. Membrane-associated GLP-1 is faint by the 28-cell stage. The glp-1 mRNA was distributed uniformly through the 8-cell stage. Levels of glp-1 mRNA decline after the 8-cell stage and largely disappear by the 28-cell stage, though signal consistently persisted later in posterior parts of embryos. mRNA reappears after 100-cell stage, paralleling immunostaining results. early embryo(author) = blastula + gastrulating embryo(curator). |
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Expr541
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Faint expression in AB at 2-cell stage, becoming stronger in AB descendants after 4-cell stage. Signal weakens, between the 8- and 28-cell stages. GLP-1 not detected again until after the 100-cell stage in unidentified cells. |
GLP-1 was found in the cytoplasm at the 2-cell stage, then in cytoplasm and membranes at 4-cell and 8-cell stages. Cytoplasmic GLP-1 fades after the 8-cell stage, and disappears by the 28-cell stage. Membrane-associated GLP-1 is faint by the 28-cell stage. The glp-1 mRNA was distributed uniformly through the 8-cell stage. Levels of glp-1 mRNA decline after the 8-cell stage and largely disappear by the 28-cell stage, though signal consistently persisted later in posterior parts of embryos. mRNA reappears after 100-cell stage, paralleling immunostaining results. |
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Expr2582
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The OMA-1::GFP fluorescence in the developing oocytes and one-cell embryos recapitulated the wild-type spatial and temporal patterns of OMA-1 antibody staining. The punctate staining appeared more pronounced and resembled the characteristic pattern of germline P granules. Starting with the onset of the first mitotic division, the intensity of OMA-1-GFP fluorescence rapidly decreased, and by the time the division was complete, only approximately 10% remained. Interestingly, that remaining 10% of the GFP signal in the two-cell embryo was predominantly found in the germline precursor, P1, associated with what appeared to be P granules. The GFP signal continued to decrease in two-cell embryos and again was asymmetric after the next division, with most of the remaining fluorescence segregated to P2, where it was also predominantly associated with granules. The OMA-1-GFP signal became too weak to detect in the embryo after the four-cell stage. |
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Expr10817
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GFP:MEX-6 signal was enriched in the anterior blastomere of the two-cell embryo and in this blastomere's daughters at the early four-cell stage. |
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This information was extracted from published material (Archana Sharma-Oates, Andrew Mounsey and Ian A. Hope). |
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Expr746
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pal-1 RNA evenly distributed in 1- and 2-cell embryos. In 4-cell embryos, pal-1 RNA was either uniformly distributed (approximately 50%) or preferentially localized to the posterior blastomere. Nuclear localized transcripts were not detected before the 24-cell stage. In 6 cell and older embryo, pal-1 RNA was more abundant in cells that express PAL-1 protein. |
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early embryo(author) = 1-cell + 2-cell + 4-cell embryo(curator). |
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Expr573
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mex-3 mRNA is detected in the syncytial core of gonad arm and distribute uniformly in oocyte and early 1-cell embryo. mex-3 mRNA is more abundant in AB and its daughter cells than P1 and its daughter cells. After 4-cell stage, mex-3 mRNA disappears. MEX-3 protein is present in oocytes and uniformly distributed until the first division, when they become more abundant in the anterior AB cell. At the 4-cell stage, AB daughters have more MEX-3. Both the mRNA and protein disappear after the 4-cell stage. |
MEX-3 is cytoplasmic, and in the P cells, MEX-3 is associated with P granules. |
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Expr14312
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GFP tagged Myosin and Anillin, expressed under endogenous promoters, are visible in the cortex and contractile ring in 1 to 4 cell stage embryos. |
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Expr9127
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Expressed in the cytoplasm with increasing levels in growing oocytes, peaking in the oocyte undergoing maturation. The level of OMA-1 protein remain very high following fertilization as well as throughout the first mitotic cycle. The antibody staining often included a slight punctate pattern of fluorescence. Immediately after the first mitotic division, the levels of OMA-1 protein rapidly decreased and became difficult to detect with antibody staining. |
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Expr2581
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Expressed in the cytoplasm with increasing levels in growing oocytes, peaking in the oocyte undergoing maturation. The level of OMA-2 protein remain very high following fertilization as well as throughout the first mitotic cycle. The antibody staining often included a slight punctate pattern of fluorescence. Immediately after the first mitotic division, the level of OMA-2 proteins rapidly decreased and became difficult to detect with antibody staining. |
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No detailed cellular pattern description later than 2 cell stage embryo. |
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Expr1035
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Overall, OOC-3 is localized to a membrane compartment that had a dynamic distribution through the cell cycle. From fertilization until the beginning of pronuclear migration OOC-3 is uniformly enriched at the cortex of P0 in 75% of the embryos (n=36). At the beginning of pronuclear migration the cortical staining diminishes and OOC-3 is enriched around pronuclei (n=10). During rotation in P0, OOC-3 redistributes to the asters and in mitosis OOC-3 becomes enriched at the mitotic spindle. Towards the beginning of prophase of two-cell stage embryos OOC-3 localizes to a reticular structure spanning the whole cytoplasm, being enriched at the circumference of nuclei. In mitosis it again localizes to the mitotic spindle. The reticular localization can be more clearly seen in a 3-fold enlargement taken using a deconvolution microscope. ER localization was confirmed by staining C. elegans embryos with an antibody to HDEL, a sequence responsible for retention of proteins in the ER. OOC-3 is slightly enriched at the cell-cell boundary between AB and P1 in 50% of the embryos examined (n=45). |
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No detailed description on cellular expression pattern at later stages. |
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Expr1558
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No staining was seen in gonads or unfertilized oocytes. The PAR-3 protein was first detected at the cell periphery in fertilized eggs during the meiotic divisions. In 10 out of 13 positively stained embryos, the protein was undetectable in the posterior third of the embryo. stained very strongly on the lateral periphery, and stained weakly at the periphery at the extreme anterior pole. The other embryos showed uniform peripheral staining. After meiosis II is completed and as the pronuclei start to decondense, PAR-3 is restricted to the anterior periphery at all cases, extending to about 65% of the total length of the embryo. As pronuclear migration processes, PAR-3 become more concentrated at the anterior pole, extending to about 54% of the embryonic length. During prophase, metaphase, and early anaphase of the first mitosis, the signal is strongest and extends to 48% embryo length. In addition to peripheral staining, at all stages, unlocalized PAR-3 appears to be present in the cytoplasm. |
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No detailed description on cellular expression pattern of later developmental stages. |
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Expr1254
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Staining of DNC-1 in one-cell and two-cell embryos revealed a striking accumulation at the cell-division remnant (also known as the residual body or midbody). |
Centrosomal and kinetochore staining was also seen. In addition, cytoplasmic staining was seen. Initially, the accumulation of DNC-1 at cell-division remnants appeared upon completion of the first meiosis during polar-body formation. This spot persisted until the end of the first mitosis. DNC-1 appeared at the leading edge of the cleavage furrow of one-cell stage embryos, and persisted at the cell-division remnant until the end of the two-cell stage. |
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Deconvolution microscopy to generate high resolution images of the nuclei of two-cell embryos stained with mAb 6C4 and DAPI. The relative time of each nucleus in the cell cycle was inferred by comparing the staining patterns of the AB and P1 blastomeres; the AB blastomere division occurs before that of the P1 blastomere. The morphology of DAPI staining and presence of a nuclear membrane were used as additional landmarks of cell cycle progression. No other cellular expression pattern described. |
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Expr1191
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Interphase nuclei had no detectable mAb 6C4 reactivity. Nuclei that are in the earliest stages of chromosome condensation do not stain with mAb 6C4. Staining with mAb 6C4 was first observed in prophase nuclei that contain condensed chromosomes. At this stage, mAb 6C4 staining was observed as dots distributed throughout the nucleus; many of these dots colocalize with the chromosomes. These chromosome-associated dots are widely dispersed along the entire length of each chromosome. Later in prophase, mAb 6C4 stained structures located on opposite sides of each chromosome. In metaphase, when the chromosomes are aligned at the equatorial plane, chromosomes were oriented with each mAb 6C4 stained structure facing one centrosome. At anaphase, sister chromatids separated with the mAb 6C4 stained side on the poleward face of each sister chromatid. No mAb 6C4 staining was detectable by late anaphase. These results show that there is dynamic localization of the mAb 6C4 antigen to chromosomes throughout mitosis. mAb 6C4 Staining during meiosis During meiosis, C. elegans chromosomes lack a trilaminar kinetochore structure, and attachment to the meiotic spindle is mediated through the chromosome ends. No mAb 6C4 nuclear staining was detected in the mitotic region of the gonad of the hermaphrodite nor in oocytes arrested at diakinesis of prophase I. After fertilization, embryos exit prophase I arrest and undergo two consecutive rounds of division to form two polar bodies. Staining with mAb 6C4 was detected on meiotic chromosomes after fertilization and before first olar body formation. The mAb 6C4 staining pattern was observed as a halo surrounding the meiotic chromosomes. |
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Expr13177
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Early embryos were fixed and stained with Mab F2F4 (green), shown to recognize CYB-3 (Michael, 2016), and DAPI, to illuminate the DNA (blue). Either wild type or par-4 mutant embryos were examined, after 24-hour incubation at 25C (the non-permissive temperature for the it47 allele of par-4). Anterior is to the left in all images. The data presented here reveals previously not shown data that depicts CYB-3 as asymmetrically distributed at the 4-cell stage. These data further support reported findings in Michael, 2016. There is more CYB-3 in the AB cell relative to its sister P1. In 4-cell embryos there is more CYB-3 in the EMS cell relative to its sister, P2. Thus, during P-lineage divisions, CYB-3 is asymmetrically distributed such that the somatic precursor receives more than its germline precursor sister cell. This asymmetry is abolished in par-4 mutant embryos, where all blastomeres contain equivalent amounts of CYB-3. |
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Expr13457
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DHC-1::GFP was readily detectable on growing MT plus ends in early embryos. dhc-1::gfp knock-in allele. |
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Expr13024
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ATX-2: exhibits diffuse cytoplasmic expression in early embryo with occasional punctate foci. |
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Expr14311
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GFP tagged Myosin and Anillin, expressed under endogenous promoters, are visible in the cortex and contractile ring in 1 to 4 cell stage embryos. |
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No detailed description on cellular localization at later stages. |
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Expr1287
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Blastula embryo. |
The antibodies stained the cell periphery of early blastomeres as well as the nuclear envelope. Nuclear envelope staining however, appears to be non-specific. The staining in 1-cell embryos is weak and uniform just after the completion of meiosis, but increases in intensity and becomes concentrated at the anterior periphery during pronuclear migration. The peripheral PKC-3 staining becomes restricted to about 50% embryo length during the pronuclear meeting and pronuclear fusion stage (n=23). By early anaphase, the staining extends posteriorly beyond the midline of the zygote and covers about 60% of the total length of embryos (n=29). In 2- and 4-cell stages, staining is uniform at the periphery of the AB cell, its daughters and the EMS cell, but peripheral staining in P1 and P2 is restricted to the boundaries with other blastomeres. |
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To confirm that the localization patterns described are specific to POD-1, pod-1(ye11) mutant embryos were stained with both POD-1 antibodies. All cortical and cytoplasmic POD-1 staining was eliminated in the mutant embryos. |
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Expr1162
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POD-1 localization in P cells is cell cycle dependent. In the earliest one-cell embryos before pronuclear meeting (P0), cortical POD-1 is either anteriorly enriched (12/24 embryos), posteriorly enriched (5/24 embryos), or uniformly localized (7/24 embryos). From pronuclear meeting to metaphase, cortical POD-1 is almost always anteriorly enriched (30/39 embryos). At anaphase, POD-1 begins to transition from asymmetric to symmetric localization, such that by telophase cortical POD-1 can be found at equivalent levels in the anterior and posterior cortices (10/11 embryos). In addition to cell cortices, POD-1 antibodies stain punctate cytoplasmic structures. POD-1 can be found in the posterior-most cortex, albeit at lower levels than the anterior. The localization of POD-1 in the germ-line precursors of the two-cell and four-cell embryo is also cell cycle dependent such that early in the cell cycle, POD-1 is not asymmetric along the a-p axis, but becomes anteriorly enriched leading up to metaphase. |
punctate cytoplasmic structures |
