WormBase Tree Display for Expr_pattern: Expr2256
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| Expr2256 | Expression_of | Gene | WBGene00002368 | |
|---|---|---|---|---|
| Reflects_endogenous_expression_of | WBGene00002368 | |||
| Expression_data | Life_stage | WBls:0000003 | ||
| Anatomy_term | WBbt:0007028 | Certain | ||
| GO_term | GO:0005818 | |||
| GO:0005737 | ||||
| GO:0051233 | ||||
| Subcellular_localization | At the two-cell stage, LET-99 was enriched in a band at the cell periphery of P1 at all stages of the cell cycle (n=47). While the cell-cell contact region also appeared enriched for LET-99, quantification revealed this region had a staining intensity consistent with the juxtaposition of the AB and P1 membranes. Thus, LET-99 appears to be present in three domains at the cell periphery in P1 just as in the one-cell embryo. During third cleavage, LET-99 was enriched in a peripheral band in P2, and the band was in a position consistent with the reversal of polarity exhibited by this cel. In contrast to the P lineage cells, the AB cells and EMS had symmetric distributions of LET-99 at all times (n=43). Peripheral LET-99 became weaker in later embryos, disappearing between the 28-cell and the 50-cell stage. | |||
| In mitotic-stage wild-type embryos, LET-99 was asymmetrically enriched at the cell periphery, beginning at pronuclear migration in the one-cell embryo. The areas enriched for LET-99 encircled the posterior of the embryo but did not include the entire pole, and will be referred as posterior bands; one-cell embryos thus exhibited three distinct regions of LET-99 staining: the anterior domain, the posterior band and the posterior domain. The posterior bands were asymmetrically positioned at all stages; for example, in embryos at nuclear rotation stage, the posterior band extended from 51-74% egg length. Quantification of average fluorescence intensity confirmed that the highest staining intensity was in the posterior band at all stages and that the intensity increased during the cell cycle. In late anaphase embryos, a strong LET-99 band was still present, but the staining intensity of the posterior domain diminished. | ||||
| LET-99 was present in the cytoplasm and asymmetrically enriched at the cell periphery during both meiosis and mitosis. All of the patterns described are specific to LET-99, as evidenced by their absence in let-99 mutant embryos. In wild type, LET-99 was first observed during meiosis I and II where it was asymmetrically localized to the anterior periphery near meiotic spindles (37/44 embryos); both polar bodies were positive for LET-99 in later embryos. In embryos in which meiosis occurred laterally or at the posterior pole, LET-99 was always associated with the periphery adjacent to the meiotic spindle (n=9). Thus, LET-99 is localized near the meiotic spindle and not to the anterior of the embryo per se. In addition, LET-99 was observed between opposed chromatin masses in metaphase and in the spindle midzone at anaphase during meiosis and mitosis. During mitosis, a slight enrichment of LET-99 in the cytoplasm around nuclei and microtubule asters was also seen. However, no gross defects in mitosis, meiosis or polar body formation were observed in let-99 mutants or let-99(RNAi) embryos. | ||||
| Type | Antibody | Polyclonal rabbit antibody. A fragment of a let-99 cDNA, corresponding to amino acids 168-462, was cloned into the pMAL protein purification vector, expressed in bacteria, purified using amylose resin and injected into rabbits. Antisera were purified using a GST:LET-99 fusion protein (pGEX) coupled to affigel. In situ immunolocalization experiments was carried out using affinity purified antibodies specific for the LET-99 protein to determine the localization of LET-99 in embryos. | ||
| Pattern | Expressed in embryos. Expression pattern in other life stages was not analyzed. | |||
| Picture | WBPicture0000007517 | |||
| WBPicture0000007518 | ||||
| Reference | WBPaper00005515 | |||
| Antibody_info | WBAntibody00000564 |
