WormBase Tree Display for Expr_pattern: Expr2553
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| Expr2553 | Expression_of | Gene | WBGene00001069 | |
|---|---|---|---|---|
| Reflects_endogenous_expression_of | WBGene00001069 | |||
| Expression_data | Life_stage | WBls:0000024 | ||
| WBls:0000003 | ||||
| WBls:0000038 | ||||
| WBls:0000027 | ||||
| WBls:0000035 | ||||
| WBls:0000041 | ||||
| Anatomy_term | WBbt:0005733 | Certain | ||
| GO_term | GO:0032154 | |||
| GO:0031012 | ||||
| Subcellular_localization | The DPY-7 protein is detected in circumferential bands within the cuticle of each larval stage and the adult. The DPY-7 bands locate within the furrows that delineate the annuli. The DPY-7 bands are not continuous around the entire worm body. In the adult and the L1 larva, longitudinal ridges termed alae exist within the cuticle above the seam cells. DPY-7 was not detected within these ridges or the matrix region immediately surrounding them. In larval stages with no alae, the DPY-7 bands start and end above the lateral seam cells where they partly interdigitate. When a partial reduction step is included during fixation for immunostaining, the DPY-7containing bands dissociate from other ECM material and adopt a discrete thread-like appearance. The DPY-7 bands seem to be more resistant to reduction than the ECM material that lies between the bands. DPY-7 collagen probably assembled into tight band or thread-like structures that run circumferentially around the body of the animal, located within the furrows that delineate the annuli. | |||
| Type | Antibody | Mouse monoclonal antibody against DPY-7. Recombinant DPY-7 protein fragments were generated in Escherichia coli by using the QIAGEN vector pQE-30, according to standard manufacturer's protocol. A DPY-7 fragment constituting most of the predicted mature protein was used to immunize mice, and monoclonal cell lines were generated by fusing splenocytes to Sp2/O-Ag14 myeloma cells by using standard methods. The cell line DPY7-5a was selected for use based on the sensitivity and specificity of the antibody for the DPY-7 protein. It reacts to an epitope within the 40 carboxy-terminal non-Gly-X-Y residues of the protein. | ||
| Pattern | Intracellular localization of DPY-7 is first observed at the comma stage of embryogenesis, which corresponds to ~4 h before secretion of the L1 cuticle, concurring with the known temporal pattern of expression of dpy-7. The staining remains intracellular through the later stages of embryogenesis, and the first evidence of secretion of DPY-7 is at the threefold stage. Extracellular staining within the secreted cuticle is seen at the late threefold stage of embryogenesis just before hatch. DPY-7 location within the cells is detected as a halo surrounding the nucleus. | |||
| To assist in the identification of cells, the antibodies MH27 and anti-LIN-26 were used to visualize hypodermal cell junctions and nuclei of hypodermal cells, respectively. The DPY-7 protein is detected during embryogenesis in most and probably all hypodermal cells, and certainly within the hyp-7 cells that form the major body hypodermal syncytium, the P cells that form the ventral hypodermis, and the V cells that constitute the lateral seam. Although DPY-7 is synthesized in all major hypodermal cells, the mature secreted collagen is only detected on the apical surfaces of the dorsal and ventral hypodermal cells and not on the lateral surface above the seam cells of developmental stages that have alae. This absence was shown by costaining with the MH27 antibody to delineate the boundaries of the seam cells. | ||||
| Picture | WBPicture0000008719 | |||
| WBPicture0000008720 | ||||
| Reference | WBPaper00005815 | |||
| Antibody_info | WBAntibody00000615 |
