dyf-7 Antibody
Description
Overview of DYF-7 Antibody
DYF-7 antibody is a specialized tool used to detect and study the DYF-7 protein, a zona pellucida (ZP) domain-containing extracellular matrix (ECM) protein critical for neuronal and epithelial morphogenesis in C. elegans. This antibody enables researchers to investigate DYF-7's roles in dendrite anchoring, apical ECM formation, and epithelial integrity through techniques such as immunofluorescence, Western blotting, and live imaging .
Localization Studies
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DYF-7 antibody detects DYF-7 at dendritic tips and epithelial apical surfaces. In C. elegans embryos, DYF-7 localizes to caps adjacent to tight junctions (marked by AJM-1) at dendrite endings, confirming its role in anchoring dendrites to epithelial structures .
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Misexpression experiments show DYF-7 ectodomain localization in gut and pharyngeal lumens, emphasizing its epithelial specificity .
Functional Analysis
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DYF-7 antibody identifies proteolytic cleavage events: DYF-7 undergoes furin-dependent cleavage, separating its ectodomain (tagged with sfGFP) from the membrane-bound cytoplasmic tail (mCherry-tagged) .
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Mutational studies reveal that deletion of the conserved furin cleavage site (CFCS) disrupts cleavage, leading to diffuse cytoplasmic localization .
Role in Dendrite Morphogenesis
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DYF-7 is required for dendrite extension in amphid, CEP, OL, and IL neurons. Mutants exhibit dendrite lengths reduced to ~10–80% of wild-type, with bimodal distributions indicating failed initiation events .
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DYF-7 collaborates with PAR-6 to ensure proper localization at rosette vertices. In par-6 mutants, DYF-7 becomes diffuse, leading to dendrite detachment .
Epithelial Integrity
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DYF-7 prevents rupture of glial pore tubes. Loss of DYF-7 disrupts sheath glial cell extension, causing lumen collapse .
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Redundant roles with SAX-7 and HMR-1 in dendrite-epidermis adhesion highlight its importance in collective cell migration .
Table 1: DYF-7 Mutations
Table 2: DYF-7 Constructs
| Construct | Tags/Modifications | Functionality | Localization |
|---|---|---|---|
| DYF-7ecto::sfGFP | Ectodomain GFP | Rescues dendrite defects | Dendrite caps, apical ECM |
| DYF-7cyt::mCherry | Cytoplasmic mCherry | Tracks membrane association | Diffuse neuronal membrane |
| HA-DYF-7∆CFCS-FLAG | HA/FLAG tags, CFCS deleted | Blocks cleavage | Cytoplasmic accumulation |
Technical Validation
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Specificity: DYF-7 antibody shows no cross-reactivity with other ZP domain proteins (e.g., FRM-2) .
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Sensitivity: Detects DYF-7 at nanomolar concentrations in reducing and non-reducing conditions (Fig. 6, ).
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Quantitative Use: Fluorescence-based detection systems enable linear signal quantification across orders of magnitude .
Implications for Human Disease
DYF-7 homologs in humans (e.g., uromodulin, tectorins) are linked to nephropathy, deafness, and cancer. Studying DYF-7’s role in apical ECM integrity provides insights into epithelial pathologies, including vascular dysplasia and Crohn’s disease .
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
