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The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging
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The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging

The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging

DOI:
10.3791/56101-v

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10:30 min

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October 03, 2017

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, , , , ,
1Mucosal Immunology and Biology Research Center, Developmental Biology and Genetics Core, Massachusetts General Hospital for Children,Harvard Medical School, 2College of Life Sciences,Jilin University, 3Faculty of Health Sciences,University of Macau

Chapters

  • 00:05Title
  • 01:02Introduction to the Accompanying Publication
  • 02:28RNAi Interaction Screen to Modify a Canal lumen Phenotype for Identification of Genes Involved in Intracellular Lumen Extension
  • 03:29Scoring Excretory Canal Phenotypes using Fluorescence Dissecting Microscopy
  • 05:20Imaging the C. elegans Excretory Canal using Confocal Microscopy
  • 08:16Results: GFP Labeling, RNAi and Microscopic Analysis of Excretory Canal Lumen Morphogenesis Phenotypes
  • 09:51Conclusion

Summary

자동 번역

자동 번역

The C. elegans excretory canal is a unique single-cell model for the visual in vivo analysis of de novo polarized membrane biogenesis. This protocol describes a combination of standard genetic/RNAi and imaging approaches, adaptable for the identification and characterization of molecules directing unicellular tubulogenesis, and apical membrane and lumen biogenesis.

Tags

C. ElegansExcretory CanalIntracellular LumenPolarized Membrane BiogenesisTubulogenesisPolarityGFP FusionRNAiLive ImagingMembrane MarkersOrganelle MarkersPromotersERM-1Cystic Canal PhenotypeModifier Screen

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