Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Epithelial structure revealed by chemical dissection and unembedded electron microscopy.

E G Fey, D G Capco, G Krochmalnic

    The Journal of Cell Biology
    |July 1, 1984
    PubMed
    Summary

    This study used chemical dissection and unembedded electron microscopy to examine cytoskeletal structures in epithelial cells. Researchers extracted Madin-Darby canine kidney cells with Triton X-100 and (NH4)2SO4 to reveal cytoskeletal networks. They found that the cytoskeleton includes a plasma lamina, filament networks, and desmosomes. Sequential extractions showed distinct structural layers with unique protein compositions. The remaining scaffold retained epithelial morphology and desmosomal junctions. The study suggests a structural continuum linking nuclear matrices, intermediate filaments, and desmosomes. The findings indicate that cytoskeletal fractions are biochemically distinct.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Morphological studies of cells grown in the absence of mitochondrial-specific protein synthesis.

    The Journal of cell biology·2009
    Same author

    Methylation and processing of ribosomal RNA in HeLa cells.

    Journal of molecular biology·2009
    Same author

    Human fibroblast growth factor 20 (FGF-20; CG53135-05): a novel cytoprotectant with radioprotective potential.

    International journal of radiation biology·2005
    Same author

    Spatial-temporal imaging of bacterial infection and antibiotic response in intact animals.

    Proceedings of the National Academy of Sciences of the United States of America·2001
    Same author

    Molecular and biochemical regulation of early mammalian development.

    International review of cytology·2001
    Same author

    Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors.

    Proceedings of the National Academy of Sciences of the United States of America·2001

    Area of Science:

    • Cellular biology
    • Electron microscopy techniques
    • Cytoskeletal structure analysis

    Background:

    Prior research has shown that cytoskeletal structures in epithelial cells form a complex network. Established knowledge includes the role of intermediate filaments and desmosomes in maintaining cell shape. However, the detailed structural relationships between cytoskeletal components and nuclear matrices remain unclear. This gap motivated the use of chemical dissection to explore cytoskeletal architecture. No prior work had resolved the biochemical uniqueness of cytoskeletal fractions. Researchers sought to determine if sequential extractions could reveal distinct structural layers. This paper's contribution is the first use of unembedded electron microscopy to examine cytoskeletal scaffolds. The study aimed to clarify how cytoskeletal networks retain epithelial morphology.

    Purpose Of The Study:

    The aim of this study was to examine cytoskeletal structures in epithelial cells using chemical dissection. The researchers focused on understanding how cytoskeletal networks retain epithelial morphology. They sought to determine if sequential extractions could reveal distinct structural layers. The motivation was to explore the structural continuum between nuclear matrices and desmosomes. The study aimed to clarify how cytoskeletal networks retain epithelial morphology. The researchers wanted to assess the biochemical uniqueness of cytoskeletal fractions. They also aimed to compare cytoskeletal structures in embedded and unembedded samples. The study sought to identify how cytoskeletal scaffolds maintain epithelial organization.

    Keywords:
    cytoskeletal structureelectron microscopyepithelial cell organizationcellular scaffold

    Frequently Asked Questions

    The cytoskeleton included a peripheral plasma lamina, filament networks, and desmosomes.

    It left structures enriched in intermediate filaments and desmosomes around nuclei.

    To remove chromatin and isolate the nuclear matrix-intermediate filament scaffold.

    It suggests a link between nuclear matrices, intermediate filaments, and desmosomes.

    Both showed retention of detailed morphological aspects of intact cells.

    Related Experiment Videos

    Main Methods:

    The study used Madin-Darby canine kidney epithelial cell monolayers for analysis. Cells were extracted with Triton X-100 in a nearly physiological buffer. Transmission electron micrographs were taken of whole mounts and thick sections. Sequential extractions with (NH4)2SO4 were used to fractionate cytoskeletal components. Nuclease digestion and (NH4)2SO4 elution were applied to remove chromatin. The remaining structures were examined in both embedded and unembedded samples. Researchers compared cytoskeletal networks in different extraction conditions. Protein composition was analyzed in four distinct cytoskeletal fractions.

    Main Results:

    Extraction with Triton X-100 revealed a cytoskeleton with plasma lamina and filament networks. (NH4)2SO4 extraction left structures enriched in intermediate filaments and desmosomes. Nuclease digestion removed chromatin, leaving a stable scaffold with cytokeratin filaments. The remaining structure retained epithelial morphology and desmosomal junctions. Cytoskeletal networks showed structural continuity with nuclear matrices and desmosomes. Protein composition of each fraction was found to be biochemically distinct. The cytoskeleton contained a peripheral plasma lamina and filament networks. The nuclear matrix-intermediate filament scaffold remained after chromatin removal.

    Conclusions:

    The study suggests that cytoskeletal structures form a scaffold for epithelial organization. The cytoskeleton contains a plasma lamina, filament networks, and desmosomes. Sequential extractions revealed distinct structural layers with unique protein compositions. The stable scaffold retained epithelial morphology and desmosomal junctions. The nuclear matrix-intermediate filament scaffold links to cytoskeletal networks. Protein composition differences were observed in each cytoskeletal fraction. The structural continuum between nuclear matrices and desmosomes was maintained. The findings suggest a biochemical distinction among cytoskeletal components.

    Each fraction had a unique protein composition, suggesting biochemical distinctions.