Epithelial structure revealed by chemical dissection and unembedded electron microscopy

E. G. Fey, David Capco, G. Krochmalnic, S. Penman

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Cytoskeletal structures obtained after extraction of Madin-Darby canine kidney epithelial cell monolayers with Triton X-100 were examined in transmission electron micrographs of cell whole mounts and unembedded thick sections. The cytoskeleton, an ordered structure consisting of a peripheral plasma lamina, a complex network of filaments, and chromatin-counseling nuclei, was revealed after extraction of intact cells with a nearly physiological buffer containing Triton X-100. The cytoskeleton was further fractionated by extraction with (NH4)2SO4, which left a structure enriched in intermediate filaments and desmosomes around the nuclei. A further digestion with nuclease and elution with (NH4)2SO4 removed the chromatin. The stable structure that remained after this procedure retained much of the epithelial morphology and contained essentially all of the cytokeratin filaments and desmosomes and the chromatin-depleted nuclear matrices. This structural network may serve as a scaffold for epithelial organization. The cytoskeleton and the underlying nuclear matrix-intermediate filament scaffold, when examined in both conventional embedded thin sections and in embedded whole mounts and thick sections showed that retention of many of the detailed morphological aspects of the intact cells, which suggests a structural continuum linking the nuclear matrix, the intermediate filament network, and the intercellular desmosomal junctions. Most importantly, the protein composition of each of the four fractions obtained by this sequential procedure was essentially unique. Thus the proteins constituting the soluble fraction, the cytoskeleton, the chromatin fraction, and the underlying nuclear matrix-intermediate filament scaffold are biochemically distinct.

Original languageEnglish (US)
JournalJournal of Cell Biology
Volume99
Issue number1 II
StatePublished - 1984
Externally publishedYes

Fingerprint

Nuclear Matrix
Intermediate Filaments
Cytoskeleton
Chromatin
Dissection
Electron Microscopy
Desmosomes
Octoxynol
Madin Darby Canine Kidney Cells
Intercellular Junctions
Keratins
Counseling
Digestion
Buffers
Proteins
Epithelial Cells
Electrons

ASJC Scopus subject areas

  • Cell Biology

Cite this

Epithelial structure revealed by chemical dissection and unembedded electron microscopy. / Fey, E. G.; Capco, David; Krochmalnic, G.; Penman, S.

In: Journal of Cell Biology, Vol. 99, No. 1 II, 1984.

Research output: Contribution to journalArticle

@article{db77283d699147e28cf922fa90c350d4,
title = "Epithelial structure revealed by chemical dissection and unembedded electron microscopy",
abstract = "Cytoskeletal structures obtained after extraction of Madin-Darby canine kidney epithelial cell monolayers with Triton X-100 were examined in transmission electron micrographs of cell whole mounts and unembedded thick sections. The cytoskeleton, an ordered structure consisting of a peripheral plasma lamina, a complex network of filaments, and chromatin-counseling nuclei, was revealed after extraction of intact cells with a nearly physiological buffer containing Triton X-100. The cytoskeleton was further fractionated by extraction with (NH4)2SO4, which left a structure enriched in intermediate filaments and desmosomes around the nuclei. A further digestion with nuclease and elution with (NH4)2SO4 removed the chromatin. The stable structure that remained after this procedure retained much of the epithelial morphology and contained essentially all of the cytokeratin filaments and desmosomes and the chromatin-depleted nuclear matrices. This structural network may serve as a scaffold for epithelial organization. The cytoskeleton and the underlying nuclear matrix-intermediate filament scaffold, when examined in both conventional embedded thin sections and in embedded whole mounts and thick sections showed that retention of many of the detailed morphological aspects of the intact cells, which suggests a structural continuum linking the nuclear matrix, the intermediate filament network, and the intercellular desmosomal junctions. Most importantly, the protein composition of each of the four fractions obtained by this sequential procedure was essentially unique. Thus the proteins constituting the soluble fraction, the cytoskeleton, the chromatin fraction, and the underlying nuclear matrix-intermediate filament scaffold are biochemically distinct.",
author = "Fey, {E. G.} and David Capco and G. Krochmalnic and S. Penman",
year = "1984",
language = "English (US)",
volume = "99",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "1 II",

}

TY - JOUR

T1 - Epithelial structure revealed by chemical dissection and unembedded electron microscopy

AU - Fey, E. G.

AU - Capco, David

AU - Krochmalnic, G.

AU - Penman, S.

PY - 1984

Y1 - 1984

N2 - Cytoskeletal structures obtained after extraction of Madin-Darby canine kidney epithelial cell monolayers with Triton X-100 were examined in transmission electron micrographs of cell whole mounts and unembedded thick sections. The cytoskeleton, an ordered structure consisting of a peripheral plasma lamina, a complex network of filaments, and chromatin-counseling nuclei, was revealed after extraction of intact cells with a nearly physiological buffer containing Triton X-100. The cytoskeleton was further fractionated by extraction with (NH4)2SO4, which left a structure enriched in intermediate filaments and desmosomes around the nuclei. A further digestion with nuclease and elution with (NH4)2SO4 removed the chromatin. The stable structure that remained after this procedure retained much of the epithelial morphology and contained essentially all of the cytokeratin filaments and desmosomes and the chromatin-depleted nuclear matrices. This structural network may serve as a scaffold for epithelial organization. The cytoskeleton and the underlying nuclear matrix-intermediate filament scaffold, when examined in both conventional embedded thin sections and in embedded whole mounts and thick sections showed that retention of many of the detailed morphological aspects of the intact cells, which suggests a structural continuum linking the nuclear matrix, the intermediate filament network, and the intercellular desmosomal junctions. Most importantly, the protein composition of each of the four fractions obtained by this sequential procedure was essentially unique. Thus the proteins constituting the soluble fraction, the cytoskeleton, the chromatin fraction, and the underlying nuclear matrix-intermediate filament scaffold are biochemically distinct.

AB - Cytoskeletal structures obtained after extraction of Madin-Darby canine kidney epithelial cell monolayers with Triton X-100 were examined in transmission electron micrographs of cell whole mounts and unembedded thick sections. The cytoskeleton, an ordered structure consisting of a peripheral plasma lamina, a complex network of filaments, and chromatin-counseling nuclei, was revealed after extraction of intact cells with a nearly physiological buffer containing Triton X-100. The cytoskeleton was further fractionated by extraction with (NH4)2SO4, which left a structure enriched in intermediate filaments and desmosomes around the nuclei. A further digestion with nuclease and elution with (NH4)2SO4 removed the chromatin. The stable structure that remained after this procedure retained much of the epithelial morphology and contained essentially all of the cytokeratin filaments and desmosomes and the chromatin-depleted nuclear matrices. This structural network may serve as a scaffold for epithelial organization. The cytoskeleton and the underlying nuclear matrix-intermediate filament scaffold, when examined in both conventional embedded thin sections and in embedded whole mounts and thick sections showed that retention of many of the detailed morphological aspects of the intact cells, which suggests a structural continuum linking the nuclear matrix, the intermediate filament network, and the intercellular desmosomal junctions. Most importantly, the protein composition of each of the four fractions obtained by this sequential procedure was essentially unique. Thus the proteins constituting the soluble fraction, the cytoskeleton, the chromatin fraction, and the underlying nuclear matrix-intermediate filament scaffold are biochemically distinct.

UR - http://www.scopus.com/inward/record.url?scp=0021219218&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021219218&partnerID=8YFLogxK

M3 - Article

C2 - 6540264

AN - SCOPUS:0021219218

VL - 99

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 1 II

ER -