Cytoskeletal regulation of Caco‐2 intestinal monolayer paracellular permeability

Thomas Ma, Daniel Hollander, Le Thuy Tran, Don Nguyen, Neil Hoa, Deepak Bhalla

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

An abnormal increase in intestinal paracellular permeability may be an important pathogenic factor in various intestinal diseases. The intracellular factors and processes that regulate and cause alteration of intestinal paracellular permeability are not well understood. The purpose of this study was to examine some of the intracellular processes involved in cytoskeletal regulation of intestinal epithelial paracellular permeability using the filter‐grown Caco‐2 intestinal epithelial monolayers. Cytochalasin‐b and colchicine were used to disrupt the cytoskeletal elements, actin microfilaments, and microtubules. Cytochalasin‐b (5 m̈g/ml) and colchicine (2 × 10−5M) at the doses used caused marked depolymerization and disruption of actin microfilaments and microtubules, respectively. Cytochalasin‐b‐induced disruption of actin microfilaments resulted in perturbation of tight junctions and desmosomes and an increase in Caco‐2 monolayer paracellular permeability. The cytochalasin‐b‐induced disruption of actin microfilaments and subsequent changes in intercellular junctional complexes and paracellular permeability were not affected by inhibitors of protein synthesis (actinomycin‐D or cycloheximide) or microtubule function (colchicine), but were inhibited by metabolic energy inhibitors (2,4‐dinitrophenol or sodium azide). The cytochalasin‐b‐induced disturbance in Caco‐2 actin microfilaments and intercellular junctional complexes and increase in paracellular permeability were rapidly reversed. The paracellular pathway “re‐tightening” following cytochalasin‐b removal was not affected by actinomycin‐D, cycloheximide, or colchicine, but was inhibited by 2,4‐dinitrophenol and sodium azide. The colchicine‐induced disruption of microtubules did not have significant effect on actin microfilaments, intercellular junctions, or paracellular permeability. These findings suggest that cytochalasin‐b‐induced increase in Caco‐2 monolayer paracellular permeability was due to actin microfilament mediated perturbation of intercellular junctional complexes. The re‐tightening of paracellular pathways (following removal of cytochalasin‐b) resulted from energy‐mediated re‐assembly of pre‐existing actin microfilaments and intercellular junctional complexes. This re‐closure process did not require protein synthesis or microtubule‐mediated shuttling process. © 1995 Wiley‐Liss, Inc.

Original languageEnglish (US)
Pages (from-to)533-545
Number of pages13
JournalJournal of Cellular Physiology
Volume164
Issue number3
DOIs
StatePublished - Jan 1 1995

Fingerprint

Actin Cytoskeleton
Actins
Monolayers
Permeability
Colchicine
Microtubules
Sodium Azide
Cycloheximide
Intestinal Diseases
Desmosomes
Depolymerization
Protein Synthesis Inhibitors
Intercellular Junctions
Tight Junctions
Proteins

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Ma, Thomas ; Hollander, Daniel ; Tran, Le Thuy ; Nguyen, Don ; Hoa, Neil ; Bhalla, Deepak. / Cytoskeletal regulation of Caco‐2 intestinal monolayer paracellular permeability. In: Journal of Cellular Physiology. 1995 ; Vol. 164, No. 3. pp. 533-545.
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abstract = "An abnormal increase in intestinal paracellular permeability may be an important pathogenic factor in various intestinal diseases. The intracellular factors and processes that regulate and cause alteration of intestinal paracellular permeability are not well understood. The purpose of this study was to examine some of the intracellular processes involved in cytoskeletal regulation of intestinal epithelial paracellular permeability using the filter‐grown Caco‐2 intestinal epithelial monolayers. Cytochalasin‐b and colchicine were used to disrupt the cytoskeletal elements, actin microfilaments, and microtubules. Cytochalasin‐b (5 m̈g/ml) and colchicine (2 × 10−5M) at the doses used caused marked depolymerization and disruption of actin microfilaments and microtubules, respectively. Cytochalasin‐b‐induced disruption of actin microfilaments resulted in perturbation of tight junctions and desmosomes and an increase in Caco‐2 monolayer paracellular permeability. The cytochalasin‐b‐induced disruption of actin microfilaments and subsequent changes in intercellular junctional complexes and paracellular permeability were not affected by inhibitors of protein synthesis (actinomycin‐D or cycloheximide) or microtubule function (colchicine), but were inhibited by metabolic energy inhibitors (2,4‐dinitrophenol or sodium azide). The cytochalasin‐b‐induced disturbance in Caco‐2 actin microfilaments and intercellular junctional complexes and increase in paracellular permeability were rapidly reversed. The paracellular pathway “re‐tightening” following cytochalasin‐b removal was not affected by actinomycin‐D, cycloheximide, or colchicine, but was inhibited by 2,4‐dinitrophenol and sodium azide. The colchicine‐induced disruption of microtubules did not have significant effect on actin microfilaments, intercellular junctions, or paracellular permeability. These findings suggest that cytochalasin‐b‐induced increase in Caco‐2 monolayer paracellular permeability was due to actin microfilament mediated perturbation of intercellular junctional complexes. The re‐tightening of paracellular pathways (following removal of cytochalasin‐b) resulted from energy‐mediated re‐assembly of pre‐existing actin microfilaments and intercellular junctional complexes. This re‐closure process did not require protein synthesis or microtubule‐mediated shuttling process. {\circledC} 1995 Wiley‐Liss, Inc.",
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Cytoskeletal regulation of Caco‐2 intestinal monolayer paracellular permeability. / Ma, Thomas; Hollander, Daniel; Tran, Le Thuy; Nguyen, Don; Hoa, Neil; Bhalla, Deepak.

In: Journal of Cellular Physiology, Vol. 164, No. 3, 01.01.1995, p. 533-545.

Research output: Contribution to journalArticle

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T1 - Cytoskeletal regulation of Caco‐2 intestinal monolayer paracellular permeability

AU - Ma, Thomas

AU - Hollander, Daniel

AU - Tran, Le Thuy

AU - Nguyen, Don

AU - Hoa, Neil

AU - Bhalla, Deepak

PY - 1995/1/1

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N2 - An abnormal increase in intestinal paracellular permeability may be an important pathogenic factor in various intestinal diseases. The intracellular factors and processes that regulate and cause alteration of intestinal paracellular permeability are not well understood. The purpose of this study was to examine some of the intracellular processes involved in cytoskeletal regulation of intestinal epithelial paracellular permeability using the filter‐grown Caco‐2 intestinal epithelial monolayers. Cytochalasin‐b and colchicine were used to disrupt the cytoskeletal elements, actin microfilaments, and microtubules. Cytochalasin‐b (5 m̈g/ml) and colchicine (2 × 10−5M) at the doses used caused marked depolymerization and disruption of actin microfilaments and microtubules, respectively. Cytochalasin‐b‐induced disruption of actin microfilaments resulted in perturbation of tight junctions and desmosomes and an increase in Caco‐2 monolayer paracellular permeability. The cytochalasin‐b‐induced disruption of actin microfilaments and subsequent changes in intercellular junctional complexes and paracellular permeability were not affected by inhibitors of protein synthesis (actinomycin‐D or cycloheximide) or microtubule function (colchicine), but were inhibited by metabolic energy inhibitors (2,4‐dinitrophenol or sodium azide). The cytochalasin‐b‐induced disturbance in Caco‐2 actin microfilaments and intercellular junctional complexes and increase in paracellular permeability were rapidly reversed. The paracellular pathway “re‐tightening” following cytochalasin‐b removal was not affected by actinomycin‐D, cycloheximide, or colchicine, but was inhibited by 2,4‐dinitrophenol and sodium azide. The colchicine‐induced disruption of microtubules did not have significant effect on actin microfilaments, intercellular junctions, or paracellular permeability. These findings suggest that cytochalasin‐b‐induced increase in Caco‐2 monolayer paracellular permeability was due to actin microfilament mediated perturbation of intercellular junctional complexes. The re‐tightening of paracellular pathways (following removal of cytochalasin‐b) resulted from energy‐mediated re‐assembly of pre‐existing actin microfilaments and intercellular junctional complexes. This re‐closure process did not require protein synthesis or microtubule‐mediated shuttling process. © 1995 Wiley‐Liss, Inc.

AB - An abnormal increase in intestinal paracellular permeability may be an important pathogenic factor in various intestinal diseases. The intracellular factors and processes that regulate and cause alteration of intestinal paracellular permeability are not well understood. The purpose of this study was to examine some of the intracellular processes involved in cytoskeletal regulation of intestinal epithelial paracellular permeability using the filter‐grown Caco‐2 intestinal epithelial monolayers. Cytochalasin‐b and colchicine were used to disrupt the cytoskeletal elements, actin microfilaments, and microtubules. Cytochalasin‐b (5 m̈g/ml) and colchicine (2 × 10−5M) at the doses used caused marked depolymerization and disruption of actin microfilaments and microtubules, respectively. Cytochalasin‐b‐induced disruption of actin microfilaments resulted in perturbation of tight junctions and desmosomes and an increase in Caco‐2 monolayer paracellular permeability. The cytochalasin‐b‐induced disruption of actin microfilaments and subsequent changes in intercellular junctional complexes and paracellular permeability were not affected by inhibitors of protein synthesis (actinomycin‐D or cycloheximide) or microtubule function (colchicine), but were inhibited by metabolic energy inhibitors (2,4‐dinitrophenol or sodium azide). The cytochalasin‐b‐induced disturbance in Caco‐2 actin microfilaments and intercellular junctional complexes and increase in paracellular permeability were rapidly reversed. The paracellular pathway “re‐tightening” following cytochalasin‐b removal was not affected by actinomycin‐D, cycloheximide, or colchicine, but was inhibited by 2,4‐dinitrophenol and sodium azide. The colchicine‐induced disruption of microtubules did not have significant effect on actin microfilaments, intercellular junctions, or paracellular permeability. These findings suggest that cytochalasin‐b‐induced increase in Caco‐2 monolayer paracellular permeability was due to actin microfilament mediated perturbation of intercellular junctional complexes. The re‐tightening of paracellular pathways (following removal of cytochalasin‐b) resulted from energy‐mediated re‐assembly of pre‐existing actin microfilaments and intercellular junctional complexes. This re‐closure process did not require protein synthesis or microtubule‐mediated shuttling process. © 1995 Wiley‐Liss, Inc.

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