Versican localizes to the nucleus in proliferating mesenchymal cells

Jon M. Carthy, Thomas Abraham, Anna J. Meredith, Seti Boroomand, Bruce M. McManus

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Objective Versican is a versatile and highly interactive chondroitin sulfate proteoglycan that is found in the extracellular matrix (ECM) of many tissues and is a major component of developing and developed lesions in atherosclerotic vascular disease. In this paper, we present data to indicate that versican may have important intracellular functions in addition to its better known roles in the ECM. Methods and Results Rat aortic smooth muscle cells were fixed and immunostained for versican and images of fluorescently labeled cells were obtained by confocal microscopy. Intracellular versican was detected in the nucleus and cytosol of vascular smooth muscle cells. The use of a synthetic neutralizing peptide eliminated versican immunostaining, demonstrating the specificity of the antibody used in this study. Western blot of pure nuclear extracts confirmed the presence of versican in the nucleus, and multifluorescent immunostaining showed strong colocalization of versican and nucleolin, suggesting a nucleolar localization of versican in nondividing cells. In dividing valve interstitial cells, a strong signal for versican was observed in and around the condensed chromosomes during the various stages of mitosis. Multifluorescent immunostaining for versican and tubulin revealed versican aggregated at opposing poles of the mitotic spindle during metaphase. Knockdown of versican expression using siRNA disrupted the organization of the mitotic spindle and led to the formation of multipolar spindles during metaphase. Conclusions Collectively, these data suggest an intracellular function for versican in vascular cells where it appears to play a role in mitotic spindle organization during cell division. These observations open a new avenue for studies of versican, suggesting even more diverse roles in vascular health and disease.

Original languageEnglish (US)
Pages (from-to)368-374
Number of pages7
JournalCardiovascular Pathology
Volume24
Issue number6
DOIs
StatePublished - Nov 1 2015

Fingerprint

Versicans
Spindle Apparatus
Metaphase
Vascular Diseases
Smooth Muscle Myocytes
Extracellular Matrix
Chondroitin Sulfate Proteoglycans
Antibody Specificity
Tubulin

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Cardiology and Cardiovascular Medicine

Cite this

Carthy, Jon M. ; Abraham, Thomas ; Meredith, Anna J. ; Boroomand, Seti ; McManus, Bruce M. / Versican localizes to the nucleus in proliferating mesenchymal cells. In: Cardiovascular Pathology. 2015 ; Vol. 24, No. 6. pp. 368-374.
@article{157c1f9347944550a5acfbab48b89b34,
title = "Versican localizes to the nucleus in proliferating mesenchymal cells",
abstract = "Objective Versican is a versatile and highly interactive chondroitin sulfate proteoglycan that is found in the extracellular matrix (ECM) of many tissues and is a major component of developing and developed lesions in atherosclerotic vascular disease. In this paper, we present data to indicate that versican may have important intracellular functions in addition to its better known roles in the ECM. Methods and Results Rat aortic smooth muscle cells were fixed and immunostained for versican and images of fluorescently labeled cells were obtained by confocal microscopy. Intracellular versican was detected in the nucleus and cytosol of vascular smooth muscle cells. The use of a synthetic neutralizing peptide eliminated versican immunostaining, demonstrating the specificity of the antibody used in this study. Western blot of pure nuclear extracts confirmed the presence of versican in the nucleus, and multifluorescent immunostaining showed strong colocalization of versican and nucleolin, suggesting a nucleolar localization of versican in nondividing cells. In dividing valve interstitial cells, a strong signal for versican was observed in and around the condensed chromosomes during the various stages of mitosis. Multifluorescent immunostaining for versican and tubulin revealed versican aggregated at opposing poles of the mitotic spindle during metaphase. Knockdown of versican expression using siRNA disrupted the organization of the mitotic spindle and led to the formation of multipolar spindles during metaphase. Conclusions Collectively, these data suggest an intracellular function for versican in vascular cells where it appears to play a role in mitotic spindle organization during cell division. These observations open a new avenue for studies of versican, suggesting even more diverse roles in vascular health and disease.",
author = "Carthy, {Jon M.} and Thomas Abraham and Meredith, {Anna J.} and Seti Boroomand and McManus, {Bruce M.}",
year = "2015",
month = "11",
day = "1",
doi = "10.1016/j.carpath.2015.07.010",
language = "English (US)",
volume = "24",
pages = "368--374",
journal = "Cardiovascular Pathology",
issn = "1054-8807",
publisher = "Elsevier Inc.",
number = "6",

}

Versican localizes to the nucleus in proliferating mesenchymal cells. / Carthy, Jon M.; Abraham, Thomas; Meredith, Anna J.; Boroomand, Seti; McManus, Bruce M.

In: Cardiovascular Pathology, Vol. 24, No. 6, 01.11.2015, p. 368-374.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Versican localizes to the nucleus in proliferating mesenchymal cells

AU - Carthy, Jon M.

AU - Abraham, Thomas

AU - Meredith, Anna J.

AU - Boroomand, Seti

AU - McManus, Bruce M.

PY - 2015/11/1

Y1 - 2015/11/1

N2 - Objective Versican is a versatile and highly interactive chondroitin sulfate proteoglycan that is found in the extracellular matrix (ECM) of many tissues and is a major component of developing and developed lesions in atherosclerotic vascular disease. In this paper, we present data to indicate that versican may have important intracellular functions in addition to its better known roles in the ECM. Methods and Results Rat aortic smooth muscle cells were fixed and immunostained for versican and images of fluorescently labeled cells were obtained by confocal microscopy. Intracellular versican was detected in the nucleus and cytosol of vascular smooth muscle cells. The use of a synthetic neutralizing peptide eliminated versican immunostaining, demonstrating the specificity of the antibody used in this study. Western blot of pure nuclear extracts confirmed the presence of versican in the nucleus, and multifluorescent immunostaining showed strong colocalization of versican and nucleolin, suggesting a nucleolar localization of versican in nondividing cells. In dividing valve interstitial cells, a strong signal for versican was observed in and around the condensed chromosomes during the various stages of mitosis. Multifluorescent immunostaining for versican and tubulin revealed versican aggregated at opposing poles of the mitotic spindle during metaphase. Knockdown of versican expression using siRNA disrupted the organization of the mitotic spindle and led to the formation of multipolar spindles during metaphase. Conclusions Collectively, these data suggest an intracellular function for versican in vascular cells where it appears to play a role in mitotic spindle organization during cell division. These observations open a new avenue for studies of versican, suggesting even more diverse roles in vascular health and disease.

AB - Objective Versican is a versatile and highly interactive chondroitin sulfate proteoglycan that is found in the extracellular matrix (ECM) of many tissues and is a major component of developing and developed lesions in atherosclerotic vascular disease. In this paper, we present data to indicate that versican may have important intracellular functions in addition to its better known roles in the ECM. Methods and Results Rat aortic smooth muscle cells were fixed and immunostained for versican and images of fluorescently labeled cells were obtained by confocal microscopy. Intracellular versican was detected in the nucleus and cytosol of vascular smooth muscle cells. The use of a synthetic neutralizing peptide eliminated versican immunostaining, demonstrating the specificity of the antibody used in this study. Western blot of pure nuclear extracts confirmed the presence of versican in the nucleus, and multifluorescent immunostaining showed strong colocalization of versican and nucleolin, suggesting a nucleolar localization of versican in nondividing cells. In dividing valve interstitial cells, a strong signal for versican was observed in and around the condensed chromosomes during the various stages of mitosis. Multifluorescent immunostaining for versican and tubulin revealed versican aggregated at opposing poles of the mitotic spindle during metaphase. Knockdown of versican expression using siRNA disrupted the organization of the mitotic spindle and led to the formation of multipolar spindles during metaphase. Conclusions Collectively, these data suggest an intracellular function for versican in vascular cells where it appears to play a role in mitotic spindle organization during cell division. These observations open a new avenue for studies of versican, suggesting even more diverse roles in vascular health and disease.

UR - http://www.scopus.com/inward/record.url?scp=84946472285&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84946472285&partnerID=8YFLogxK

U2 - 10.1016/j.carpath.2015.07.010

DO - 10.1016/j.carpath.2015.07.010

M3 - Article

C2 - 26395512

AN - SCOPUS:84946472285

VL - 24

SP - 368

EP - 374

JO - Cardiovascular Pathology

JF - Cardiovascular Pathology

SN - 1054-8807

IS - 6

ER -