Retarded intracellular lipid transport associated with reduced expression of Cdc42, a member of Rho-GTPases, in human aged skin fibroblasts

A possible function of Cdc42 in mediating intracellular lipid transport

Kosuke Tsukamoto, Ken ichi Hirano, Shizuya Yamashita, Naohiko Sakai, Chiaki Ikegami, Zhongyan Zhang, Fumihiko Matsuura, Hisatoyo Hiraoka, Akifumi Matsuyama, Masato Ishigami, Yuji Matsuzawa

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Objective - Many cell types in atherosclerotic lesions are thought to have various biological abnormalities, such as impaired lipid homeostasis and slow cell proliferation, which may be related to senescence at cellular and individual levels. One of the common characteristics of senescent cells in vitro is the alteration of actin cytoskeletons, which have been reported to be involved in the intracellular transport of lipids. Recently, we raised the hypothesis that Cdc42, which is a member of the Rho-GTPase family and is known to play an important role in actin dynamics, might be important in cellular lipid transport. Methods and Results - In the present study, we found that the protein expression levels and GTP-binding activities of Cdc42 were decreased in aged human skin fibroblasts. Moreover, we found the intracellular kinetics of Golgi-associated lipids to be retarded in these cells, which was demonstrated by the fluorescence recovery after photobleaching (FRAP) technique and the use of N-[7-(4-nitrobenzo-2-oxa-1,3- diazole)]-6-aminohexanoyl-D-erythro-sphingosine as a tracer. To correlate the decreased expression of Cdc42 with the retarded FRAP, we complemented the amount of wild-type c-myc-tagged Cdc42Hs (myc-Cdc42Hs-WT) by adenovirus-mediated gene transfer. We further tested the effect of the dominant-active form (myc-Cdc42Hs-DA, V12Cdc42Hs) or dominant-negative form (myc-Cdc42Hs-DN, N17Cdc42Hs) of Cdc42Hs on FRAP. Introduction of myc-Cdc42Hs-WT or myc-Cdc42Hs-DA recovered the retarded FRAP in the aged fibroblasts. Conversely, control fibroblasts infected with myc-Cdc42Hs-DN exhibited significantly retarded FRAP. Conclusions - These data clearly indicate that the expression of Cdc42, a small G protein, is decreased in the aged cells in close association with the retarded intracellular lipid transport. The present study demonstrates a possible function of Cdc42 in the mediation of intracellular lipid transport.

Original languageEnglish
Pages (from-to)1899-1904
Number of pages6
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume22
Issue number11
DOIs
Publication statusPublished - 01-11-2002

Fingerprint

rho GTP-Binding Proteins
Fluorescence Recovery After Photobleaching
Fibroblasts
Lipids
Skin
Monomeric GTP-Binding Proteins
Cell Aging
Guanosine Triphosphate
Actin Cytoskeleton
Adenoviridae
Actins
Homeostasis
Cell Proliferation
Genes
Proteins

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine

Cite this

Tsukamoto, Kosuke ; Hirano, Ken ichi ; Yamashita, Shizuya ; Sakai, Naohiko ; Ikegami, Chiaki ; Zhang, Zhongyan ; Matsuura, Fumihiko ; Hiraoka, Hisatoyo ; Matsuyama, Akifumi ; Ishigami, Masato ; Matsuzawa, Yuji. / Retarded intracellular lipid transport associated with reduced expression of Cdc42, a member of Rho-GTPases, in human aged skin fibroblasts : A possible function of Cdc42 in mediating intracellular lipid transport. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2002 ; Vol. 22, No. 11. pp. 1899-1904.
@article{010fee92c7624e8da6d5fd2ec0dde041,
title = "Retarded intracellular lipid transport associated with reduced expression of Cdc42, a member of Rho-GTPases, in human aged skin fibroblasts: A possible function of Cdc42 in mediating intracellular lipid transport",
abstract = "Objective - Many cell types in atherosclerotic lesions are thought to have various biological abnormalities, such as impaired lipid homeostasis and slow cell proliferation, which may be related to senescence at cellular and individual levels. One of the common characteristics of senescent cells in vitro is the alteration of actin cytoskeletons, which have been reported to be involved in the intracellular transport of lipids. Recently, we raised the hypothesis that Cdc42, which is a member of the Rho-GTPase family and is known to play an important role in actin dynamics, might be important in cellular lipid transport. Methods and Results - In the present study, we found that the protein expression levels and GTP-binding activities of Cdc42 were decreased in aged human skin fibroblasts. Moreover, we found the intracellular kinetics of Golgi-associated lipids to be retarded in these cells, which was demonstrated by the fluorescence recovery after photobleaching (FRAP) technique and the use of N-[7-(4-nitrobenzo-2-oxa-1,3- diazole)]-6-aminohexanoyl-D-erythro-sphingosine as a tracer. To correlate the decreased expression of Cdc42 with the retarded FRAP, we complemented the amount of wild-type c-myc-tagged Cdc42Hs (myc-Cdc42Hs-WT) by adenovirus-mediated gene transfer. We further tested the effect of the dominant-active form (myc-Cdc42Hs-DA, V12Cdc42Hs) or dominant-negative form (myc-Cdc42Hs-DN, N17Cdc42Hs) of Cdc42Hs on FRAP. Introduction of myc-Cdc42Hs-WT or myc-Cdc42Hs-DA recovered the retarded FRAP in the aged fibroblasts. Conversely, control fibroblasts infected with myc-Cdc42Hs-DN exhibited significantly retarded FRAP. Conclusions - These data clearly indicate that the expression of Cdc42, a small G protein, is decreased in the aged cells in close association with the retarded intracellular lipid transport. The present study demonstrates a possible function of Cdc42 in the mediation of intracellular lipid transport.",
author = "Kosuke Tsukamoto and Hirano, {Ken ichi} and Shizuya Yamashita and Naohiko Sakai and Chiaki Ikegami and Zhongyan Zhang and Fumihiko Matsuura and Hisatoyo Hiraoka and Akifumi Matsuyama and Masato Ishigami and Yuji Matsuzawa",
year = "2002",
month = "11",
day = "1",
doi = "10.1161/01.ATV.0000036080.42391.33",
language = "English",
volume = "22",
pages = "1899--1904",
journal = "Arteriosclerosis, Thrombosis, and Vascular Biology",
issn = "1079-5642",
publisher = "Lippincott Williams and Wilkins",
number = "11",

}

Retarded intracellular lipid transport associated with reduced expression of Cdc42, a member of Rho-GTPases, in human aged skin fibroblasts : A possible function of Cdc42 in mediating intracellular lipid transport. / Tsukamoto, Kosuke; Hirano, Ken ichi; Yamashita, Shizuya; Sakai, Naohiko; Ikegami, Chiaki; Zhang, Zhongyan; Matsuura, Fumihiko; Hiraoka, Hisatoyo; Matsuyama, Akifumi; Ishigami, Masato; Matsuzawa, Yuji.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 22, No. 11, 01.11.2002, p. 1899-1904.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Retarded intracellular lipid transport associated with reduced expression of Cdc42, a member of Rho-GTPases, in human aged skin fibroblasts

T2 - A possible function of Cdc42 in mediating intracellular lipid transport

AU - Tsukamoto, Kosuke

AU - Hirano, Ken ichi

AU - Yamashita, Shizuya

AU - Sakai, Naohiko

AU - Ikegami, Chiaki

AU - Zhang, Zhongyan

AU - Matsuura, Fumihiko

AU - Hiraoka, Hisatoyo

AU - Matsuyama, Akifumi

AU - Ishigami, Masato

AU - Matsuzawa, Yuji

PY - 2002/11/1

Y1 - 2002/11/1

N2 - Objective - Many cell types in atherosclerotic lesions are thought to have various biological abnormalities, such as impaired lipid homeostasis and slow cell proliferation, which may be related to senescence at cellular and individual levels. One of the common characteristics of senescent cells in vitro is the alteration of actin cytoskeletons, which have been reported to be involved in the intracellular transport of lipids. Recently, we raised the hypothesis that Cdc42, which is a member of the Rho-GTPase family and is known to play an important role in actin dynamics, might be important in cellular lipid transport. Methods and Results - In the present study, we found that the protein expression levels and GTP-binding activities of Cdc42 were decreased in aged human skin fibroblasts. Moreover, we found the intracellular kinetics of Golgi-associated lipids to be retarded in these cells, which was demonstrated by the fluorescence recovery after photobleaching (FRAP) technique and the use of N-[7-(4-nitrobenzo-2-oxa-1,3- diazole)]-6-aminohexanoyl-D-erythro-sphingosine as a tracer. To correlate the decreased expression of Cdc42 with the retarded FRAP, we complemented the amount of wild-type c-myc-tagged Cdc42Hs (myc-Cdc42Hs-WT) by adenovirus-mediated gene transfer. We further tested the effect of the dominant-active form (myc-Cdc42Hs-DA, V12Cdc42Hs) or dominant-negative form (myc-Cdc42Hs-DN, N17Cdc42Hs) of Cdc42Hs on FRAP. Introduction of myc-Cdc42Hs-WT or myc-Cdc42Hs-DA recovered the retarded FRAP in the aged fibroblasts. Conversely, control fibroblasts infected with myc-Cdc42Hs-DN exhibited significantly retarded FRAP. Conclusions - These data clearly indicate that the expression of Cdc42, a small G protein, is decreased in the aged cells in close association with the retarded intracellular lipid transport. The present study demonstrates a possible function of Cdc42 in the mediation of intracellular lipid transport.

AB - Objective - Many cell types in atherosclerotic lesions are thought to have various biological abnormalities, such as impaired lipid homeostasis and slow cell proliferation, which may be related to senescence at cellular and individual levels. One of the common characteristics of senescent cells in vitro is the alteration of actin cytoskeletons, which have been reported to be involved in the intracellular transport of lipids. Recently, we raised the hypothesis that Cdc42, which is a member of the Rho-GTPase family and is known to play an important role in actin dynamics, might be important in cellular lipid transport. Methods and Results - In the present study, we found that the protein expression levels and GTP-binding activities of Cdc42 were decreased in aged human skin fibroblasts. Moreover, we found the intracellular kinetics of Golgi-associated lipids to be retarded in these cells, which was demonstrated by the fluorescence recovery after photobleaching (FRAP) technique and the use of N-[7-(4-nitrobenzo-2-oxa-1,3- diazole)]-6-aminohexanoyl-D-erythro-sphingosine as a tracer. To correlate the decreased expression of Cdc42 with the retarded FRAP, we complemented the amount of wild-type c-myc-tagged Cdc42Hs (myc-Cdc42Hs-WT) by adenovirus-mediated gene transfer. We further tested the effect of the dominant-active form (myc-Cdc42Hs-DA, V12Cdc42Hs) or dominant-negative form (myc-Cdc42Hs-DN, N17Cdc42Hs) of Cdc42Hs on FRAP. Introduction of myc-Cdc42Hs-WT or myc-Cdc42Hs-DA recovered the retarded FRAP in the aged fibroblasts. Conversely, control fibroblasts infected with myc-Cdc42Hs-DN exhibited significantly retarded FRAP. Conclusions - These data clearly indicate that the expression of Cdc42, a small G protein, is decreased in the aged cells in close association with the retarded intracellular lipid transport. The present study demonstrates a possible function of Cdc42 in the mediation of intracellular lipid transport.

UR - http://www.scopus.com/inward/record.url?scp=0036846870&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036846870&partnerID=8YFLogxK

U2 - 10.1161/01.ATV.0000036080.42391.33

DO - 10.1161/01.ATV.0000036080.42391.33

M3 - Article

VL - 22

SP - 1899

EP - 1904

JO - Arteriosclerosis, Thrombosis, and Vascular Biology

JF - Arteriosclerosis, Thrombosis, and Vascular Biology

SN - 1079-5642

IS - 11

ER -