Phenotypic consequences of transforming growth factor β1 gene ablation in murine embryonic fibroblasts: Autocrine control of cell proliferation and extracellular matrix biosynthesis

Chitra Sudarshan, Linda Yaswen, Ashok Kulkarni, Rajendra Raghow

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The profound effects of transforming growth factor β1 (TGF-β1) on the immune system, cardiogenesis, in yolk sac hematopoeisis and in differentiation of endothelium have been demonstrated by detailed analyses of TGF-β1 knockout mice during embryogenesis. We have systematically examined the autocrine and paracrine roles of TGF-β1 in cell proliferation and in its ability to modulate the gene expression of selected components of extracellular matrix (ECM) using embryonic fibroblasts from TGF-β1 null mice (TGF-β1(-/-)). The rates of cell proliferation of embryonic fibroblasts from normal mice (TGF-β1(+/+)) and TGF-β1 null mice were compared by cell counting, by 3H thymidine incorporation, and by measuring the fraction of cells in the G1, S, and G2/M phases of the cell cycle by fluorescent activated cell sorting (FACS). Concurrently, the expression of pro-α(I) collagen, fibronectin, and plasminogen activator inhibitor-1 (PAI-1) was also quantified by hybridization of total mRNA from TGF-β1(+/+) and TGF-β1(-/-) embryonic fibroblasts. We report that TGF-β1(-/-) cells proliferated at about twice the rate of TGF-β1(+/+) cells. Further, TGF-β1 null fibroblasts accumulated and synthesized lower constitutive levels of pro-α1(I) collagen, fibronectin, and PAI-1 mRNA. The quantitative differences in the rates of cell proliferation and ECM gene expression between TGF-β1(+/+) and TGF- β1(-/-) cells could be eliminated by treatment of TGF-β1(+/+) cells with a neutralizing antibody of TGF-β1. Thus, our results are consistent with the hypothesis that TGF-β1 acts as a negative autocrine regulator of growth and a positive autocrine regulator of ECM biosynthesis in embryonic fibroblasts.

Original languageEnglish (US)
Pages (from-to)67-75
Number of pages9
JournalJournal of Cellular Physiology
Volume176
Issue number1
DOIs
StatePublished - Jul 1 1998

Fingerprint

Biosynthesis
Cell proliferation
Transforming Growth Factors
Fibroblasts
Ablation
Extracellular Matrix
Genes
Cell Proliferation
Plasminogen Activator Inhibitor 1
Fibronectins
Gene expression
Collagen
Cells
Gene Expression
Fibroblast Growth Factor 1
Yolk Sac
Messenger RNA
Immune system
G2 Phase
Neutralizing Antibodies

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

@article{39ea8c89449641969cf49d4448b50943,
title = "Phenotypic consequences of transforming growth factor β1 gene ablation in murine embryonic fibroblasts: Autocrine control of cell proliferation and extracellular matrix biosynthesis",
abstract = "The profound effects of transforming growth factor β1 (TGF-β1) on the immune system, cardiogenesis, in yolk sac hematopoeisis and in differentiation of endothelium have been demonstrated by detailed analyses of TGF-β1 knockout mice during embryogenesis. We have systematically examined the autocrine and paracrine roles of TGF-β1 in cell proliferation and in its ability to modulate the gene expression of selected components of extracellular matrix (ECM) using embryonic fibroblasts from TGF-β1 null mice (TGF-β1(-/-)). The rates of cell proliferation of embryonic fibroblasts from normal mice (TGF-β1(+/+)) and TGF-β1 null mice were compared by cell counting, by 3H thymidine incorporation, and by measuring the fraction of cells in the G1, S, and G2/M phases of the cell cycle by fluorescent activated cell sorting (FACS). Concurrently, the expression of pro-α(I) collagen, fibronectin, and plasminogen activator inhibitor-1 (PAI-1) was also quantified by hybridization of total mRNA from TGF-β1(+/+) and TGF-β1(-/-) embryonic fibroblasts. We report that TGF-β1(-/-) cells proliferated at about twice the rate of TGF-β1(+/+) cells. Further, TGF-β1 null fibroblasts accumulated and synthesized lower constitutive levels of pro-α1(I) collagen, fibronectin, and PAI-1 mRNA. The quantitative differences in the rates of cell proliferation and ECM gene expression between TGF-β1(+/+) and TGF- β1(-/-) cells could be eliminated by treatment of TGF-β1(+/+) cells with a neutralizing antibody of TGF-β1. Thus, our results are consistent with the hypothesis that TGF-β1 acts as a negative autocrine regulator of growth and a positive autocrine regulator of ECM biosynthesis in embryonic fibroblasts.",
author = "Chitra Sudarshan and Linda Yaswen and Ashok Kulkarni and Rajendra Raghow",
year = "1998",
month = "7",
day = "1",
doi = "10.1002/(SICI)1097-4652(199807)176:1<67::AID-JCP8>3.0.CO;2-6",
language = "English (US)",
volume = "176",
pages = "67--75",
journal = "Journal of Cellular Physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "1",

}

TY - JOUR

T1 - Phenotypic consequences of transforming growth factor β1 gene ablation in murine embryonic fibroblasts

T2 - Autocrine control of cell proliferation and extracellular matrix biosynthesis

AU - Sudarshan, Chitra

AU - Yaswen, Linda

AU - Kulkarni, Ashok

AU - Raghow, Rajendra

PY - 1998/7/1

Y1 - 1998/7/1

N2 - The profound effects of transforming growth factor β1 (TGF-β1) on the immune system, cardiogenesis, in yolk sac hematopoeisis and in differentiation of endothelium have been demonstrated by detailed analyses of TGF-β1 knockout mice during embryogenesis. We have systematically examined the autocrine and paracrine roles of TGF-β1 in cell proliferation and in its ability to modulate the gene expression of selected components of extracellular matrix (ECM) using embryonic fibroblasts from TGF-β1 null mice (TGF-β1(-/-)). The rates of cell proliferation of embryonic fibroblasts from normal mice (TGF-β1(+/+)) and TGF-β1 null mice were compared by cell counting, by 3H thymidine incorporation, and by measuring the fraction of cells in the G1, S, and G2/M phases of the cell cycle by fluorescent activated cell sorting (FACS). Concurrently, the expression of pro-α(I) collagen, fibronectin, and plasminogen activator inhibitor-1 (PAI-1) was also quantified by hybridization of total mRNA from TGF-β1(+/+) and TGF-β1(-/-) embryonic fibroblasts. We report that TGF-β1(-/-) cells proliferated at about twice the rate of TGF-β1(+/+) cells. Further, TGF-β1 null fibroblasts accumulated and synthesized lower constitutive levels of pro-α1(I) collagen, fibronectin, and PAI-1 mRNA. The quantitative differences in the rates of cell proliferation and ECM gene expression between TGF-β1(+/+) and TGF- β1(-/-) cells could be eliminated by treatment of TGF-β1(+/+) cells with a neutralizing antibody of TGF-β1. Thus, our results are consistent with the hypothesis that TGF-β1 acts as a negative autocrine regulator of growth and a positive autocrine regulator of ECM biosynthesis in embryonic fibroblasts.

AB - The profound effects of transforming growth factor β1 (TGF-β1) on the immune system, cardiogenesis, in yolk sac hematopoeisis and in differentiation of endothelium have been demonstrated by detailed analyses of TGF-β1 knockout mice during embryogenesis. We have systematically examined the autocrine and paracrine roles of TGF-β1 in cell proliferation and in its ability to modulate the gene expression of selected components of extracellular matrix (ECM) using embryonic fibroblasts from TGF-β1 null mice (TGF-β1(-/-)). The rates of cell proliferation of embryonic fibroblasts from normal mice (TGF-β1(+/+)) and TGF-β1 null mice were compared by cell counting, by 3H thymidine incorporation, and by measuring the fraction of cells in the G1, S, and G2/M phases of the cell cycle by fluorescent activated cell sorting (FACS). Concurrently, the expression of pro-α(I) collagen, fibronectin, and plasminogen activator inhibitor-1 (PAI-1) was also quantified by hybridization of total mRNA from TGF-β1(+/+) and TGF-β1(-/-) embryonic fibroblasts. We report that TGF-β1(-/-) cells proliferated at about twice the rate of TGF-β1(+/+) cells. Further, TGF-β1 null fibroblasts accumulated and synthesized lower constitutive levels of pro-α1(I) collagen, fibronectin, and PAI-1 mRNA. The quantitative differences in the rates of cell proliferation and ECM gene expression between TGF-β1(+/+) and TGF- β1(-/-) cells could be eliminated by treatment of TGF-β1(+/+) cells with a neutralizing antibody of TGF-β1. Thus, our results are consistent with the hypothesis that TGF-β1 acts as a negative autocrine regulator of growth and a positive autocrine regulator of ECM biosynthesis in embryonic fibroblasts.

UR - http://www.scopus.com/inward/record.url?scp=0031807616&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031807616&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4652(199807)176:1<67::AID-JCP8>3.0.CO;2-6

DO - 10.1002/(SICI)1097-4652(199807)176:1<67::AID-JCP8>3.0.CO;2-6

M3 - Article

C2 - 9618146

AN - SCOPUS:0031807616

VL - 176

SP - 67

EP - 75

JO - Journal of Cellular Physiology

JF - Journal of Cellular Physiology

SN - 0021-9541

IS - 1

ER -