Induction of angiogenic immediate early genes by activation of FP prostanoid receptors in cultured human ciliary smooth muscle cells

Anthony J. Hutchinson, Serena C. Coons, Chih Ling Chou, Wei Xu, W. Daniel Stamer, David F. Woodward, John W Regan

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Purpose: Agonists of the F prostanoid receptor for prostaglandin F exert. exert an ocular hypotensive effect that has been attributed to increased aqueous humor outflow through the uveoscleral pathway. Although tissue remodeling of the ciliary muscle has been described, the signaling mechanisms that link activation of the FP receptor to remodeling of the ciliary muscle are poorly understood. Herein, we describe the identification of novel signaling mechanisms that may contribute to this process. Materials and Methods: Cultures of human ciliary smooth muscle cells were established from fetal eye tissue explants. The cells were validated by their expression of α-smooth muscle-actin and functional FP receptors. Cultures were treated with prostaglandin F and examined for the induction of three immediate early genes related to tissue remodeling using Western blot analysis, quantitative real-time polymerase chain reaction, and reporter gene assays. Results: Human ciliary smooth muscle cells express functional FP receptors whose activation up-regulates the expression of early growth response factor-1 and connective tissue growth factor at the mRNA and protein levels. Prostaglandin F stimulation also increases the protein expression of hypoxia-inducible factor-1α and activates luciferase reporter plasmids under the control of the hypoxia response element. Conclusions: Early growth response factor-1 and hypoxia-inducible factor-1α are important transcriptional activators of downstream genes involved in tissue remodeling and angiogenesis, whereas connective tissue growth factor is a secreted growth factor that also contributes to these processes. Thus, stimulation of FP receptors in human ciliary smooth muscle cells up-regulates the expression of immediate early genes that may coordinate the remodeling of the ciliary muscle, thereby facilitating aqueous outflow.

Original languageEnglish (US)
Pages (from-to)408-418
Number of pages11
JournalCurrent Eye Research
Volume35
Issue number5
DOIs
StatePublished - May 2010

Fingerprint

Immediate-Early Genes
Transcriptional Activation
Smooth Muscle Myocytes
Connective Tissue Growth Factor
Hypoxia-Inducible Factor 1
Intercellular Signaling Peptides and Proteins
Dinoprost
Muscles
Up-Regulation
Aqueous Humor
Prostaglandins F
Response Elements
Luciferases
Reporter Genes
Prostaglandins
Smooth Muscle
Actins
Real-Time Polymerase Chain Reaction
Proteins
Fetus

Keywords

  • Ciliary muscle
  • CTGF
  • EGR-1
  • FP prostanoid receptor
  • HIF-1α
  • Human eye

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Induction of angiogenic immediate early genes by activation of FP prostanoid receptors in cultured human ciliary smooth muscle cells. / Hutchinson, Anthony J.; Coons, Serena C.; Chou, Chih Ling; Xu, Wei; Stamer, W. Daniel; Woodward, David F.; Regan, John W.

In: Current Eye Research, Vol. 35, No. 5, 05.2010, p. 408-418.

Research output: Contribution to journalArticle

Hutchinson, Anthony J. ; Coons, Serena C. ; Chou, Chih Ling ; Xu, Wei ; Stamer, W. Daniel ; Woodward, David F. ; Regan, John W. / Induction of angiogenic immediate early genes by activation of FP prostanoid receptors in cultured human ciliary smooth muscle cells. In: Current Eye Research. 2010 ; Vol. 35, No. 5. pp. 408-418.
@article{a64b69f9e31d4bb182ee2bcb8ff03655,
title = "Induction of angiogenic immediate early genes by activation of FP prostanoid receptors in cultured human ciliary smooth muscle cells",
abstract = "Purpose: Agonists of the F prostanoid receptor for prostaglandin F 2α exert. exert an ocular hypotensive effect that has been attributed to increased aqueous humor outflow through the uveoscleral pathway. Although tissue remodeling of the ciliary muscle has been described, the signaling mechanisms that link activation of the FP receptor to remodeling of the ciliary muscle are poorly understood. Herein, we describe the identification of novel signaling mechanisms that may contribute to this process. Materials and Methods: Cultures of human ciliary smooth muscle cells were established from fetal eye tissue explants. The cells were validated by their expression of α-smooth muscle-actin and functional FP receptors. Cultures were treated with prostaglandin F2α and examined for the induction of three immediate early genes related to tissue remodeling using Western blot analysis, quantitative real-time polymerase chain reaction, and reporter gene assays. Results: Human ciliary smooth muscle cells express functional FP receptors whose activation up-regulates the expression of early growth response factor-1 and connective tissue growth factor at the mRNA and protein levels. Prostaglandin F2α stimulation also increases the protein expression of hypoxia-inducible factor-1α and activates luciferase reporter plasmids under the control of the hypoxia response element. Conclusions: Early growth response factor-1 and hypoxia-inducible factor-1α are important transcriptional activators of downstream genes involved in tissue remodeling and angiogenesis, whereas connective tissue growth factor is a secreted growth factor that also contributes to these processes. Thus, stimulation of FP receptors in human ciliary smooth muscle cells up-regulates the expression of immediate early genes that may coordinate the remodeling of the ciliary muscle, thereby facilitating aqueous outflow.",
keywords = "Ciliary muscle, CTGF, EGR-1, FP prostanoid receptor, HIF-1α, Human eye",
author = "Hutchinson, {Anthony J.} and Coons, {Serena C.} and Chou, {Chih Ling} and Wei Xu and Stamer, {W. Daniel} and Woodward, {David F.} and Regan, {John W}",
year = "2010",
month = "5",
doi = "10.3109/02713680903581021",
language = "English (US)",
volume = "35",
pages = "408--418",
journal = "Current Eye Research",
issn = "0271-3683",
publisher = "Informa Healthcare",
number = "5",

}

TY - JOUR

T1 - Induction of angiogenic immediate early genes by activation of FP prostanoid receptors in cultured human ciliary smooth muscle cells

AU - Hutchinson, Anthony J.

AU - Coons, Serena C.

AU - Chou, Chih Ling

AU - Xu, Wei

AU - Stamer, W. Daniel

AU - Woodward, David F.

AU - Regan, John W

PY - 2010/5

Y1 - 2010/5

N2 - Purpose: Agonists of the F prostanoid receptor for prostaglandin F 2α exert. exert an ocular hypotensive effect that has been attributed to increased aqueous humor outflow through the uveoscleral pathway. Although tissue remodeling of the ciliary muscle has been described, the signaling mechanisms that link activation of the FP receptor to remodeling of the ciliary muscle are poorly understood. Herein, we describe the identification of novel signaling mechanisms that may contribute to this process. Materials and Methods: Cultures of human ciliary smooth muscle cells were established from fetal eye tissue explants. The cells were validated by their expression of α-smooth muscle-actin and functional FP receptors. Cultures were treated with prostaglandin F2α and examined for the induction of three immediate early genes related to tissue remodeling using Western blot analysis, quantitative real-time polymerase chain reaction, and reporter gene assays. Results: Human ciliary smooth muscle cells express functional FP receptors whose activation up-regulates the expression of early growth response factor-1 and connective tissue growth factor at the mRNA and protein levels. Prostaglandin F2α stimulation also increases the protein expression of hypoxia-inducible factor-1α and activates luciferase reporter plasmids under the control of the hypoxia response element. Conclusions: Early growth response factor-1 and hypoxia-inducible factor-1α are important transcriptional activators of downstream genes involved in tissue remodeling and angiogenesis, whereas connective tissue growth factor is a secreted growth factor that also contributes to these processes. Thus, stimulation of FP receptors in human ciliary smooth muscle cells up-regulates the expression of immediate early genes that may coordinate the remodeling of the ciliary muscle, thereby facilitating aqueous outflow.

AB - Purpose: Agonists of the F prostanoid receptor for prostaglandin F 2α exert. exert an ocular hypotensive effect that has been attributed to increased aqueous humor outflow through the uveoscleral pathway. Although tissue remodeling of the ciliary muscle has been described, the signaling mechanisms that link activation of the FP receptor to remodeling of the ciliary muscle are poorly understood. Herein, we describe the identification of novel signaling mechanisms that may contribute to this process. Materials and Methods: Cultures of human ciliary smooth muscle cells were established from fetal eye tissue explants. The cells were validated by their expression of α-smooth muscle-actin and functional FP receptors. Cultures were treated with prostaglandin F2α and examined for the induction of three immediate early genes related to tissue remodeling using Western blot analysis, quantitative real-time polymerase chain reaction, and reporter gene assays. Results: Human ciliary smooth muscle cells express functional FP receptors whose activation up-regulates the expression of early growth response factor-1 and connective tissue growth factor at the mRNA and protein levels. Prostaglandin F2α stimulation also increases the protein expression of hypoxia-inducible factor-1α and activates luciferase reporter plasmids under the control of the hypoxia response element. Conclusions: Early growth response factor-1 and hypoxia-inducible factor-1α are important transcriptional activators of downstream genes involved in tissue remodeling and angiogenesis, whereas connective tissue growth factor is a secreted growth factor that also contributes to these processes. Thus, stimulation of FP receptors in human ciliary smooth muscle cells up-regulates the expression of immediate early genes that may coordinate the remodeling of the ciliary muscle, thereby facilitating aqueous outflow.

KW - Ciliary muscle

KW - CTGF

KW - EGR-1

KW - FP prostanoid receptor

KW - HIF-1α

KW - Human eye

UR - http://www.scopus.com/inward/record.url?scp=77952235066&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77952235066&partnerID=8YFLogxK

U2 - 10.3109/02713680903581021

DO - 10.3109/02713680903581021

M3 - Article

VL - 35

SP - 408

EP - 418

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 5

ER -