Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes

Gyu Hwan Park, Jae Ryun Ryu, Chan Young Shin, Min Sik Choi, Byoung H. Han, Won-Ki Kim, Hyoung Chun Kim, Ho K. Kwang

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Serine proteases such as thrombin and trypsin play a key role in the development and repair processes in the central nervous system. Molecular actions of serine proteases include multiple cellular events like activation of protease-activated receptors (PARs). PARs belong to a family of G protein-coupled receptors that can be stimulated through their proteolytic cleavage by ligands. PAR-2 has been implicated in neurodegenerative diseases including astrogliosis. Although recent studies have shown that low concentration of trypsin activates PAR-2, its role in morphological changes in primary astrocytes has not been studied. In the present study, we investigated the effects of PAR-2 in astrocyte stellation in rat primary astrocyte culture. Both trypsin (0.1-1 U/ml) and a PAR-2-activating peptide SLIGRL-NH2 (1-50 μM) significantly reversed the stellation induced by serum deprivation in rat astrocytes. Treatment of astrocytes with trypsin or SLIGRL-NH2 resulted in a transient rise of the intracellular Ca2+ level and trypsin-induced morphological changes were blocked by BAPTA, a Ca2+ chelator. In addition, a protein kinase C (PKC) inhibitor, bisindolylmaleimide significantly inhibited the trypsin-induced morphological changes, whereas activation of PKC by phorbol-12-myristate-13-acetate acted as trypsin. Taken together, these results suggest that activation of PAR-2 by trypsin caused reversal of stellation in cultured astrocytes, in part, via the mobilization of intracellular Ca2+ and activation of PKC.

Original languageEnglish
Pages (from-to)15-23
Number of pages9
JournalNeuroscience Research
Volume54
Issue number1
DOIs
Publication statusPublished - 2006 Jan 1
Externally publishedYes

Fingerprint

PAR-2 Receptor
Astrocytes
Trypsin
Proteinase-Activated Receptors
Protein Kinase C
Serine Proteases
Protein C Inhibitor
Protein Kinase Inhibitors
Chelating Agents
G-Protein-Coupled Receptors
Thrombin
Neurodegenerative Diseases
Acetates
Central Nervous System
Ligands
Peptides

Keywords

  • Ca
  • Protease-activated receptor-2
  • Protein kinase C
  • Rat primary astrocyte cultures
  • Reversal of stellation
  • Trypsin

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes. / Park, Gyu Hwan; Ryu, Jae Ryun; Shin, Chan Young; Choi, Min Sik; Han, Byoung H.; Kim, Won-Ki; Kim, Hyoung Chun; Kwang, Ho K.

In: Neuroscience Research, Vol. 54, No. 1, 01.01.2006, p. 15-23.

Research output: Contribution to journalArticle

Park, Gyu Hwan ; Ryu, Jae Ryun ; Shin, Chan Young ; Choi, Min Sik ; Han, Byoung H. ; Kim, Won-Ki ; Kim, Hyoung Chun ; Kwang, Ho K. / Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes. In: Neuroscience Research. 2006 ; Vol. 54, No. 1. pp. 15-23.
@article{2c7195024bb44843a9bb7382cd515e11,
title = "Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes",
abstract = "Serine proteases such as thrombin and trypsin play a key role in the development and repair processes in the central nervous system. Molecular actions of serine proteases include multiple cellular events like activation of protease-activated receptors (PARs). PARs belong to a family of G protein-coupled receptors that can be stimulated through their proteolytic cleavage by ligands. PAR-2 has been implicated in neurodegenerative diseases including astrogliosis. Although recent studies have shown that low concentration of trypsin activates PAR-2, its role in morphological changes in primary astrocytes has not been studied. In the present study, we investigated the effects of PAR-2 in astrocyte stellation in rat primary astrocyte culture. Both trypsin (0.1-1 U/ml) and a PAR-2-activating peptide SLIGRL-NH2 (1-50 μM) significantly reversed the stellation induced by serum deprivation in rat astrocytes. Treatment of astrocytes with trypsin or SLIGRL-NH2 resulted in a transient rise of the intracellular Ca2+ level and trypsin-induced morphological changes were blocked by BAPTA, a Ca2+ chelator. In addition, a protein kinase C (PKC) inhibitor, bisindolylmaleimide significantly inhibited the trypsin-induced morphological changes, whereas activation of PKC by phorbol-12-myristate-13-acetate acted as trypsin. Taken together, these results suggest that activation of PAR-2 by trypsin caused reversal of stellation in cultured astrocytes, in part, via the mobilization of intracellular Ca2+ and activation of PKC.",
keywords = "Ca, Protease-activated receptor-2, Protein kinase C, Rat primary astrocyte cultures, Reversal of stellation, Trypsin",
author = "Park, {Gyu Hwan} and Ryu, {Jae Ryun} and Shin, {Chan Young} and Choi, {Min Sik} and Han, {Byoung H.} and Won-Ki Kim and Kim, {Hyoung Chun} and Kwang, {Ho K.}",
year = "2006",
month = "1",
day = "1",
doi = "10.1016/j.neures.2005.09.007",
language = "English",
volume = "54",
pages = "15--23",
journal = "Neuroscience Research",
issn = "0168-0102",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes

AU - Park, Gyu Hwan

AU - Ryu, Jae Ryun

AU - Shin, Chan Young

AU - Choi, Min Sik

AU - Han, Byoung H.

AU - Kim, Won-Ki

AU - Kim, Hyoung Chun

AU - Kwang, Ho K.

PY - 2006/1/1

Y1 - 2006/1/1

N2 - Serine proteases such as thrombin and trypsin play a key role in the development and repair processes in the central nervous system. Molecular actions of serine proteases include multiple cellular events like activation of protease-activated receptors (PARs). PARs belong to a family of G protein-coupled receptors that can be stimulated through their proteolytic cleavage by ligands. PAR-2 has been implicated in neurodegenerative diseases including astrogliosis. Although recent studies have shown that low concentration of trypsin activates PAR-2, its role in morphological changes in primary astrocytes has not been studied. In the present study, we investigated the effects of PAR-2 in astrocyte stellation in rat primary astrocyte culture. Both trypsin (0.1-1 U/ml) and a PAR-2-activating peptide SLIGRL-NH2 (1-50 μM) significantly reversed the stellation induced by serum deprivation in rat astrocytes. Treatment of astrocytes with trypsin or SLIGRL-NH2 resulted in a transient rise of the intracellular Ca2+ level and trypsin-induced morphological changes were blocked by BAPTA, a Ca2+ chelator. In addition, a protein kinase C (PKC) inhibitor, bisindolylmaleimide significantly inhibited the trypsin-induced morphological changes, whereas activation of PKC by phorbol-12-myristate-13-acetate acted as trypsin. Taken together, these results suggest that activation of PAR-2 by trypsin caused reversal of stellation in cultured astrocytes, in part, via the mobilization of intracellular Ca2+ and activation of PKC.

AB - Serine proteases such as thrombin and trypsin play a key role in the development and repair processes in the central nervous system. Molecular actions of serine proteases include multiple cellular events like activation of protease-activated receptors (PARs). PARs belong to a family of G protein-coupled receptors that can be stimulated through their proteolytic cleavage by ligands. PAR-2 has been implicated in neurodegenerative diseases including astrogliosis. Although recent studies have shown that low concentration of trypsin activates PAR-2, its role in morphological changes in primary astrocytes has not been studied. In the present study, we investigated the effects of PAR-2 in astrocyte stellation in rat primary astrocyte culture. Both trypsin (0.1-1 U/ml) and a PAR-2-activating peptide SLIGRL-NH2 (1-50 μM) significantly reversed the stellation induced by serum deprivation in rat astrocytes. Treatment of astrocytes with trypsin or SLIGRL-NH2 resulted in a transient rise of the intracellular Ca2+ level and trypsin-induced morphological changes were blocked by BAPTA, a Ca2+ chelator. In addition, a protein kinase C (PKC) inhibitor, bisindolylmaleimide significantly inhibited the trypsin-induced morphological changes, whereas activation of PKC by phorbol-12-myristate-13-acetate acted as trypsin. Taken together, these results suggest that activation of PAR-2 by trypsin caused reversal of stellation in cultured astrocytes, in part, via the mobilization of intracellular Ca2+ and activation of PKC.

KW - Ca

KW - Protease-activated receptor-2

KW - Protein kinase C

KW - Rat primary astrocyte cultures

KW - Reversal of stellation

KW - Trypsin

UR - http://www.scopus.com/inward/record.url?scp=28944453944&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28944453944&partnerID=8YFLogxK

U2 - 10.1016/j.neures.2005.09.007

DO - 10.1016/j.neures.2005.09.007

M3 - Article

VL - 54

SP - 15

EP - 23

JO - Neuroscience Research

JF - Neuroscience Research

SN - 0168-0102

IS - 1

ER -