Thrombin induced inhibition of neurite outgrowth from dorsal root ganglion neurons

Jagjit S. Gill, Kelly Pitts, Frank M. Rusnak, Whyte G. Owen, Anthony John Windebank

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Thrombin is a multifunctional protease. Recent studies on cultured neuronal cells have suggested a function for thrombin in the development and maintenance of the nervous system. Thrombin has been found to induce neurite retraction and reverse stellation in neuroblastoma cell lines and rat astrocytes, respectively. The major focus of our study was to investigate the potential role of thrombin in peripheral nervous system development using the rat embryonic dorsal root ganglion model. We found a dose dependent inhibition of neurite outgrowth from explant dorsal root ganglion cultures upon exposure to 2 to 200 nM thrombin. This effect was reversed by the specific thrombin inhibitor, hirudin. A synthetic peptide that imitates the fully active receptor, thrombin receptor activating peptide, was also found to inhibit neurite outgrowth from dorsal root ganglia. bis-Benzimide stained neuronal cultures did not show any evidence of cell death after exposure to thrombin or thrombin receptor activating peptides. Immunohistochemical studies revealed specific staining of the thrombin receptor on neurons, with intense labeling along neurites. Enriched neuronal cultures exposed to thrombin and thrombin receptor activating peptides revealed rapid activation of phospholipase Cγ-1, a second messenger associated with the thrombin receptor. These findings are the first to describe the localization of the thrombin receptor to dorsal root ganglion neurons. We propose that receptor activation is associated with thrombin induced inhibition of neurite outgrowth.

Original languageEnglish (US)
Pages (from-to)321-327
Number of pages7
JournalBrain Research
Volume797
Issue number2
DOIs
StatePublished - Jun 29 1998

Fingerprint

Spinal Ganglia
Thrombin
Thrombin Receptors
Neurons
thrombin receptor peptide SFLLRNP
Neurites
Hirudins
Neuronal Outgrowth
Peripheral Nervous System
Type C Phospholipases
Second Messenger Systems
Neuroblastoma
Astrocytes
Nervous System
Cultured Cells
Peptide Hydrolases
Cell Death
Maintenance
Staining and Labeling
Cell Line

Keywords

  • Hirudin
  • Peripheral nervous system
  • PL Cγ-1
  • Thrombin receptor
  • Thrombin receptor activating peptide

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Thrombin induced inhibition of neurite outgrowth from dorsal root ganglion neurons. / Gill, Jagjit S.; Pitts, Kelly; Rusnak, Frank M.; Owen, Whyte G.; Windebank, Anthony John.

In: Brain Research, Vol. 797, No. 2, 29.06.1998, p. 321-327.

Research output: Contribution to journalArticle

Gill, Jagjit S. ; Pitts, Kelly ; Rusnak, Frank M. ; Owen, Whyte G. ; Windebank, Anthony John. / Thrombin induced inhibition of neurite outgrowth from dorsal root ganglion neurons. In: Brain Research. 1998 ; Vol. 797, No. 2. pp. 321-327.
@article{81480f5faa7d4d6fa81668bbdbe2ce56,
title = "Thrombin induced inhibition of neurite outgrowth from dorsal root ganglion neurons",
abstract = "Thrombin is a multifunctional protease. Recent studies on cultured neuronal cells have suggested a function for thrombin in the development and maintenance of the nervous system. Thrombin has been found to induce neurite retraction and reverse stellation in neuroblastoma cell lines and rat astrocytes, respectively. The major focus of our study was to investigate the potential role of thrombin in peripheral nervous system development using the rat embryonic dorsal root ganglion model. We found a dose dependent inhibition of neurite outgrowth from explant dorsal root ganglion cultures upon exposure to 2 to 200 nM thrombin. This effect was reversed by the specific thrombin inhibitor, hirudin. A synthetic peptide that imitates the fully active receptor, thrombin receptor activating peptide, was also found to inhibit neurite outgrowth from dorsal root ganglia. bis-Benzimide stained neuronal cultures did not show any evidence of cell death after exposure to thrombin or thrombin receptor activating peptides. Immunohistochemical studies revealed specific staining of the thrombin receptor on neurons, with intense labeling along neurites. Enriched neuronal cultures exposed to thrombin and thrombin receptor activating peptides revealed rapid activation of phospholipase Cγ-1, a second messenger associated with the thrombin receptor. These findings are the first to describe the localization of the thrombin receptor to dorsal root ganglion neurons. We propose that receptor activation is associated with thrombin induced inhibition of neurite outgrowth.",
keywords = "Hirudin, Peripheral nervous system, PL Cγ-1, Thrombin receptor, Thrombin receptor activating peptide",
author = "Gill, {Jagjit S.} and Kelly Pitts and Rusnak, {Frank M.} and Owen, {Whyte G.} and Windebank, {Anthony John}",
year = "1998",
month = "6",
day = "29",
doi = "10.1016/S0006-8993(98)00344-8",
language = "English (US)",
volume = "797",
pages = "321--327",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Thrombin induced inhibition of neurite outgrowth from dorsal root ganglion neurons

AU - Gill, Jagjit S.

AU - Pitts, Kelly

AU - Rusnak, Frank M.

AU - Owen, Whyte G.

AU - Windebank, Anthony John

PY - 1998/6/29

Y1 - 1998/6/29

N2 - Thrombin is a multifunctional protease. Recent studies on cultured neuronal cells have suggested a function for thrombin in the development and maintenance of the nervous system. Thrombin has been found to induce neurite retraction and reverse stellation in neuroblastoma cell lines and rat astrocytes, respectively. The major focus of our study was to investigate the potential role of thrombin in peripheral nervous system development using the rat embryonic dorsal root ganglion model. We found a dose dependent inhibition of neurite outgrowth from explant dorsal root ganglion cultures upon exposure to 2 to 200 nM thrombin. This effect was reversed by the specific thrombin inhibitor, hirudin. A synthetic peptide that imitates the fully active receptor, thrombin receptor activating peptide, was also found to inhibit neurite outgrowth from dorsal root ganglia. bis-Benzimide stained neuronal cultures did not show any evidence of cell death after exposure to thrombin or thrombin receptor activating peptides. Immunohistochemical studies revealed specific staining of the thrombin receptor on neurons, with intense labeling along neurites. Enriched neuronal cultures exposed to thrombin and thrombin receptor activating peptides revealed rapid activation of phospholipase Cγ-1, a second messenger associated with the thrombin receptor. These findings are the first to describe the localization of the thrombin receptor to dorsal root ganglion neurons. We propose that receptor activation is associated with thrombin induced inhibition of neurite outgrowth.

AB - Thrombin is a multifunctional protease. Recent studies on cultured neuronal cells have suggested a function for thrombin in the development and maintenance of the nervous system. Thrombin has been found to induce neurite retraction and reverse stellation in neuroblastoma cell lines and rat astrocytes, respectively. The major focus of our study was to investigate the potential role of thrombin in peripheral nervous system development using the rat embryonic dorsal root ganglion model. We found a dose dependent inhibition of neurite outgrowth from explant dorsal root ganglion cultures upon exposure to 2 to 200 nM thrombin. This effect was reversed by the specific thrombin inhibitor, hirudin. A synthetic peptide that imitates the fully active receptor, thrombin receptor activating peptide, was also found to inhibit neurite outgrowth from dorsal root ganglia. bis-Benzimide stained neuronal cultures did not show any evidence of cell death after exposure to thrombin or thrombin receptor activating peptides. Immunohistochemical studies revealed specific staining of the thrombin receptor on neurons, with intense labeling along neurites. Enriched neuronal cultures exposed to thrombin and thrombin receptor activating peptides revealed rapid activation of phospholipase Cγ-1, a second messenger associated with the thrombin receptor. These findings are the first to describe the localization of the thrombin receptor to dorsal root ganglion neurons. We propose that receptor activation is associated with thrombin induced inhibition of neurite outgrowth.

KW - Hirudin

KW - Peripheral nervous system

KW - PL Cγ-1

KW - Thrombin receptor

KW - Thrombin receptor activating peptide

UR - http://www.scopus.com/inward/record.url?scp=0032578046&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032578046&partnerID=8YFLogxK

U2 - 10.1016/S0006-8993(98)00344-8

DO - 10.1016/S0006-8993(98)00344-8

M3 - Article

C2 - 9666159

AN - SCOPUS:0032578046

VL - 797

SP - 321

EP - 327

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 2

ER -