Injury-induced expression of endothelial nitric oxide synthase by glial and microglial cells in the leech central nervous system within minutes after injury

O. T. Shafer, A. Chen, S. M. Kumar, Kenneth J Muller, C. L. Sahley

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

It is known that nitric oxide (NO) is produced by injured tissues of the mammalian central nervous system (CNS) within days of injury. The aim of the present experiments was to determine the cellular synthesis of NO in the CNS immediately after injury, using the CNS of the leech which is capable of synapse regeneration, as a step towards understanding the role of NO in nerve repair. We report that within minutes after crushing the nerve cord of the leech, the region of damage stained histochemically for NADPH diaphorase, which is indicative of nitric oxide synthase (NOS) activity and was immunoreactive for endothelial NOS (eNOS). On immunoblots of leech CNS extract, the same antibody detected a band with a relative molecular mass of 140,000, which is approximately the size of vertebrate eNOS. Cells expressing eNOS immunoreactivity as a result of injury were identified after freezing nerve cords, a procedure that produced less tissue distortion than mechanical crushing. Immunoreactive cells included connective glia and some microglia. Calmodulin was necessary for the eNOS immunoreactivity: it was blocked by calmodulin antagonist W7 (25 μM), but not by similar concentrations of the less potent calmodulin antagonist W12. Thus in the leech CNS, in which axon and synapse regeneration is successful, an increase in NOS activity at lesions appears to be among the earliest responses to injury and may be important for repair of axons.

Original languageEnglish
Pages (from-to)2171-2175
Number of pages5
JournalProceedings of the Royal Society B: Biological Sciences
Volume265
Issue number1411
StatePublished - Nov 22 1998
Externally publishedYes

Fingerprint

Leeches
leech
Hirudinea
Nitric Oxide Synthase Type III
neuroglia
Neurology
nitric oxide
nervous system
Neuroglia
central nervous system
Central Nervous System
calmodulin
Calmodulin
nitric oxide synthase
Wounds and Injuries
Nitric Oxide
nerve tissue
crushing
Crushing
synapse

Keywords

  • Calmodulin antagonists
  • Endothelial nitric oxide synthase
  • Hirudo medicinalis
  • Nerve injury
  • Nitric oxide
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Agricultural and Biological Sciences (miscellaneous)

Cite this

Injury-induced expression of endothelial nitric oxide synthase by glial and microglial cells in the leech central nervous system within minutes after injury. / Shafer, O. T.; Chen, A.; Kumar, S. M.; Muller, Kenneth J; Sahley, C. L.

In: Proceedings of the Royal Society B: Biological Sciences, Vol. 265, No. 1411, 22.11.1998, p. 2171-2175.

Research output: Contribution to journalArticle

@article{5492a77040c64fe3a3bafde99fd5277c,
title = "Injury-induced expression of endothelial nitric oxide synthase by glial and microglial cells in the leech central nervous system within minutes after injury",
abstract = "It is known that nitric oxide (NO) is produced by injured tissues of the mammalian central nervous system (CNS) within days of injury. The aim of the present experiments was to determine the cellular synthesis of NO in the CNS immediately after injury, using the CNS of the leech which is capable of synapse regeneration, as a step towards understanding the role of NO in nerve repair. We report that within minutes after crushing the nerve cord of the leech, the region of damage stained histochemically for NADPH diaphorase, which is indicative of nitric oxide synthase (NOS) activity and was immunoreactive for endothelial NOS (eNOS). On immunoblots of leech CNS extract, the same antibody detected a band with a relative molecular mass of 140,000, which is approximately the size of vertebrate eNOS. Cells expressing eNOS immunoreactivity as a result of injury were identified after freezing nerve cords, a procedure that produced less tissue distortion than mechanical crushing. Immunoreactive cells included connective glia and some microglia. Calmodulin was necessary for the eNOS immunoreactivity: it was blocked by calmodulin antagonist W7 (25 μM), but not by similar concentrations of the less potent calmodulin antagonist W12. Thus in the leech CNS, in which axon and synapse regeneration is successful, an increase in NOS activity at lesions appears to be among the earliest responses to injury and may be important for repair of axons.",
keywords = "Calmodulin antagonists, Endothelial nitric oxide synthase, Hirudo medicinalis, Nerve injury, Nitric oxide, Nitric oxide synthase",
author = "Shafer, {O. T.} and A. Chen and Kumar, {S. M.} and Muller, {Kenneth J} and Sahley, {C. L.}",
year = "1998",
month = "11",
day = "22",
language = "English",
volume = "265",
pages = "2171--2175",
journal = "Proceedings of the Royal Society B: Biological Sciences",
issn = "0800-4622",
publisher = "Royal Society of London",
number = "1411",

}

TY - JOUR

T1 - Injury-induced expression of endothelial nitric oxide synthase by glial and microglial cells in the leech central nervous system within minutes after injury

AU - Shafer, O. T.

AU - Chen, A.

AU - Kumar, S. M.

AU - Muller, Kenneth J

AU - Sahley, C. L.

PY - 1998/11/22

Y1 - 1998/11/22

N2 - It is known that nitric oxide (NO) is produced by injured tissues of the mammalian central nervous system (CNS) within days of injury. The aim of the present experiments was to determine the cellular synthesis of NO in the CNS immediately after injury, using the CNS of the leech which is capable of synapse regeneration, as a step towards understanding the role of NO in nerve repair. We report that within minutes after crushing the nerve cord of the leech, the region of damage stained histochemically for NADPH diaphorase, which is indicative of nitric oxide synthase (NOS) activity and was immunoreactive for endothelial NOS (eNOS). On immunoblots of leech CNS extract, the same antibody detected a band with a relative molecular mass of 140,000, which is approximately the size of vertebrate eNOS. Cells expressing eNOS immunoreactivity as a result of injury were identified after freezing nerve cords, a procedure that produced less tissue distortion than mechanical crushing. Immunoreactive cells included connective glia and some microglia. Calmodulin was necessary for the eNOS immunoreactivity: it was blocked by calmodulin antagonist W7 (25 μM), but not by similar concentrations of the less potent calmodulin antagonist W12. Thus in the leech CNS, in which axon and synapse regeneration is successful, an increase in NOS activity at lesions appears to be among the earliest responses to injury and may be important for repair of axons.

AB - It is known that nitric oxide (NO) is produced by injured tissues of the mammalian central nervous system (CNS) within days of injury. The aim of the present experiments was to determine the cellular synthesis of NO in the CNS immediately after injury, using the CNS of the leech which is capable of synapse regeneration, as a step towards understanding the role of NO in nerve repair. We report that within minutes after crushing the nerve cord of the leech, the region of damage stained histochemically for NADPH diaphorase, which is indicative of nitric oxide synthase (NOS) activity and was immunoreactive for endothelial NOS (eNOS). On immunoblots of leech CNS extract, the same antibody detected a band with a relative molecular mass of 140,000, which is approximately the size of vertebrate eNOS. Cells expressing eNOS immunoreactivity as a result of injury were identified after freezing nerve cords, a procedure that produced less tissue distortion than mechanical crushing. Immunoreactive cells included connective glia and some microglia. Calmodulin was necessary for the eNOS immunoreactivity: it was blocked by calmodulin antagonist W7 (25 μM), but not by similar concentrations of the less potent calmodulin antagonist W12. Thus in the leech CNS, in which axon and synapse regeneration is successful, an increase in NOS activity at lesions appears to be among the earliest responses to injury and may be important for repair of axons.

KW - Calmodulin antagonists

KW - Endothelial nitric oxide synthase

KW - Hirudo medicinalis

KW - Nerve injury

KW - Nitric oxide

KW - Nitric oxide synthase

UR - http://www.scopus.com/inward/record.url?scp=0032558925&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032558925&partnerID=8YFLogxK

M3 - Article

C2 - 9872006

AN - SCOPUS:0032558925

VL - 265

SP - 2171

EP - 2175

JO - Proceedings of the Royal Society B: Biological Sciences

JF - Proceedings of the Royal Society B: Biological Sciences

SN - 0800-4622

IS - 1411

ER -