Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons

Helen Hellmich, Jeanna M. Garcia, Megumi Shimamura, Syed A. Shah, Marcela A. Avila, Tatsuo Uchida, Margaret A. Parsley, Bridget A. Capra, Kristine A. Eidson, Deborah R. Kennedy, John Winston, Douglas Dewitt, Donald Prough

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: After traumatic brain injury, memory dysfunction is due in part to damage to the hippocampus. To study the molecular mechanisms of this selective vulnerability, the authors used laser capture microdissection of neurons stained with Fluoro-Jade to directly compare gene expression in injured (Fluoro-Jade-positive) and adjacent uninjured (Fluoro-Jade-negative) rat hippocampal neurons after traumatic brain injury and traumatic brain injury plus hemorrhagic hypotension. Methods: Twelve isoflurane-anesthetized Sprague-Dawley rats underwent moderate (2.0 atm) fluid percussion traumatic brain injury followed by either normotenslon or hemorrhagic hypotension. Animals were killed 24 h after injury. Frozen brain sections were double stained with 1% cresyl violet and 0.001% Fluoro-Jade. RNA from 10 Fluoro-Jade-positive neurons and 10 Fluoro-Jade-negative neurons, obtained from the hippocampal CA1, CA3, and dentate gyrus subfields using laser capture microdissection, was linearly amplified and analyzed by quantitative ribonuclease protection assay for nine neuroprotective and apoptosis-related genes. Results: In injured CA3 neurons, expression of the neuroprotective genes glutathaione peroxidase 1, heme oxygenase 1, and brain-derived neurotrophic factor was significantly decreased compared with that of adjacent uninjured neurons. Superimposition of hemorrhagic hypotension was associated with down-regulation of neuroprotective genes in both injured and uninjured neurons of all subregions. Expression of apoptosis-related genes did not vary between injured and uninjured neurons, with or without superimposed hemorrhage. Conclusions: The authors show, in the first direct comparison of messenger RNA levels in injured and uninjured hippocampal neurons, that injured neurons express lower levels of neuroprotective genes than adjacent uninjured neurons.

Original languageEnglish (US)
Pages (from-to)806-814
Number of pages9
JournalAnesthesiology
Volume102
Issue number4
DOIs
StatePublished - Apr 2005

Fingerprint

Hypotension
Gene Expression
Neurons
Laser Capture Microdissection
Genes
Traumatic Brain Injury
Apoptosis
Percussion
Heme Oxygenase-1
Isoflurane
Brain-Derived Neurotrophic Factor
Dentate Gyrus
Frozen Sections
Ribonucleases
Peroxidase
Sprague Dawley Rats
fluoro jade
Hippocampus
Down-Regulation
RNA

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons. / Hellmich, Helen; Garcia, Jeanna M.; Shimamura, Megumi; Shah, Syed A.; Avila, Marcela A.; Uchida, Tatsuo; Parsley, Margaret A.; Capra, Bridget A.; Eidson, Kristine A.; Kennedy, Deborah R.; Winston, John; Dewitt, Douglas; Prough, Donald.

In: Anesthesiology, Vol. 102, No. 4, 04.2005, p. 806-814.

Research output: Contribution to journalArticle

Hellmich, Helen ; Garcia, Jeanna M. ; Shimamura, Megumi ; Shah, Syed A. ; Avila, Marcela A. ; Uchida, Tatsuo ; Parsley, Margaret A. ; Capra, Bridget A. ; Eidson, Kristine A. ; Kennedy, Deborah R. ; Winston, John ; Dewitt, Douglas ; Prough, Donald. / Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons. In: Anesthesiology. 2005 ; Vol. 102, No. 4. pp. 806-814.
@article{bdfe028ca183428284355f2f8ea5ffb2,
title = "Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons",
abstract = "Background: After traumatic brain injury, memory dysfunction is due in part to damage to the hippocampus. To study the molecular mechanisms of this selective vulnerability, the authors used laser capture microdissection of neurons stained with Fluoro-Jade to directly compare gene expression in injured (Fluoro-Jade-positive) and adjacent uninjured (Fluoro-Jade-negative) rat hippocampal neurons after traumatic brain injury and traumatic brain injury plus hemorrhagic hypotension. Methods: Twelve isoflurane-anesthetized Sprague-Dawley rats underwent moderate (2.0 atm) fluid percussion traumatic brain injury followed by either normotenslon or hemorrhagic hypotension. Animals were killed 24 h after injury. Frozen brain sections were double stained with 1{\%} cresyl violet and 0.001{\%} Fluoro-Jade. RNA from 10 Fluoro-Jade-positive neurons and 10 Fluoro-Jade-negative neurons, obtained from the hippocampal CA1, CA3, and dentate gyrus subfields using laser capture microdissection, was linearly amplified and analyzed by quantitative ribonuclease protection assay for nine neuroprotective and apoptosis-related genes. Results: In injured CA3 neurons, expression of the neuroprotective genes glutathaione peroxidase 1, heme oxygenase 1, and brain-derived neurotrophic factor was significantly decreased compared with that of adjacent uninjured neurons. Superimposition of hemorrhagic hypotension was associated with down-regulation of neuroprotective genes in both injured and uninjured neurons of all subregions. Expression of apoptosis-related genes did not vary between injured and uninjured neurons, with or without superimposed hemorrhage. Conclusions: The authors show, in the first direct comparison of messenger RNA levels in injured and uninjured hippocampal neurons, that injured neurons express lower levels of neuroprotective genes than adjacent uninjured neurons.",
author = "Helen Hellmich and Garcia, {Jeanna M.} and Megumi Shimamura and Shah, {Syed A.} and Avila, {Marcela A.} and Tatsuo Uchida and Parsley, {Margaret A.} and Capra, {Bridget A.} and Eidson, {Kristine A.} and Kennedy, {Deborah R.} and John Winston and Douglas Dewitt and Donald Prough",
year = "2005",
month = "4",
doi = "10.1097/00000542-200504000-00017",
language = "English (US)",
volume = "102",
pages = "806--814",
journal = "Anesthesiology",
issn = "0003-3022",
publisher = "Lippincott Williams and Wilkins",
number = "4",

}

TY - JOUR

T1 - Traumatic brain injury and hemorrhagic hypotension suppress neuroprotective gene expression in injured hippocampal neurons

AU - Hellmich, Helen

AU - Garcia, Jeanna M.

AU - Shimamura, Megumi

AU - Shah, Syed A.

AU - Avila, Marcela A.

AU - Uchida, Tatsuo

AU - Parsley, Margaret A.

AU - Capra, Bridget A.

AU - Eidson, Kristine A.

AU - Kennedy, Deborah R.

AU - Winston, John

AU - Dewitt, Douglas

AU - Prough, Donald

PY - 2005/4

Y1 - 2005/4

N2 - Background: After traumatic brain injury, memory dysfunction is due in part to damage to the hippocampus. To study the molecular mechanisms of this selective vulnerability, the authors used laser capture microdissection of neurons stained with Fluoro-Jade to directly compare gene expression in injured (Fluoro-Jade-positive) and adjacent uninjured (Fluoro-Jade-negative) rat hippocampal neurons after traumatic brain injury and traumatic brain injury plus hemorrhagic hypotension. Methods: Twelve isoflurane-anesthetized Sprague-Dawley rats underwent moderate (2.0 atm) fluid percussion traumatic brain injury followed by either normotenslon or hemorrhagic hypotension. Animals were killed 24 h after injury. Frozen brain sections were double stained with 1% cresyl violet and 0.001% Fluoro-Jade. RNA from 10 Fluoro-Jade-positive neurons and 10 Fluoro-Jade-negative neurons, obtained from the hippocampal CA1, CA3, and dentate gyrus subfields using laser capture microdissection, was linearly amplified and analyzed by quantitative ribonuclease protection assay for nine neuroprotective and apoptosis-related genes. Results: In injured CA3 neurons, expression of the neuroprotective genes glutathaione peroxidase 1, heme oxygenase 1, and brain-derived neurotrophic factor was significantly decreased compared with that of adjacent uninjured neurons. Superimposition of hemorrhagic hypotension was associated with down-regulation of neuroprotective genes in both injured and uninjured neurons of all subregions. Expression of apoptosis-related genes did not vary between injured and uninjured neurons, with or without superimposed hemorrhage. Conclusions: The authors show, in the first direct comparison of messenger RNA levels in injured and uninjured hippocampal neurons, that injured neurons express lower levels of neuroprotective genes than adjacent uninjured neurons.

AB - Background: After traumatic brain injury, memory dysfunction is due in part to damage to the hippocampus. To study the molecular mechanisms of this selective vulnerability, the authors used laser capture microdissection of neurons stained with Fluoro-Jade to directly compare gene expression in injured (Fluoro-Jade-positive) and adjacent uninjured (Fluoro-Jade-negative) rat hippocampal neurons after traumatic brain injury and traumatic brain injury plus hemorrhagic hypotension. Methods: Twelve isoflurane-anesthetized Sprague-Dawley rats underwent moderate (2.0 atm) fluid percussion traumatic brain injury followed by either normotenslon or hemorrhagic hypotension. Animals were killed 24 h after injury. Frozen brain sections were double stained with 1% cresyl violet and 0.001% Fluoro-Jade. RNA from 10 Fluoro-Jade-positive neurons and 10 Fluoro-Jade-negative neurons, obtained from the hippocampal CA1, CA3, and dentate gyrus subfields using laser capture microdissection, was linearly amplified and analyzed by quantitative ribonuclease protection assay for nine neuroprotective and apoptosis-related genes. Results: In injured CA3 neurons, expression of the neuroprotective genes glutathaione peroxidase 1, heme oxygenase 1, and brain-derived neurotrophic factor was significantly decreased compared with that of adjacent uninjured neurons. Superimposition of hemorrhagic hypotension was associated with down-regulation of neuroprotective genes in both injured and uninjured neurons of all subregions. Expression of apoptosis-related genes did not vary between injured and uninjured neurons, with or without superimposed hemorrhage. Conclusions: The authors show, in the first direct comparison of messenger RNA levels in injured and uninjured hippocampal neurons, that injured neurons express lower levels of neuroprotective genes than adjacent uninjured neurons.

UR - http://www.scopus.com/inward/record.url?scp=20144387157&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20144387157&partnerID=8YFLogxK

U2 - 10.1097/00000542-200504000-00017

DO - 10.1097/00000542-200504000-00017

M3 - Article

VL - 102

SP - 806

EP - 814

JO - Anesthesiology

JF - Anesthesiology

SN - 0003-3022

IS - 4

ER -