Effects of NF-κB oligonucleotide "decoys" on gene expression in P7 rat hippocampus after hypoxia/ischemia

Jingxin Qiu, Xiaoming Hu, Olivera Nesic, Marjorie Grafe, David K. Rassin, Thomas G. Wood, J. Regino Perez-Polo

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

"Decoy" oligonucleotides can be used as gene-specific nuclear factor (NF-κB) inhibitors to regulate gene expression. We applied two different decoy oligonucleotides that contained the NF-κB binding consensus sequences present in the immunoglobulin G (IgG)-κB and Bcl-x promoter into 7-day-old (P7) rat lateral ventricles before hypoxia/ischemia (HI) and compared their effects on gene expression in hippocampi to saline-treated, scrambled decoy-treated, or untreated hippocampi exposed to HI. Left hippocampi were collected at 12 hr after HI. Electrophoretic mobility shift assays (EMSAs) showed that the two decoy treatments had different effects on NF-κB binding to the IgG-κB and Bcl-x promoter-specific consensus sequences, respectively. We assessed the decoys' effects on gene expression 12 hr after HI using ribonuclease protection assays (RPAs) and Affymetrix DNA microarrays. RPAs showed that both decoys significantly decreased interleukin (IL)-1α mRNA levels but had no impact on IL-1β, IL-6, and IL-10 mRNA levels. IgG-κB decoys significantly decreased tumor necrosis factor (TNF)-α and TNF-β mRNA levels compared to minimal changes after treatment with Bcl-x decoys. DNA microarray analyses showed that Bcl-xL decoy treatment significantly decreased Bcl-xL mRNA levels. The decreased Bcl-xL mRNA levels after Bcl-x decoy treatment was confirmed by RPA analysis. DNA microarray data also indicated that several other genes were affected by both decoys. Our results suggest that different NF-κB decoy treatments could differentially regulate transcriptional responses to central nervous system trauma. Careful design of decoy sequences, however, is essential to acquire selective effects on cell death outcome.

Original languageEnglish (US)
Pages (from-to)108-118
Number of pages11
JournalJournal of Neuroscience Research
Volume77
Issue number1
DOIs
StatePublished - Jul 1 2004
Externally publishedYes

Fingerprint

Oligonucleotides
Hippocampus
Ischemia
Gene Expression
Ribonucleases
Messenger RNA
Oligonucleotide Array Sequence Analysis
Immunoglobulin G
Consensus Sequence
Interleukin-1
Tumor Necrosis Factor-alpha
Nervous System Trauma
Lateral Ventricles
Electrophoretic Mobility Shift Assay
Microarray Analysis
Interleukin-10
Genes
Interleukin-6
Cell Death
Central Nervous System

Keywords

  • Bcl-x, inflammation
  • Decoy
  • Hypoxia/ischemia
  • NF-κB

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Effects of NF-κB oligonucleotide "decoys" on gene expression in P7 rat hippocampus after hypoxia/ischemia. / Qiu, Jingxin; Hu, Xiaoming; Nesic, Olivera; Grafe, Marjorie; Rassin, David K.; Wood, Thomas G.; Perez-Polo, J. Regino.

In: Journal of Neuroscience Research, Vol. 77, No. 1, 01.07.2004, p. 108-118.

Research output: Contribution to journalArticle

Qiu, Jingxin ; Hu, Xiaoming ; Nesic, Olivera ; Grafe, Marjorie ; Rassin, David K. ; Wood, Thomas G. ; Perez-Polo, J. Regino. / Effects of NF-κB oligonucleotide "decoys" on gene expression in P7 rat hippocampus after hypoxia/ischemia. In: Journal of Neuroscience Research. 2004 ; Vol. 77, No. 1. pp. 108-118.
@article{3beb4927167d41dbabb89e39e0a91c70,
title = "Effects of NF-κB oligonucleotide {"}decoys{"} on gene expression in P7 rat hippocampus after hypoxia/ischemia",
abstract = "{"}Decoy{"} oligonucleotides can be used as gene-specific nuclear factor (NF-κB) inhibitors to regulate gene expression. We applied two different decoy oligonucleotides that contained the NF-κB binding consensus sequences present in the immunoglobulin G (IgG)-κB and Bcl-x promoter into 7-day-old (P7) rat lateral ventricles before hypoxia/ischemia (HI) and compared their effects on gene expression in hippocampi to saline-treated, scrambled decoy-treated, or untreated hippocampi exposed to HI. Left hippocampi were collected at 12 hr after HI. Electrophoretic mobility shift assays (EMSAs) showed that the two decoy treatments had different effects on NF-κB binding to the IgG-κB and Bcl-x promoter-specific consensus sequences, respectively. We assessed the decoys' effects on gene expression 12 hr after HI using ribonuclease protection assays (RPAs) and Affymetrix DNA microarrays. RPAs showed that both decoys significantly decreased interleukin (IL)-1α mRNA levels but had no impact on IL-1β, IL-6, and IL-10 mRNA levels. IgG-κB decoys significantly decreased tumor necrosis factor (TNF)-α and TNF-β mRNA levels compared to minimal changes after treatment with Bcl-x decoys. DNA microarray analyses showed that Bcl-xL decoy treatment significantly decreased Bcl-xL mRNA levels. The decreased Bcl-xL mRNA levels after Bcl-x decoy treatment was confirmed by RPA analysis. DNA microarray data also indicated that several other genes were affected by both decoys. Our results suggest that different NF-κB decoy treatments could differentially regulate transcriptional responses to central nervous system trauma. Careful design of decoy sequences, however, is essential to acquire selective effects on cell death outcome.",
keywords = "Bcl-x, inflammation, Decoy, Hypoxia/ischemia, NF-κB",
author = "Jingxin Qiu and Xiaoming Hu and Olivera Nesic and Marjorie Grafe and Rassin, {David K.} and Wood, {Thomas G.} and Perez-Polo, {J. Regino}",
year = "2004",
month = "7",
day = "1",
doi = "10.1002/jnr.20156",
language = "English (US)",
volume = "77",
pages = "108--118",
journal = "Journal of Neuroscience Research",
issn = "0360-4012",
publisher = "Wiley-Liss Inc.",
number = "1",

}

TY - JOUR

T1 - Effects of NF-κB oligonucleotide "decoys" on gene expression in P7 rat hippocampus after hypoxia/ischemia

AU - Qiu, Jingxin

AU - Hu, Xiaoming

AU - Nesic, Olivera

AU - Grafe, Marjorie

AU - Rassin, David K.

AU - Wood, Thomas G.

AU - Perez-Polo, J. Regino

PY - 2004/7/1

Y1 - 2004/7/1

N2 - "Decoy" oligonucleotides can be used as gene-specific nuclear factor (NF-κB) inhibitors to regulate gene expression. We applied two different decoy oligonucleotides that contained the NF-κB binding consensus sequences present in the immunoglobulin G (IgG)-κB and Bcl-x promoter into 7-day-old (P7) rat lateral ventricles before hypoxia/ischemia (HI) and compared their effects on gene expression in hippocampi to saline-treated, scrambled decoy-treated, or untreated hippocampi exposed to HI. Left hippocampi were collected at 12 hr after HI. Electrophoretic mobility shift assays (EMSAs) showed that the two decoy treatments had different effects on NF-κB binding to the IgG-κB and Bcl-x promoter-specific consensus sequences, respectively. We assessed the decoys' effects on gene expression 12 hr after HI using ribonuclease protection assays (RPAs) and Affymetrix DNA microarrays. RPAs showed that both decoys significantly decreased interleukin (IL)-1α mRNA levels but had no impact on IL-1β, IL-6, and IL-10 mRNA levels. IgG-κB decoys significantly decreased tumor necrosis factor (TNF)-α and TNF-β mRNA levels compared to minimal changes after treatment with Bcl-x decoys. DNA microarray analyses showed that Bcl-xL decoy treatment significantly decreased Bcl-xL mRNA levels. The decreased Bcl-xL mRNA levels after Bcl-x decoy treatment was confirmed by RPA analysis. DNA microarray data also indicated that several other genes were affected by both decoys. Our results suggest that different NF-κB decoy treatments could differentially regulate transcriptional responses to central nervous system trauma. Careful design of decoy sequences, however, is essential to acquire selective effects on cell death outcome.

AB - "Decoy" oligonucleotides can be used as gene-specific nuclear factor (NF-κB) inhibitors to regulate gene expression. We applied two different decoy oligonucleotides that contained the NF-κB binding consensus sequences present in the immunoglobulin G (IgG)-κB and Bcl-x promoter into 7-day-old (P7) rat lateral ventricles before hypoxia/ischemia (HI) and compared their effects on gene expression in hippocampi to saline-treated, scrambled decoy-treated, or untreated hippocampi exposed to HI. Left hippocampi were collected at 12 hr after HI. Electrophoretic mobility shift assays (EMSAs) showed that the two decoy treatments had different effects on NF-κB binding to the IgG-κB and Bcl-x promoter-specific consensus sequences, respectively. We assessed the decoys' effects on gene expression 12 hr after HI using ribonuclease protection assays (RPAs) and Affymetrix DNA microarrays. RPAs showed that both decoys significantly decreased interleukin (IL)-1α mRNA levels but had no impact on IL-1β, IL-6, and IL-10 mRNA levels. IgG-κB decoys significantly decreased tumor necrosis factor (TNF)-α and TNF-β mRNA levels compared to minimal changes after treatment with Bcl-x decoys. DNA microarray analyses showed that Bcl-xL decoy treatment significantly decreased Bcl-xL mRNA levels. The decreased Bcl-xL mRNA levels after Bcl-x decoy treatment was confirmed by RPA analysis. DNA microarray data also indicated that several other genes were affected by both decoys. Our results suggest that different NF-κB decoy treatments could differentially regulate transcriptional responses to central nervous system trauma. Careful design of decoy sequences, however, is essential to acquire selective effects on cell death outcome.

KW - Bcl-x, inflammation

KW - Decoy

KW - Hypoxia/ischemia

KW - NF-κB

UR - http://www.scopus.com/inward/record.url?scp=3042806584&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3042806584&partnerID=8YFLogxK

U2 - 10.1002/jnr.20156

DO - 10.1002/jnr.20156

M3 - Article

VL - 77

SP - 108

EP - 118

JO - Journal of Neuroscience Research

JF - Journal of Neuroscience Research

SN - 0360-4012

IS - 1

ER -