c-fos gene expression is increased in the paraventricular hypothalamic nucleus of Sprague-Dawley rats with visceral pain induced by acetic acid without detectable changes of corticotrophin-releasing factor mRNA: A quantitative approach with an image analysis system

Bang H. Hwang, Hung Ming Chang, Zhu H. Gu, Ryoji Suzuki

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

This study is the first of its kind to demonstrate that c-Fos immunoreactivity (ir) together with c-fos mRNA in their immediately adjacent tissue sections of a discrete brain region can be reliably measured. The c-fos gene expression in the paraventricular hypothalamic nucleus (PVN) of Sprague-Dawley rats for an animal model for visceral or somatovisceral pain induced by 2% acetic acid (AA) was used in this study. Specifically, c-fos mRNA signals were measured by quantitative autoradiography after in situ hybridization using c-fos ohgodeoxynucleotide probe, and c-Fos-ir signals were represented by c-Fos immunostaining, as detected using c-Fos antibody in a regular immunohistochemistry. Signals from both c-Fos-ir and c-fos mRNA in the PVN were measured from their immediately adjacent cryostat sections. For the measurement of c-Fos-ir, it was carried out by reading 10 rectangles (1,000 μm2/rectangle) on each PVN section with c-Fos immunostaining. Specific signals were obtained from subtracting the nonspecific background signal from the total signals using a computer-assisted image analysis system. Results indicated that the AA treatment induced a significant increase of both c-Fos-ir and c-fos mRNA in the PVN. Interestingly, there was no increase of corticotrophin-releasing factor (CRF) mRNA expression in the PVN and central nucleus of the amygdala of Sprague-Dawley rats subjected to the AA treatment. In summary, this study has demonstrated that c-Fos-ir in the PVN with an anatomical resolution can be semiquantitatively measured after immunohistochemistry using an image analysis system, and that increased c-fos mRNA in the PVN 1 hr after the AA treatment is associated with no changes of the CRF mRNA expression.

Original languageEnglish
Pages (from-to)406-413
Number of pages8
JournalAnatomical Record
Volume290
Issue number4
DOIs
Publication statusPublished - Apr 2007
Externally publishedYes

Fingerprint

paraventricular hypothalamic nucleus
Visceral Pain
fos Genes
corticotropin-releasing hormone
Paraventricular Hypothalamic Nucleus
Corticotropin-Releasing Hormone
Acetic Acid
image analysis
acetic acid
gene expression
Sprague Dawley Rats
pain
Gene Expression
Messenger RNA
rats
immunohistochemistry
animal models
Immunohistochemistry
amygdala
Computer-Assisted Image Processing

Keywords

  • c-Fos immunostaining
  • c-fos mRNA
  • Paraventricular hypothalamic nucleus
  • Visceral pain

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Biotechnology
  • Anatomy
  • Histology

Cite this

@article{33b9cd41a52f42d39a998e4d26a4a82a,
title = "c-fos gene expression is increased in the paraventricular hypothalamic nucleus of Sprague-Dawley rats with visceral pain induced by acetic acid without detectable changes of corticotrophin-releasing factor mRNA: A quantitative approach with an image analysis system",
abstract = "This study is the first of its kind to demonstrate that c-Fos immunoreactivity (ir) together with c-fos mRNA in their immediately adjacent tissue sections of a discrete brain region can be reliably measured. The c-fos gene expression in the paraventricular hypothalamic nucleus (PVN) of Sprague-Dawley rats for an animal model for visceral or somatovisceral pain induced by 2{\%} acetic acid (AA) was used in this study. Specifically, c-fos mRNA signals were measured by quantitative autoradiography after in situ hybridization using c-fos ohgodeoxynucleotide probe, and c-Fos-ir signals were represented by c-Fos immunostaining, as detected using c-Fos antibody in a regular immunohistochemistry. Signals from both c-Fos-ir and c-fos mRNA in the PVN were measured from their immediately adjacent cryostat sections. For the measurement of c-Fos-ir, it was carried out by reading 10 rectangles (1,000 μm2/rectangle) on each PVN section with c-Fos immunostaining. Specific signals were obtained from subtracting the nonspecific background signal from the total signals using a computer-assisted image analysis system. Results indicated that the AA treatment induced a significant increase of both c-Fos-ir and c-fos mRNA in the PVN. Interestingly, there was no increase of corticotrophin-releasing factor (CRF) mRNA expression in the PVN and central nucleus of the amygdala of Sprague-Dawley rats subjected to the AA treatment. In summary, this study has demonstrated that c-Fos-ir in the PVN with an anatomical resolution can be semiquantitatively measured after immunohistochemistry using an image analysis system, and that increased c-fos mRNA in the PVN 1 hr after the AA treatment is associated with no changes of the CRF mRNA expression.",
keywords = "c-Fos immunostaining, c-fos mRNA, Paraventricular hypothalamic nucleus, Visceral pain",
author = "Hwang, {Bang H.} and Chang, {Hung Ming} and Gu, {Zhu H.} and Ryoji Suzuki",
year = "2007",
month = "4",
doi = "10.1002/ar.20495",
language = "English",
volume = "290",
pages = "406--413",
journal = "Anatomical Record",
issn = "0003-276X",
publisher = "John Wiley and Sons Inc.",
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TY - JOUR

T1 - c-fos gene expression is increased in the paraventricular hypothalamic nucleus of Sprague-Dawley rats with visceral pain induced by acetic acid without detectable changes of corticotrophin-releasing factor mRNA

T2 - A quantitative approach with an image analysis system

AU - Hwang, Bang H.

AU - Chang, Hung Ming

AU - Gu, Zhu H.

AU - Suzuki, Ryoji

PY - 2007/4

Y1 - 2007/4

N2 - This study is the first of its kind to demonstrate that c-Fos immunoreactivity (ir) together with c-fos mRNA in their immediately adjacent tissue sections of a discrete brain region can be reliably measured. The c-fos gene expression in the paraventricular hypothalamic nucleus (PVN) of Sprague-Dawley rats for an animal model for visceral or somatovisceral pain induced by 2% acetic acid (AA) was used in this study. Specifically, c-fos mRNA signals were measured by quantitative autoradiography after in situ hybridization using c-fos ohgodeoxynucleotide probe, and c-Fos-ir signals were represented by c-Fos immunostaining, as detected using c-Fos antibody in a regular immunohistochemistry. Signals from both c-Fos-ir and c-fos mRNA in the PVN were measured from their immediately adjacent cryostat sections. For the measurement of c-Fos-ir, it was carried out by reading 10 rectangles (1,000 μm2/rectangle) on each PVN section with c-Fos immunostaining. Specific signals were obtained from subtracting the nonspecific background signal from the total signals using a computer-assisted image analysis system. Results indicated that the AA treatment induced a significant increase of both c-Fos-ir and c-fos mRNA in the PVN. Interestingly, there was no increase of corticotrophin-releasing factor (CRF) mRNA expression in the PVN and central nucleus of the amygdala of Sprague-Dawley rats subjected to the AA treatment. In summary, this study has demonstrated that c-Fos-ir in the PVN with an anatomical resolution can be semiquantitatively measured after immunohistochemistry using an image analysis system, and that increased c-fos mRNA in the PVN 1 hr after the AA treatment is associated with no changes of the CRF mRNA expression.

AB - This study is the first of its kind to demonstrate that c-Fos immunoreactivity (ir) together with c-fos mRNA in their immediately adjacent tissue sections of a discrete brain region can be reliably measured. The c-fos gene expression in the paraventricular hypothalamic nucleus (PVN) of Sprague-Dawley rats for an animal model for visceral or somatovisceral pain induced by 2% acetic acid (AA) was used in this study. Specifically, c-fos mRNA signals were measured by quantitative autoradiography after in situ hybridization using c-fos ohgodeoxynucleotide probe, and c-Fos-ir signals were represented by c-Fos immunostaining, as detected using c-Fos antibody in a regular immunohistochemistry. Signals from both c-Fos-ir and c-fos mRNA in the PVN were measured from their immediately adjacent cryostat sections. For the measurement of c-Fos-ir, it was carried out by reading 10 rectangles (1,000 μm2/rectangle) on each PVN section with c-Fos immunostaining. Specific signals were obtained from subtracting the nonspecific background signal from the total signals using a computer-assisted image analysis system. Results indicated that the AA treatment induced a significant increase of both c-Fos-ir and c-fos mRNA in the PVN. Interestingly, there was no increase of corticotrophin-releasing factor (CRF) mRNA expression in the PVN and central nucleus of the amygdala of Sprague-Dawley rats subjected to the AA treatment. In summary, this study has demonstrated that c-Fos-ir in the PVN with an anatomical resolution can be semiquantitatively measured after immunohistochemistry using an image analysis system, and that increased c-fos mRNA in the PVN 1 hr after the AA treatment is associated with no changes of the CRF mRNA expression.

KW - c-Fos immunostaining

KW - c-fos mRNA

KW - Paraventricular hypothalamic nucleus

KW - Visceral pain

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