Comparison of formaldehyde-preperfused frozen and freshly frozen tissue preparation for the in situ hybridization for alpha-tubulin messenger RNA in the rat brain

Masato Asanuma, N. Ogawa, K. Mizukawa, K. Haba, A. Mori

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In situ hybridization histochemistry, using the alpha-tubulin oligonucleotide probe, was performed employing two tissue freezing methods, the preperfuse-freezing method and the freshly frozen method, to evaluate methodological differences in the messenger RNA (mRNA) levels of the brain. The mRNA levels in the sections from the freshly frozen groups were lower than those in the preperfused group, particularly in the cerebral cortex, the striatum, the hippocampal CA1 and CA4 fields, and the habenular nuclei. Therefore, employing the freshly frozen method, there is a possibility that the mRNA levels in these regions may be underestimated. The hybridization signals of groups placed on an ice-bed for 5 min and 15 min were lower than those of the immediately frozen group in the cingulate, the temporal, and the retrosplenial cortex, the CA1 field, and even in the medial and the lateral thalamic nuclei where no significant difference was seen between the perfused group and the immediately frozen group of tissues. When employing the freshly frozen method, the removed brain should be frozen as fast as possible and the period from decapitation to freezing should be kept strictly constant.

Original languageEnglish
Pages (from-to)183-192
Number of pages10
JournalResearch Communications in Chemical Pathology and Pharmacology
Volume70
Issue number2
Publication statusPublished - 1990

Fingerprint

Tubulin
Freezing
Formaldehyde
In Situ Hybridization
Rats
Brain
Tissue
Messenger RNA
Oligonucleotide Probes
Ice
Habenula
Lateral Thalamic Nuclei
Mediodorsal Thalamic Nucleus
Decapitation
Dentate Gyrus
Frozen Sections
Temporal Lobe
Cerebral Cortex

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

Cite this

@article{52e4ee305a4d4c9792f4756d51e05649,
title = "Comparison of formaldehyde-preperfused frozen and freshly frozen tissue preparation for the in situ hybridization for alpha-tubulin messenger RNA in the rat brain",
abstract = "In situ hybridization histochemistry, using the alpha-tubulin oligonucleotide probe, was performed employing two tissue freezing methods, the preperfuse-freezing method and the freshly frozen method, to evaluate methodological differences in the messenger RNA (mRNA) levels of the brain. The mRNA levels in the sections from the freshly frozen groups were lower than those in the preperfused group, particularly in the cerebral cortex, the striatum, the hippocampal CA1 and CA4 fields, and the habenular nuclei. Therefore, employing the freshly frozen method, there is a possibility that the mRNA levels in these regions may be underestimated. The hybridization signals of groups placed on an ice-bed for 5 min and 15 min were lower than those of the immediately frozen group in the cingulate, the temporal, and the retrosplenial cortex, the CA1 field, and even in the medial and the lateral thalamic nuclei where no significant difference was seen between the perfused group and the immediately frozen group of tissues. When employing the freshly frozen method, the removed brain should be frozen as fast as possible and the period from decapitation to freezing should be kept strictly constant.",
author = "Masato Asanuma and N. Ogawa and K. Mizukawa and K. Haba and A. Mori",
year = "1990",
language = "English",
volume = "70",
pages = "183--192",
journal = "Research Communications in Molecular Pathology and Pharmacology",
issn = "1078-0297",
publisher = "PJD Publications Ltd",
number = "2",

}

TY - JOUR

T1 - Comparison of formaldehyde-preperfused frozen and freshly frozen tissue preparation for the in situ hybridization for alpha-tubulin messenger RNA in the rat brain

AU - Asanuma, Masato

AU - Ogawa, N.

AU - Mizukawa, K.

AU - Haba, K.

AU - Mori, A.

PY - 1990

Y1 - 1990

N2 - In situ hybridization histochemistry, using the alpha-tubulin oligonucleotide probe, was performed employing two tissue freezing methods, the preperfuse-freezing method and the freshly frozen method, to evaluate methodological differences in the messenger RNA (mRNA) levels of the brain. The mRNA levels in the sections from the freshly frozen groups were lower than those in the preperfused group, particularly in the cerebral cortex, the striatum, the hippocampal CA1 and CA4 fields, and the habenular nuclei. Therefore, employing the freshly frozen method, there is a possibility that the mRNA levels in these regions may be underestimated. The hybridization signals of groups placed on an ice-bed for 5 min and 15 min were lower than those of the immediately frozen group in the cingulate, the temporal, and the retrosplenial cortex, the CA1 field, and even in the medial and the lateral thalamic nuclei where no significant difference was seen between the perfused group and the immediately frozen group of tissues. When employing the freshly frozen method, the removed brain should be frozen as fast as possible and the period from decapitation to freezing should be kept strictly constant.

AB - In situ hybridization histochemistry, using the alpha-tubulin oligonucleotide probe, was performed employing two tissue freezing methods, the preperfuse-freezing method and the freshly frozen method, to evaluate methodological differences in the messenger RNA (mRNA) levels of the brain. The mRNA levels in the sections from the freshly frozen groups were lower than those in the preperfused group, particularly in the cerebral cortex, the striatum, the hippocampal CA1 and CA4 fields, and the habenular nuclei. Therefore, employing the freshly frozen method, there is a possibility that the mRNA levels in these regions may be underestimated. The hybridization signals of groups placed on an ice-bed for 5 min and 15 min were lower than those of the immediately frozen group in the cingulate, the temporal, and the retrosplenial cortex, the CA1 field, and even in the medial and the lateral thalamic nuclei where no significant difference was seen between the perfused group and the immediately frozen group of tissues. When employing the freshly frozen method, the removed brain should be frozen as fast as possible and the period from decapitation to freezing should be kept strictly constant.

UR - http://www.scopus.com/inward/record.url?scp=0025252490&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025252490&partnerID=8YFLogxK

M3 - Article

C2 - 2277864

AN - SCOPUS:0025252490

VL - 70

SP - 183

EP - 192

JO - Research Communications in Molecular Pathology and Pharmacology

JF - Research Communications in Molecular Pathology and Pharmacology

SN - 1078-0297

IS - 2

ER -