Detection of glyceraldehyde 3‐phosphate dehydrogenase messenger RNA using a peptide nucleic acid probe in paraffin‐embedded archival specimens
Identifieur interne : 003167 ( Main/Exploration ); précédent : 003166; suivant : 003168Detection of glyceraldehyde 3‐phosphate dehydrogenase messenger RNA using a peptide nucleic acid probe in paraffin‐embedded archival specimens
Auteurs : Makoto Hiroyasu ; Shinya Akatsuka ; Tomoyuki Shirase ; Yoshinobu Toda [Japon] ; Hiroshi Hiai ; Shinya Toyokuni [Japon]Source :
- Pathology International [ 1320-5463 ] ; 2004-04.
English descriptors
- Teeft :
- Acid probe, Adenocarcinoma, Archival, Archival autopsy specimens, Autopsy, Autopsy specimens, Carcinoma, Cell carcinoma, Classic riboprobes, Degeneration, Dehydrogenase, Eosin staining, Formalin, Gapdh, Gapdh mrna, Genome, Glyceraldehyde, Glyceraldehyde dehydrogenase, Glyceraldehyde dehydrogenase messenger, Graduate school, High cost, Human genome, Human genome project, Hybridization, Inverse correlation, Kyoto university, Liver specimens, Mild change, Moderate change, Mrna, Mrna analysis, Myelogenous leukemia, Normal limit, Oligonucleotide, Peptide, Polymerase chain reaction, Postmortal degeneration, Premortal, Present experiments, Room temperature, Severe change, Small fragments, Staining intensity, Tissue degeneration, Tissue specimens, Toyokuni, Typical examples, Xation.
Abstract
Although the human genome project has been completed, the functions of many genes remain undetermined. In situ hybridization (ISH) is a key method for identifying cells in which a given messenger RNA is transcribed. Paraffin‐embedded specimens remain precious materials for research, but preservation of high‐quality RNA in these specimens is not expected unless ample caution was taken during fixation. Peptide nucleic acid (PNA) is a recently developed hybrid molecule with genetic information that has high stability and high affinity to the complementary DNA or RNA. We applied a PNA probe to mRNA ISH of liver specimens obtained by autopsy and embedded in paraffin 28–48 years ago. An 18‐mer PNA probe for glyceraldehyde 3‐phosphate dehydrogenase was used. Staining was then analyzed in association with morphology by hematoxylin and eosin staining, and with the time between death of the patient and tissue fixation. Notably, specimens fixed with formalin and embedded in paraffin 48 years ago yielded excellent results if the time before fixation was short enough (<8 h). There was a significant inverse correlation between the intensity of ISH staining and the time before fixation. Oligonucleotide PNA probe, albeit at high cost, would increase the value of paraffin‐embedded specimens in storage for use in human medical research.
Url:
DOI: 10.1111/j.1440-1827.2004.01620.x
Affiliations:
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Shirase</name><affiliation><wicri:noCountry code="no comma">Department of Pathology and Biology of Diseases and</wicri:noCountry></affiliation></author><author><name sortKey="Toda, Yoshinobu" sort="Toda, Yoshinobu" uniqKey="Toda Y" first="Yoshinobu" last="Toda">Yoshinobu Toda</name><affiliation wicri:level="4"><country xml:lang="fr">Japon</country><wicri:regionArea>Center for Anatomical Studies, Graduate School of Medicine, Kyoto University, Kyoto</wicri:regionArea><orgName type="university">Université de Kyoto</orgName><placeName><settlement type="city">Kyoto</settlement><region type="prefecture">Région du Kansai</region></placeName></affiliation></author><author><name sortKey="Hiai, Hiroshi" sort="Hiai, Hiroshi" uniqKey="Hiai H" first="Hiroshi" last="Hiai">Hiroshi Hiai</name><affiliation><wicri:noCountry code="no comma">Department of Pathology and Biology of Diseases and</wicri:noCountry></affiliation></author><author><name sortKey="Toyokuni, Shinya" sort="Toyokuni, Shinya" uniqKey="Toyokuni S" first="Shinya" last="Toyokuni">Shinya Toyokuni</name><affiliation></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Japon</country></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Pathology International</title><title level="j" type="alt">PATHOLOGY INTERNATIONAL</title><idno type="ISSN">1320-5463</idno><idno type="eISSN">1440-1827</idno><imprint><biblScope unit="vol">54</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="251">251</biblScope><biblScope unit="page" to="255">255</biblScope><biblScope unit="page-count">5</biblScope><publisher>Blackwell Science Pty</publisher><pubPlace>Melbourne, Australia</pubPlace><date type="published" when="2004-04">2004-04</date></imprint><idno type="ISSN">1320-5463</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1320-5463</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acid probe</term><term>Adenocarcinoma</term><term>Archival</term><term>Archival autopsy specimens</term><term>Autopsy</term><term>Autopsy specimens</term><term>Carcinoma</term><term>Cell carcinoma</term><term>Classic riboprobes</term><term>Degeneration</term><term>Dehydrogenase</term><term>Eosin staining</term><term>Formalin</term><term>Gapdh</term><term>Gapdh mrna</term><term>Genome</term><term>Glyceraldehyde</term><term>Glyceraldehyde dehydrogenase</term><term>Glyceraldehyde dehydrogenase messenger</term><term>Graduate school</term><term>High cost</term><term>Human genome</term><term>Human genome project</term><term>Hybridization</term><term>Inverse correlation</term><term>Kyoto university</term><term>Liver specimens</term><term>Mild change</term><term>Moderate change</term><term>Mrna</term><term>Mrna analysis</term><term>Myelogenous leukemia</term><term>Normal limit</term><term>Oligonucleotide</term><term>Peptide</term><term>Polymerase chain reaction</term><term>Postmortal degeneration</term><term>Premortal</term><term>Present experiments</term><term>Room temperature</term><term>Severe change</term><term>Small fragments</term><term>Staining intensity</term><term>Tissue degeneration</term><term>Tissue specimens</term><term>Toyokuni</term><term>Typical examples</term><term>Xation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Although the human genome project has been completed, the functions of many genes remain undetermined. In situ hybridization (ISH) is a key method for identifying cells in which a given messenger RNA is transcribed. Paraffin‐embedded specimens remain precious materials for research, but preservation of high‐quality RNA in these specimens is not expected unless ample caution was taken during fixation. Peptide nucleic acid (PNA) is a recently developed hybrid molecule with genetic information that has high stability and high affinity to the complementary DNA or RNA. We applied a PNA probe to mRNA ISH of liver specimens obtained by autopsy and embedded in paraffin 28–48 years ago. An 18‐mer PNA probe for glyceraldehyde 3‐phosphate dehydrogenase was used. Staining was then analyzed in association with morphology by hematoxylin and eosin staining, and with the time between death of the patient and tissue fixation. Notably, specimens fixed with formalin and embedded in paraffin 48 years ago yielded excellent results if the time before fixation was short enough (<8 h). There was a significant inverse correlation between the intensity of ISH staining and the time before fixation. Oligonucleotide PNA probe, albeit at high cost, would increase the value of paraffin‐embedded specimens in storage for use in human medical research.</div></front></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région du Kansai</li></region><settlement><li>Kyoto</li></settlement><orgName><li>Université de Kyoto</li></orgName></list><tree><noCountry><name sortKey="Akatsuka, Shinya" sort="Akatsuka, Shinya" uniqKey="Akatsuka S" first="Shinya" last="Akatsuka">Shinya Akatsuka</name><name sortKey="Hiai, Hiroshi" sort="Hiai, Hiroshi" uniqKey="Hiai H" first="Hiroshi" last="Hiai">Hiroshi Hiai</name><name sortKey="Hiroyasu, Makoto" sort="Hiroyasu, Makoto" uniqKey="Hiroyasu M" first="Makoto" last="Hiroyasu">Makoto Hiroyasu</name><name sortKey="Shirase, Tomoyuki" sort="Shirase, Tomoyuki" uniqKey="Shirase T" first="Tomoyuki" last="Shirase">Tomoyuki Shirase</name></noCountry><country name="Japon"><region name="Région du Kansai"><name sortKey="Toda, Yoshinobu" sort="Toda, Yoshinobu" uniqKey="Toda Y" first="Yoshinobu" last="Toda">Yoshinobu Toda</name></region><name sortKey="Toyokuni, Shinya" sort="Toyokuni, Shinya" uniqKey="Toyokuni S" first="Shinya" last="Toyokuni">Shinya Toyokuni</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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