A method for the histochemical localization of digestive enzymes in whole freeze‐substituted larval fish embedded in glycol methacrylate
Identifieur interne : 000A07 ( Main/Merge ); précédent : 000A06; suivant : 000A08A method for the histochemical localization of digestive enzymes in whole freeze‐substituted larval fish embedded in glycol methacrylate
Auteurs : Grant W. Vandenberg [Canada] ; Diane Gagnon [Canada] ; Shannon L. Scott [Canada] ; Joël De Lanoüe [Canada]Source :
- Aquaculture Research [ 1355-557X ] ; 2009-04.
English descriptors
- KwdEn :
Abstract
In order to study the localization of digestive enzymes in larval walleye (Sander vitreus vitreus), a novel method of low‐temperature processing of whole, unfixed larvae and subsequent embedding in glycol methacrylate (GMA) was developed. Larval walleye aged 2–10 days post hatch were flash‐frozen in liquid nitrogen and transferred into pre‐chilled acetone for 12 h at a temperature of −25 °C. Acetone was gradually replaced with increasing concentrations of GMA resin monomer over a 6‐h period. The polymer (100%) was further allowed to infiltrate the larvae for 36 h. Resin‐embedded larvae were transferred to embedding moulds and polymerized overnight at −25 °C. Four micrometre sections were stained to identify either alkaline phosphatase, non‐specific esterase, aminopeptidase M or dipeptidyl peptidase IV. All enzymes studied were readily detected and accurately localized, and no enzyme diffusion was observed. Therefore, freeze substitution combined with low‐temperature GMA embedding allows the maintenance of excellent tissue morphology and accurate enzyme localization in whole larval walleye specimens from 2 to 10 days post hatch. It is recommended, however, that samples be frozen in pre‐cooled fluorocarbon‐based liquid coolants in order to assure optimal tissue preservation.
Url:
DOI: 10.1111/j.1365-2109.2009.02168.x
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 000821
- to stream Istex, to step Curation: 000819
- to stream Istex, to step Checkpoint: 000468
Links to Exploration step
ISTEX:69CE395D78143037094B20FADBD8302DAC469DFELe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">A method for the histochemical localization of digestive enzymes in whole freeze‐substituted larval fish embedded in glycol methacrylate</title><author><name sortKey="Vandenberg, Grant W" sort="Vandenberg, Grant W" uniqKey="Vandenberg G" first="Grant W" last="Vandenberg">Grant W. Vandenberg</name></author><author><name sortKey="Gagnon, Diane" sort="Gagnon, Diane" uniqKey="Gagnon D" first="Diane" last="Gagnon">Diane Gagnon</name></author><author><name sortKey="Scott, Shannon L" sort="Scott, Shannon L" uniqKey="Scott S" first="Shannon L" last="Scott">Shannon L. Scott</name></author><author><name sortKey="De Lanoue, Joel" sort="De Lanoue, Joel" uniqKey="De Lanoue J" first="Joël" last="De Lanoüe">Joël De Lanoüe</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:69CE395D78143037094B20FADBD8302DAC469DFE</idno><date when="2009" year="2009">2009</date><idno type="doi">10.1111/j.1365-2109.2009.02168.x</idno><idno type="url">https://api.istex.fr/document/69CE395D78143037094B20FADBD8302DAC469DFE/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">000821</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000821</idno><idno type="wicri:Area/Istex/Curation">000819</idno><idno type="wicri:Area/Istex/Checkpoint">000468</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000468</idno><idno type="wicri:doubleKey">1355-557X:2009:Vandenberg G:a:method:for</idno><idno type="wicri:Area/Main/Merge">000A07</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">A method for the histochemical localization of digestive enzymes in whole freeze‐substituted larval fish embedded in glycol methacrylate</title><author><name sortKey="Vandenberg, Grant W" sort="Vandenberg, Grant W" uniqKey="Vandenberg G" first="Grant W" last="Vandenberg">Grant W. Vandenberg</name><affiliation wicri:level="1"><country>Canada</country><wicri:regionArea>Groupe de recherche en recyclage biologique et aquiculture. Département des sciences animales, Université Laval, QC</wicri:regionArea><wicri:noRegion>QC</wicri:noRegion></affiliation></author><author><name sortKey="Gagnon, Diane" sort="Gagnon, Diane" uniqKey="Gagnon D" first="Diane" last="Gagnon">Diane Gagnon</name><affiliation wicri:level="1"><country>Canada</country><wicri:regionArea>Groupe de recherche en recyclage biologique et aquiculture. Département des sciences animales, Université Laval, QC</wicri:regionArea><wicri:noRegion>QC</wicri:noRegion></affiliation></author><author><name sortKey="Scott, Shannon L" sort="Scott, Shannon L" uniqKey="Scott S" first="Shannon L" last="Scott">Shannon L. Scott</name><affiliation wicri:level="1"><country>Canada</country><wicri:regionArea>Groupe de recherche en recyclage biologique et aquiculture. Département des sciences animales, Université Laval, QC</wicri:regionArea><wicri:noRegion>QC</wicri:noRegion></affiliation></author><author><name sortKey="De Lanoue, Joel" sort="De Lanoue, Joel" uniqKey="De Lanoue J" first="Joël" last="De Lanoüe">Joël De Lanoüe</name><affiliation wicri:level="1"><country>Canada</country><wicri:regionArea>Groupe de recherche en recyclage biologique et aquiculture. Département des sciences animales, Université Laval, QC</wicri:regionArea><wicri:noRegion>QC</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Aquaculture Research</title><idno type="ISSN">1355-557X</idno><idno type="eISSN">1365-2109</idno><imprint><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2009-04">2009-04</date><biblScope unit="volume">40</biblScope><biblScope unit="issue">7</biblScope><biblScope unit="page" from="818">818</biblScope><biblScope unit="page" to="824">824</biblScope></imprint><idno type="ISSN">1355-557X</idno></series><idno type="istex">69CE395D78143037094B20FADBD8302DAC469DFE</idno><idno type="DOI">10.1111/j.1365-2109.2009.02168.x</idno><idno type="ArticleID">ARE2168</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1355-557X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>enzyme</term><term>fish larvae</term><term>freeze substitution</term><term>histochemistry</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In order to study the localization of digestive enzymes in larval walleye (Sander vitreus vitreus), a novel method of low‐temperature processing of whole, unfixed larvae and subsequent embedding in glycol methacrylate (GMA) was developed. Larval walleye aged 2–10 days post hatch were flash‐frozen in liquid nitrogen and transferred into pre‐chilled acetone for 12 h at a temperature of −25 °C. Acetone was gradually replaced with increasing concentrations of GMA resin monomer over a 6‐h period. The polymer (100%) was further allowed to infiltrate the larvae for 36 h. Resin‐embedded larvae were transferred to embedding moulds and polymerized overnight at −25 °C. Four micrometre sections were stained to identify either alkaline phosphatase, non‐specific esterase, aminopeptidase M or dipeptidyl peptidase IV. All enzymes studied were readily detected and accurately localized, and no enzyme diffusion was observed. Therefore, freeze substitution combined with low‐temperature GMA embedding allows the maintenance of excellent tissue morphology and accurate enzyme localization in whole larval walleye specimens from 2 to 10 days post hatch. It is recommended, however, that samples be frozen in pre‐cooled fluorocarbon‐based liquid coolants in order to assure optimal tissue preservation.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Eau/explor/EsturgeonV1/Data/Main/Merge
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000A07 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Merge/biblio.hfd -nk 000A07 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Eau
|area= EsturgeonV1
|flux= Main
|étape= Merge
|type= RBID
|clé= ISTEX:69CE395D78143037094B20FADBD8302DAC469DFE
|texte= A method for the histochemical localization of digestive enzymes in whole freeze‐substituted larval fish embedded in glycol methacrylate
}}
| This area was generated with Dilib version V0.6.27. |  |