Staining of sturgeon spermatozoa with trypsin inhibitor from soybean, Alexa Fluor® 488 conjugate for visualization of sturgeon acrosome
Identifieur interne : 000238 ( Istex/Corpus ); précédent : 000237; suivant : 000239Staining of sturgeon spermatozoa with trypsin inhibitor from soybean, Alexa Fluor® 488 conjugate for visualization of sturgeon acrosome
Auteurs : M. Psenicka ; J. Cosson ; S. M. H. Alavi ; M. Rodina ; V. Kaspar ; D. Gela ; O. Linhart ; A. CiereszkoSource :
- Journal of Applied Ichthyology [ 0175-8659 ] ; 2008-08.
Abstract
Trypsin‐like activity, similar to acrosin, is present in sturgeon spermatozoa and can be a potential target for trypsin inhibitors. The objective of this work was to use a fluorescent soybean trypsin inhibitor (SBTI) conjugate with the Alexa Fluor® 488 dye for visualization of the sturgeon acrosome. After incubation with SBTI‐Alexa, a strong signal was observed both in the acrosome and midpiece or implantation fossa region. We have also found that SBTI‐Alexa staining can be combined with PI viability test. Detailed examination of staining pattern revealed that SBTI‐Alexa can stain either acrosome or whole sperm. Staining of whole sperm correlated with dead staining (r2 = 0.94, P < 0.01). However, in fresh semen most cells (93–97%) were not stained with SBTI‐Alexa, probably due to intact acrosomes. Further studies should test if SBTI‐Alexa can be applied to monitor the acrosome status during the acrosome reaction and cryopreservation.
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DOI: 10.1111/j.1439-0426.2008.01140.x
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ISTEX:D410D188F9BE140E2E47C8262B3AE00BE1D95A10Le document en format XML
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Staining of whole sperm correlated with dead staining (r2 = 0.94, P > 0.01). However, in fresh semen most cells (93–97%) were not stained with SBTI‐Alexa, probably due to intact acrosomes. 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The objective of this work was to use a fluorescent soybean trypsin inhibitor (SBTI) conjugate with the Alexa Fluor® 488 dye for visualization of the sturgeon acrosome. After incubation with SBTI‐Alexa, a strong signal was observed both in the acrosome and midpiece or implantation fossa region. We have also found that SBTI‐Alexa staining can be combined with PI viability test. Detailed examination of staining pattern revealed that SBTI‐Alexa can stain either acrosome or whole sperm. Staining of whole sperm correlated with dead staining (r2 = 0.94, P < 0.01). However, in fresh semen most cells (93–97%) were not stained with SBTI‐Alexa, probably due to intact acrosomes. 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E‐mail: <email>acieresz@pan.olsztyn.pl</email></correspondenceTo><linkGroup><link type="toTypesetVersion" href="file:JAI.JAI1140.pdf"></link></linkGroup></publicationMeta><contentMeta><unparsedEditorialHistory>Received: September 15, 2007 Accepted: March 12, 2008</unparsedEditorialHistory><countGroup><count type="figureTotal" number="3"></count><count type="tableTotal" number="0"></count></countGroup><titleGroup><title type="main">Staining of sturgeon spermatozoa with trypsin inhibitor from soybean, Alexa Fluor<sup>®</sup> 488 conjugate for visualization of sturgeon acrosome</title><title type="shortAuthors">M. Psenicka et al.</title><title type="short">SBTI‐Alexa staining of sturgeon acrosome</title></titleGroup><creators><creator creatorRole="author" xml:id="cr1" affiliationRef="#a1"><personName><givenNames>M.</givenNames><familyName>Psenicka</familyName></personName></creator><creator creatorRole="author" xml:id="cr2" affiliationRef="#a2"><personName><givenNames>J.</givenNames><familyName>Cosson</familyName></personName></creator><creator creatorRole="author" xml:id="cr3" affiliationRef="#a1"><personName><givenNames>S. M. H.</givenNames><familyName>Alavi</familyName></personName></creator><creator creatorRole="author" xml:id="cr4" affiliationRef="#a1"><personName><givenNames>M.</givenNames><familyName>Rodina</familyName></personName></creator><creator creatorRole="author" xml:id="cr5" affiliationRef="#a1"><personName><givenNames>V.</givenNames><familyName>Kaspar</familyName></personName></creator><creator creatorRole="author" xml:id="cr6" affiliationRef="#a1"><personName><givenNames>D.</givenNames><familyName>Gela</familyName></personName></creator><creator creatorRole="author" xml:id="cr7" affiliationRef="#a1"><personName><givenNames>O.</givenNames><familyName>Linhart</familyName></personName></creator><creator creatorRole="author" xml:id="cr8" affiliationRef="#a3"><personName><givenNames>A.</givenNames><familyName>Ciereszko</familyName></personName></creator></creators><affiliationGroup><affiliation xml:id="a1" countryCode="CZ"><unparsedAffiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</unparsedAffiliation></affiliation><affiliation xml:id="a2" countryCode="FR"><unparsedAffiliation>UMR 7009 CNRS – Univ. Paris VI, Observatore de Villefranche/mer, Marine Station, Villefranche‐sur‐mer, France</unparsedAffiliation></affiliation><affiliation xml:id="a3" countryCode="PL"><unparsedAffiliation>Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland</unparsedAffiliation></affiliation></affiliationGroup><abstractGroup><abstract type="main" xml:lang="en"><title type="main">Summary</title><p>Trypsin‐like activity, similar to acrosin, is present in sturgeon spermatozoa and can be a potential target for trypsin inhibitors. The objective of this work was to use a fluorescent soybean trypsin inhibitor (SBTI) conjugate with the Alexa Fluor<sup>®</sup> 488 dye for visualization of the sturgeon acrosome. After incubation with SBTI‐Alexa, a strong signal was observed both in the acrosome and midpiece or implantation fossa region. We have also found that SBTI‐Alexa staining can be combined with PI viability test. Detailed examination of staining pattern revealed that SBTI‐Alexa can stain either acrosome or whole sperm. Staining of whole sperm correlated with dead staining (<i>r</i><sup>2</sup> = 0.94, P < 0.01). However, in fresh semen most cells (93–97%) were not stained with SBTI‐Alexa, probably due to intact acrosomes. Further studies should test if SBTI‐Alexa can be applied to monitor the acrosome status during the acrosome reaction and cryopreservation.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Staining of sturgeon spermatozoa with trypsin inhibitor from soybean, Alexa Fluor® 488 conjugate for visualization of sturgeon acrosome</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>SBTI‐Alexa staining of sturgeon acrosome</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Staining of sturgeon spermatozoa with trypsin inhibitor from soybean, Alexa Fluor® 488 conjugate for visualization of sturgeon acrosome</title></titleInfo><name type="personal"><namePart type="given">M.</namePart><namePart type="family">Psenicka</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J.</namePart><namePart type="family">Cosson</namePart><affiliation>UMR 7009 CNRS – Univ. Paris VI, Observatore de Villefranche/mer, Marine Station, Villefranche‐sur‐mer, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">S. M. H.</namePart><namePart type="family">Alavi</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">M.</namePart><namePart type="family">Rodina</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">V.</namePart><namePart type="family">Kaspar</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">D.</namePart><namePart type="family">Gela</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">O.</namePart><namePart type="family">Linhart</namePart><affiliation>Research Institute of Fish Culture and Hydrobiology, University of South Bohemia, Vodnany, Czech Republic</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">A.</namePart><namePart type="family">Ciereszko</namePart><affiliation>Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article"></genre><originInfo><publisher>Blackwell Publishing Ltd</publisher><place><placeTerm type="text">Oxford, UK</placeTerm></place><dateIssued encoding="w3cdtf">2008-08</dateIssued><edition>Received: September 15, 2007 Accepted: March 12, 2008</edition><copyrightDate encoding="w3cdtf">2008</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType><extent unit="figures">3</extent></physicalDescription><abstract lang="en">Trypsin‐like activity, similar to acrosin, is present in sturgeon spermatozoa and can be a potential target for trypsin inhibitors. The objective of this work was to use a fluorescent soybean trypsin inhibitor (SBTI) conjugate with the Alexa Fluor® 488 dye for visualization of the sturgeon acrosome. After incubation with SBTI‐Alexa, a strong signal was observed both in the acrosome and midpiece or implantation fossa region. We have also found that SBTI‐Alexa staining can be combined with PI viability test. Detailed examination of staining pattern revealed that SBTI‐Alexa can stain either acrosome or whole sperm. Staining of whole sperm correlated with dead staining (r2 = 0.94, P < 0.01). However, in fresh semen most cells (93–97%) were not stained with SBTI‐Alexa, probably due to intact acrosomes. Further studies should test if SBTI‐Alexa can be applied to monitor the acrosome status during the acrosome reaction and cryopreservation.</abstract><relatedItem type="host"><titleInfo><title>Journal of Applied Ichthyology</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0175-8659</identifier><identifier type="eISSN">1439-0426</identifier><identifier type="DOI">10.1111/(ISSN)1439-0426</identifier><identifier type="PublisherID">JAI</identifier><part><date>2008</date><detail type="volume"><caption>vol.</caption><number>24</number></detail><detail type="issue"><caption>no.</caption><number>4</number></detail><extent unit="pages"><start>514</start><end>516</end><total>3</total></extent></part></relatedItem><identifier type="istex">D410D188F9BE140E2E47C8262B3AE00BE1D95A10</identifier><identifier type="DOI">10.1111/j.1439-0426.2008.01140.x</identifier><identifier type="ArticleID">JAI1140</identifier><accessCondition type="use and reproduction" contentType="copyright">© 2008 Blackwell Verlag, Berlin</accessCondition><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>Blackwell Publishing Ltd</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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