Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa
Contents The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be...
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| Veröffentlicht in: | Reproduction in domestic animals 2015-08, Vol.50 (4), p.688-691 |
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| creator | Casao, A Mata-Campuzano, M Ordás, L Cebrián-Pérez, JA Muiño-Blanco, T Martínez-Pastor, F |
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The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p |
| doi_str_mv | 10.1111/rda.12549 |
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The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p < 0.001), and a 12‐h incubation increased cPARP similarly in both fractions (p < 0.001). cPARP seems to be an early marker of apoptosis in ram semen, although its presence in untreated samples was weakly related to worse quality (pellet/interface). We suggest to study the relationship of PARP and cPARP levels with between‐male differences on sperm fertility.</description><identifier>ISSN: 0936-6768</identifier><identifier>EISSN: 1439-0531</identifier><identifier>DOI: 10.1111/rda.12549</identifier><identifier>PMID: 26031316</identifier><language>eng</language><publisher>Germany: Blackwell Publishing Ltd</publisher><subject>Acrosome - enzymology ; Animal reproduction ; Animals ; Apoptosis ; Biomarkers ; Caspase 3 - metabolism ; Caspases - metabolism ; Enzyme Activation ; Flow Cytometry - veterinary ; Fluorescent Antibody Technique, Indirect - veterinary ; Male ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases - analysis ; Poly(ADP-ribose) Polymerases - metabolism ; Sheep ; Sperm ; Spermatogenesis ; Spermatozoa - enzymology ; Spermatozoa - physiology</subject><ispartof>Reproduction in domestic animals, 2015-08, Vol.50 (4), p.688-691</ispartof><rights>2015 Blackwell Verlag GmbH</rights><rights>2015 Blackwell Verlag GmbH.</rights><rights>Copyright © 2015 Blackwell Verlag GmbH</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4969-7cb09a81cd26ee858670f8ca5ad0ad0a7d2b2c5bf9bb74c61408c2e47fbdfacc3</citedby><cites>FETCH-LOGICAL-c4969-7cb09a81cd26ee858670f8ca5ad0ad0a7d2b2c5bf9bb74c61408c2e47fbdfacc3</cites><orcidid>0000-0003-2987-4302</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Frda.12549$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Frda.12549$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26031316$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Casao, A</creatorcontrib><creatorcontrib>Mata-Campuzano, M</creatorcontrib><creatorcontrib>Ordás, L</creatorcontrib><creatorcontrib>Cebrián-Pérez, JA</creatorcontrib><creatorcontrib>Muiño-Blanco, T</creatorcontrib><creatorcontrib>Martínez-Pastor, F</creatorcontrib><title>Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa</title><title>Reproduction in domestic animals</title><addtitle>Reprod Dom Anim</addtitle><description>Contents
The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p < 0.001), and a 12‐h incubation increased cPARP similarly in both fractions (p < 0.001). cPARP seems to be an early marker of apoptosis in ram semen, although its presence in untreated samples was weakly related to worse quality (pellet/interface). We suggest to study the relationship of PARP and cPARP levels with between‐male differences on sperm fertility.</description><subject>Acrosome - enzymology</subject><subject>Animal reproduction</subject><subject>Animals</subject><subject>Apoptosis</subject><subject>Biomarkers</subject><subject>Caspase 3 - metabolism</subject><subject>Caspases - metabolism</subject><subject>Enzyme Activation</subject><subject>Flow Cytometry - veterinary</subject><subject>Fluorescent Antibody Technique, Indirect - veterinary</subject><subject>Male</subject><subject>Poly (ADP-Ribose) Polymerase-1</subject><subject>Poly(ADP-ribose) Polymerases - analysis</subject><subject>Poly(ADP-ribose) Polymerases - metabolism</subject><subject>Sheep</subject><subject>Sperm</subject><subject>Spermatogenesis</subject><subject>Spermatozoa - enzymology</subject><subject>Spermatozoa - physiology</subject><issn>0936-6768</issn><issn>1439-0531</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10FtLwzAYBuAgis7DhX9ACt4orFuSNklzOTadwtQxp16GNP0K1XatSefp15s59UIwBALh-V6SF6FDgnvEr77NdI9QFssN1CFxJEPMIrKJOlhGPOSCJzto17lHjAlLhNhGO5TjiESEd9B4WIJ-gSyYDmbTkHQDvQgGTd20dVuY4ErbJ7DdwPjbFIIRtGBaj4tFMNNVcNuArXRbf9R6H23lunRw8H3uobvzs_nwIpzcjC-Hg0loYsllKEyKpU6IySgHSFjCBc4To5nO8GqLjKbUsDSXaSpiw0mME0MhFnma5dqYaA-drHMbWz8vwbWqKpyBstQLqJdOES4FxZJx4enxH_pYL-3Cv26lOOYUx7FXp2tlbO2chVw1tqi0fVcEq1W7yrervtr19ug7cZlWkP3Knzo96K_Ba1HC-_9JajYa_ESG64nCtfD2O-F7V_4LgqmH67Ga0rkULBLqPvoE01WQOQ</recordid><startdate>201508</startdate><enddate>201508</enddate><creator>Casao, A</creator><creator>Mata-Campuzano, M</creator><creator>Ordás, L</creator><creator>Cebrián-Pérez, JA</creator><creator>Muiño-Blanco, T</creator><creator>Martínez-Pastor, F</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-2987-4302</orcidid></search><sort><creationdate>201508</creationdate><title>Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa</title><author>Casao, A ; Mata-Campuzano, M ; Ordás, L ; Cebrián-Pérez, JA ; Muiño-Blanco, T ; Martínez-Pastor, F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4969-7cb09a81cd26ee858670f8ca5ad0ad0a7d2b2c5bf9bb74c61408c2e47fbdfacc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Acrosome - enzymology</topic><topic>Animal reproduction</topic><topic>Animals</topic><topic>Apoptosis</topic><topic>Biomarkers</topic><topic>Caspase 3 - metabolism</topic><topic>Caspases - metabolism</topic><topic>Enzyme Activation</topic><topic>Flow Cytometry - veterinary</topic><topic>Fluorescent Antibody Technique, Indirect - veterinary</topic><topic>Male</topic><topic>Poly (ADP-Ribose) Polymerase-1</topic><topic>Poly(ADP-ribose) Polymerases - analysis</topic><topic>Poly(ADP-ribose) Polymerases - metabolism</topic><topic>Sheep</topic><topic>Sperm</topic><topic>Spermatogenesis</topic><topic>Spermatozoa - enzymology</topic><topic>Spermatozoa - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Casao, A</creatorcontrib><creatorcontrib>Mata-Campuzano, M</creatorcontrib><creatorcontrib>Ordás, L</creatorcontrib><creatorcontrib>Cebrián-Pérez, JA</creatorcontrib><creatorcontrib>Muiño-Blanco, T</creatorcontrib><creatorcontrib>Martínez-Pastor, F</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Reproduction in domestic animals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Casao, A</au><au>Mata-Campuzano, M</au><au>Ordás, L</au><au>Cebrián-Pérez, JA</au><au>Muiño-Blanco, T</au><au>Martínez-Pastor, F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa</atitle><jtitle>Reproduction in domestic animals</jtitle><addtitle>Reprod Dom Anim</addtitle><date>2015-08</date><risdate>2015</risdate><volume>50</volume><issue>4</issue><spage>688</spage><epage>691</epage><pages>688-691</pages><issn>0936-6768</issn><eissn>1439-0531</eissn><abstract>Contents
The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p < 0.001), and a 12‐h incubation increased cPARP similarly in both fractions (p < 0.001). cPARP seems to be an early marker of apoptosis in ram semen, although its presence in untreated samples was weakly related to worse quality (pellet/interface). We suggest to study the relationship of PARP and cPARP levels with between‐male differences on sperm fertility.</abstract><cop>Germany</cop><pub>Blackwell Publishing Ltd</pub><pmid>26031316</pmid><doi>10.1111/rda.12549</doi><tpages>4</tpages><orcidid>https://orcid.org/0000-0003-2987-4302</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Acrosome - enzymology Animal reproduction Animals Apoptosis Biomarkers Caspase 3 - metabolism Caspases - metabolism Enzyme Activation Flow Cytometry - veterinary Fluorescent Antibody Technique, Indirect - veterinary Male Poly (ADP-Ribose) Polymerase-1 Poly(ADP-ribose) Polymerases - analysis Poly(ADP-ribose) Polymerases - metabolism Sheep Sperm Spermatogenesis Spermatozoa - enzymology Spermatozoa - physiology |
| title | Cleaved PARP-1, an Apoptotic Marker, can be Detected in Ram Spermatozoa |
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