In Situ Staining for Poly(ADPRibose) Polymerase Activity Using an NAD Analogue
Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is...
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| Veröffentlicht in: | The journal of histochemistry and cytochemistry 1998-11, Vol.46 (11), p.1279-1289 |
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| container_title | The journal of histochemistry and cytochemistry |
| container_volume | 46 |
| creator | Davis, R. Eric Mysore, Veena Browning, Jared C Hsieh, Joseph C Lu, Quynh-Anh T Katsikis, Peter D |
| description | Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is not fully understood. We have modified an existing in situ method for detection of PARP activity by using an NAD analogue in which adenine is modified by an “etheno” (vinyl) bridge. Etheno-NAD serves as a PARP substrate in an initial enzymatic reaction; a specific antibody to ethenoadenosine is then used in an immunohistochemical reaction to detect the production of modified poly(ADP-ribose). The method produces strong and specific labeling of nuclei in which PARP has been activated, i.e., those in which DNA strand breaks have been produced, and the results can be analyzed by microscopy, flow cytometry, or colorimetry. The method is applicable to cultured cells in several formats and to frozen tissue sections. The particular characteristics of the new method may assist in future in situ studies of PARP activation. |
| doi_str_mv | 10.1177/002215549804601108 |
| format | Article |
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Eric ; Mysore, Veena ; Browning, Jared C ; Hsieh, Joseph C ; Lu, Quynh-Anh T ; Katsikis, Peter D</creator><creatorcontrib>Davis, R. Eric ; Mysore, Veena ; Browning, Jared C ; Hsieh, Joseph C ; Lu, Quynh-Anh T ; Katsikis, Peter D</creatorcontrib><description>Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is not fully understood. We have modified an existing in situ method for detection of PARP activity by using an NAD analogue in which adenine is modified by an “etheno” (vinyl) bridge. Etheno-NAD serves as a PARP substrate in an initial enzymatic reaction; a specific antibody to ethenoadenosine is then used in an immunohistochemical reaction to detect the production of modified poly(ADP-ribose). The method produces strong and specific labeling of nuclei in which PARP has been activated, i.e., those in which DNA strand breaks have been produced, and the results can be analyzed by microscopy, flow cytometry, or colorimetry. The method is applicable to cultured cells in several formats and to frozen tissue sections. The particular characteristics of the new method may assist in future in situ studies of PARP activation.</description><identifier>ISSN: 0022-1554</identifier><identifier>EISSN: 1551-5044</identifier><identifier>DOI: 10.1177/002215549804601108</identifier><identifier>PMID: 9774627</identifier><language>eng</language><publisher>Los Angeles, CA: Histochemical Soc</publisher><subject>Animals ; Benzamides - pharmacology ; Brain - drug effects ; Brain - enzymology ; Brain - radiation effects ; Bromodeoxyuridine - pharmacology ; Cell Line ; Humans ; Immunoenzyme Techniques ; Methylnitronitrosoguanidine - pharmacology ; Mice ; NAD - analogs & derivatives ; NAD - pharmacology ; Nitroprusside - pharmacology ; Poly(ADP-ribose) Polymerases - metabolism ; Pyrogallol - pharmacology ; Rats ; Staining and Labeling - methods ; Ultraviolet Rays</subject><ispartof>The journal of histochemistry and cytochemistry, 1998-11, Vol.46 (11), p.1279-1289</ispartof><rights>1998 Authors</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c414t-53bb6a04ad5ed6a7a0ecf0c2717f68d39649745381cb8b3380967f7a5a7469363</citedby><cites>FETCH-LOGICAL-c414t-53bb6a04ad5ed6a7a0ecf0c2717f68d39649745381cb8b3380967f7a5a7469363</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1177/002215549804601108$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1177/002215549804601108$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>314,776,780,21798,27901,27902,43597,43598</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9774627$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Davis, R. Eric</creatorcontrib><creatorcontrib>Mysore, Veena</creatorcontrib><creatorcontrib>Browning, Jared C</creatorcontrib><creatorcontrib>Hsieh, Joseph C</creatorcontrib><creatorcontrib>Lu, Quynh-Anh T</creatorcontrib><creatorcontrib>Katsikis, Peter D</creatorcontrib><title>In Situ Staining for Poly(ADPRibose) Polymerase Activity Using an NAD Analogue</title><title>The journal of histochemistry and cytochemistry</title><addtitle>J Histochem Cytochem</addtitle><description>Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is not fully understood. We have modified an existing in situ method for detection of PARP activity by using an NAD analogue in which adenine is modified by an “etheno” (vinyl) bridge. Etheno-NAD serves as a PARP substrate in an initial enzymatic reaction; a specific antibody to ethenoadenosine is then used in an immunohistochemical reaction to detect the production of modified poly(ADP-ribose). The method produces strong and specific labeling of nuclei in which PARP has been activated, i.e., those in which DNA strand breaks have been produced, and the results can be analyzed by microscopy, flow cytometry, or colorimetry. The method is applicable to cultured cells in several formats and to frozen tissue sections. The particular characteristics of the new method may assist in future in situ studies of PARP activation.</description><subject>Animals</subject><subject>Benzamides - pharmacology</subject><subject>Brain - drug effects</subject><subject>Brain - enzymology</subject><subject>Brain - radiation effects</subject><subject>Bromodeoxyuridine - pharmacology</subject><subject>Cell Line</subject><subject>Humans</subject><subject>Immunoenzyme Techniques</subject><subject>Methylnitronitrosoguanidine - pharmacology</subject><subject>Mice</subject><subject>NAD - analogs & derivatives</subject><subject>NAD - pharmacology</subject><subject>Nitroprusside - pharmacology</subject><subject>Poly(ADP-ribose) Polymerases - metabolism</subject><subject>Pyrogallol - pharmacology</subject><subject>Rats</subject><subject>Staining and Labeling - methods</subject><subject>Ultraviolet Rays</subject><issn>0022-1554</issn><issn>1551-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtKw0AUhgdRaq2-gCBk5WURO5NM5rIMrZdC0WLtepgkk3ZKLjqTGPoiPpBPZmKKG8HVgf98_885PwDnCN4iROkYQs9DQYA5g5hAhCA7AMNWQG4AMT4Eww5wO-IYnFi7hRBhHLABGHBKMfHoECxmhbPUVe0sK6kLXaydtDTOosx21-F08fX5oqPSqpsfJVdGWuWEcaU_dLVzVrbjZeE8hVMnLGRWrmt1Co5SmVl1tp8jsLq_e508uvPnh9kknLsxRrhyAz-KiIRYJoFKiKQSqjiFsUcRTQlLfE4wpzjwGYojFvk-g5zQlMpAtodzn_gjcNnnvpnyvVa2Erm2scoyWaiytoJwThjjuAW9HoxNaa1RqXgzOpdmJxAUXY3ib42t6WKfXke5Sn4t-97a_bjfW7lWYlvWpn3f_p941Ts2er1ptFHC5jLL2nwkmqbBpHUK5FHufwMLwoYo</recordid><startdate>19981101</startdate><enddate>19981101</enddate><creator>Davis, R. Eric</creator><creator>Mysore, Veena</creator><creator>Browning, Jared C</creator><creator>Hsieh, Joseph C</creator><creator>Lu, Quynh-Anh T</creator><creator>Katsikis, Peter D</creator><general>Histochemical Soc</general><general>SAGE Publications</general><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>7X8</scope></search><sort><creationdate>19981101</creationdate><title>In Situ Staining for Poly(ADPRibose) Polymerase Activity Using an NAD Analogue</title><author>Davis, R. Eric ; Mysore, Veena ; Browning, Jared C ; Hsieh, Joseph C ; Lu, Quynh-Anh T ; Katsikis, Peter D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c414t-53bb6a04ad5ed6a7a0ecf0c2717f68d39649745381cb8b3380967f7a5a7469363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Animals</topic><topic>Benzamides - pharmacology</topic><topic>Brain - drug effects</topic><topic>Brain - enzymology</topic><topic>Brain - radiation effects</topic><topic>Bromodeoxyuridine - pharmacology</topic><topic>Cell Line</topic><topic>Humans</topic><topic>Immunoenzyme Techniques</topic><topic>Methylnitronitrosoguanidine - pharmacology</topic><topic>Mice</topic><topic>NAD - analogs & derivatives</topic><topic>NAD - pharmacology</topic><topic>Nitroprusside - pharmacology</topic><topic>Poly(ADP-ribose) Polymerases - metabolism</topic><topic>Pyrogallol - pharmacology</topic><topic>Rats</topic><topic>Staining and Labeling - methods</topic><topic>Ultraviolet Rays</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Davis, R. Eric</creatorcontrib><creatorcontrib>Mysore, Veena</creatorcontrib><creatorcontrib>Browning, Jared C</creatorcontrib><creatorcontrib>Hsieh, Joseph C</creatorcontrib><creatorcontrib>Lu, Quynh-Anh T</creatorcontrib><creatorcontrib>Katsikis, Peter D</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The journal of histochemistry and cytochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Davis, R. Eric</au><au>Mysore, Veena</au><au>Browning, Jared C</au><au>Hsieh, Joseph C</au><au>Lu, Quynh-Anh T</au><au>Katsikis, Peter D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In Situ Staining for Poly(ADPRibose) Polymerase Activity Using an NAD Analogue</atitle><jtitle>The journal of histochemistry and cytochemistry</jtitle><addtitle>J Histochem Cytochem</addtitle><date>1998-11-01</date><risdate>1998</risdate><volume>46</volume><issue>11</issue><spage>1279</spage><epage>1289</epage><pages>1279-1289</pages><issn>0022-1554</issn><eissn>1551-5044</eissn><abstract>Poly(ADP-ribose) polymerase (PARP) is a highly abundant nuclear enzyme which metabolizes NAD, in response to DNA strand breakage, to produce chains of poly(ADP-ribose) attached to nuclear proteins. PARP activation has been implicated in ischemia/reperfusion injury, but its biological significance is not fully understood. 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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; SAGE Complete; Free Full-Text Journals in Chemistry |
| subjects | Animals Benzamides - pharmacology Brain - drug effects Brain - enzymology Brain - radiation effects Bromodeoxyuridine - pharmacology Cell Line Humans Immunoenzyme Techniques Methylnitronitrosoguanidine - pharmacology Mice NAD - analogs & derivatives NAD - pharmacology Nitroprusside - pharmacology Poly(ADP-ribose) Polymerases - metabolism Pyrogallol - pharmacology Rats Staining and Labeling - methods Ultraviolet Rays |
| title | In Situ Staining for Poly(ADPRibose) Polymerase Activity Using an NAD Analogue |
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