caa a22 4500 445318465 CHVBK 20180317142655.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s10126-010-9324-9 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9324-9 Ca2+, Mg2+-dependent DNase Involvement in Apoptotic Effects in Spermatozoa of Sea Urchin Strongylocentrotus intermedius Induced by Two-Headed Sphingolipid Rhizochalin [Elektronische Daten] [Juriy Sibirtsev, Valeria Shastina, Natalia Menzorova, Tatyana Makarieva, Valeriy Rasskazov] Previously, we have purified three distinct DNases from spermatozoa of sea urchin Strongylocentrotus intermedius and we suppose the role of Ca2+, Mg2+-dependent DNase (Ca, Mg-DNase) in apoptosis of spermatozoa. Two-headed sphingolipid rhizochalin (Rhz) induced characteristic apoptotic nuclear chromatin changes, internucleosomal DNA cleavage, and activation of caspase-9, caspase-8, and caspase-3 in spermatozoa as was shown by fluorescence Hoechst 33342/PI/FDA analysis, DNA fragmentation assay, and fluorescence caspase inhibitors FAM-LEHD-fmk, FAM-IETD-fmk, and FAM-DEVD-fmk, respectively. Inhibitor of caspase-3 z-DEVD-fmk subdued Rhz-induced internucleosomal ladder formation, which confirmed the major role of caspase-3 in apoptotic DNA cleavage probably through Ca, Mg-DNase activation. Participation of sea urchin Ca, Mg-DNase in apoptosis of spermatozoa was demonstrated by ions Zn2+ blocking of Rhz-induced DNA fragmentation due to direct inhibition of the Ca, Mg-DNase and internucleosomal cleavage of HeLa S and Vero E6 cell nuclei chromatin by highly purified Ca, Mg-DNase. Springer Science+Business Media, LLC, 2010 Sea urchin nationallicence Spermatozoa nationallicence Ca2+ nationallicence Mg2+-dependent DNase nationallicence Two-headed sphingolipids nationallicence Apoptosis nationallicence Caspase nationallicence EGTA : Ethyleneglycoltetraacetic acid nationallicence EDTA : Ethylenediaminetetraacetic acid nationallicence DTT : Dethiothreithol nationallicence PMSF : Phenylmethylsulfonyl fluoride nationallicence SDS : Sodium dodecyl sulfate nationallicence PI : Propidium iodide nationallicence FDA : Fluorescein diacetate nationallicence FAM-LEHD-fmk : Carboxyfluorescein analog of benzyloxycarbonyl-Leu-Glu-His-Asp(OMe) fluoromethyl ketone (z-LEHD-fmk) nationallicence FAM-IETD-fmk : Carboxyfluorescein analog of benzyloxycarbonyl-Ile-Glu-Thr-Asp(OMe)fluoromethyl ketone (z-IETD-fmk) nationallicence FAM-DEVD-fmk : Carboxyfluorescein analog of benzyloxycarbonyl-Asp(OMe)-Glu(O-Me)-Val-Asp(O-Me) fluoromethyl ketone (z-DEVD-fmk) nationallicence Sibirtsev Juriy Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut Shastina Valeria Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut Menzorova Natalia Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut Makarieva Tatyana Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut Rasskazov Valeriy Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut Marine Biotechnology Springer-Verlag 13/3(2011-06-01), 536-543 1436-2228 13:3<536 2011 13 10126 https://doi.org/10.1007/s10126-010-9324-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9324-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sibirtsev Juriy Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut NATIONALLICENCE 700 1- Shastina Valeria Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut NATIONALLICENCE 700 1- Menzorova Natalia Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut NATIONALLICENCE 700 1- Makarieva Tatyana Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut NATIONALLICENCE 700 1- Rasskazov Valeriy Laboratory of Marine Biochemistry, Pacific Institute of Bioorganic Chemistry, Far East Division of Russian Academy of Sciences, 690022, pr. 100 let Vladivostoku, 159, Vladivostok, Russian Federation aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/3(2011-06-01), 536-543 1436-2228 13:3<536 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer