Ca2+, Mg2+-dependent DNase Involvement in Apoptotic Effects in Spermatozoa of Sea Urchin Strongylocentrotus intermedius Induced by Two-Headed Sphingolipid Rhizochalin

Previously, we have purified three distinct DNases from spermatozoa of sea urchin Strongylocentrotus intermedius and we suppose the role of Ca 2+ , Mg 2+ -dependent DNase (Ca, Mg-DNase) in apoptosis of spermatozoa. Two-headed sphingolipid rhizochalin (Rhz) induced characteristic apoptotic nuclear ch...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2011-06, Vol.13 (3), p.536-543
Main Authors: Sibirtsev, Juriy T., Shastina, Valeria V., Menzorova, Natalia I., Makarieva, Tatyana N., Rasskazov, Valeriy A.
Format: Article
Language:English
Subjects:
Acids
Animals
Apoptosis
Apoptosis - physiology
Biomedical and Life Sciences
Biotechnology
Calcium - metabolism
Caspase 3 - metabolism
Deoxyribonucleases - metabolism
Deoxyribonucleic acid
DNA
DNA Fragmentation
Engineering
Fluorescence
Fluorides
Freshwater & Marine Ecology
Glycosphingolipids - metabolism
Invertebrates
Life Sciences
Magnesium - metabolism
Male
Marine biology
Microbiology
Original Article
Reproductive system
Seawater
Sperm
Spermatozoa - metabolism
Strongylocentrotus - enzymology
Studies
Zoology
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Summary:Previously, we have purified three distinct DNases from spermatozoa of sea urchin Strongylocentrotus intermedius and we suppose the role of Ca 2+ , Mg 2+ -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 Zn 2+ 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.
ISSN:1436-2228
1436-2236
DOI:10.1007/s10126-010-9324-9