Molecular requirements for doxorubicin-mediated death in murine oocytes.
We previously published evidence that oocytes exposed to doxorubicin (DXR), a widely used chemotherapeutic agent, rapidly undergo morphological and biochemical changes via discrete effector signaling pathways consistent with the occurrence of apoptosis. In this report, we elucidated the molecular requirements for actions of this drug in oocytes. Our results indicate that within 1 h of exposure DXR causes rapid DNA damage, and commits the oocyte to cytoplasmic fragmentation by the fourth hour, followed by delayed oocyte activation and execution of cytoplasmic fragmentation. Inhibitors that interfere with oocyte activation consistently rescue cytoplasmic fragmentation, but fail to suppress DNA damage. There was evidence of depletion of Bax, Caspase-2, MA-3 and Bcl-x transcripts, suggesting that modulations by DXR caused recruitment of these maternal transcripts into the translation process. Furthermore, sphingolipids such as sphingosine-1-phosphate and ceramide modulate DXR actions by, respectively, altering its intracellular trafficking, or by sustaining the drug's contact with DNA.
Jurisicova
A
A
Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114, USA.
Lee
H-J
HJ
D'Estaing
S G
SG
Tilly
J
J
Perez
G I
GI
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000378:metabolism / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D053143:Caspase 2 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D002478:Cells, Cultured; D004249:DNA Damage / Q000187:drug effects*; D004317:Doxorubicin / Q000378:metabolism / Q000494:pharmacology*; D005260:Female; D008246:Lysophospholipids / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D014176:Protein Biosynthesis / Q000187:drug effects; D013110:Sphingosine / Q000031:analogs & derivatives / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051020:bcl-X Protein / Q000378:metabolism
D000903:Antibiotics, Antineoplastic; C494827:Bax protein, mouse; C494812:Bcl2l1 protein, mouse; D008246:Lysophospholipids; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; C060506:sphingosine 1-phosphate; D004317:Doxorubicin; D053143:Caspase 2; D013110:Sphingosine