title: Involvement of the sphingolipid ceramide in heat-shock-induced apoptosis of bovine oocytes.
abstract: Programmed cell death via the sphingomyelin pathway has been suggested to underlie heat-shock disturbance of oocyte developmental competence. A series of experiments were performed to characterise the role of the sphingolipid ceramide in heat-shock-induced apoptosis, and to determine whether ceramide formation can be regulated. Bovine cumulus-oocyte complexes (COCs) were aspirated from ovaries collected in the cold season (November-April), in vitro-matured, fertilised and cultured for 8 days. Exposure of COCs to heat shock (41°C) during maturation reduced cleavage rate and blastocyst formation relative to the control group (38.5°C). Annexin-V binding (V-FITC assay), which is associated with the early apoptotic event of membrane phosphatidylserine turnover, was higher in oocytes exposed to short-term versus long-term heat shock, suggesting that heat-shock-induced apoptosis involves membrane alterations. Similar to heat exposure, oocyte maturation with C(2)-ceramide had a dose-dependent deleterious effect on the first cleavages and subsequent embryonic development in association with increased annexin-V binding. Blocking endogenous ceramide generation with fumonisin B1, a specific inhibitor of dihydroceramide synthase (i.e. de novo formation), moderated, to some extent, the effects of heat shock on oocyte developmental competence, suggesting that ceramide plays an important role in heat-shock-induced apoptosis.
authors.affiliation: Department of Animal Sciences, The Hebrew University, Rehovot, Israel.
authors.forename: Dorit
authors.initials: D
authors.lastname: Kalo
authors.forename: Zvi
authors.initials: Z
authors.lastname: Roth
mesh_terms: D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D002518:Ceramides / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054885:Cumulus Cells / Q000502:physiology; D053938:DNA Fragmentation; D004477:Ectogenesis / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D037341:Fumonisins / Q000494:pharmacology; D018869:Heat-Shock Response; D006358:Hot Temperature / Q000009:adverse effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D009866:Oogenesis / Q000187:drug effects; D009994:Osmolar Concentration; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D010718:Phosphatidylserines / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D013997:Time Factors
chemical_list: D002518:Ceramides; D004791:Enzyme Inhibitors; D037341:Fumonisins; C064769:N-acetylsphingosine; D010718:Phosphatidylserines; D013107:Sphingolipids; C056933:fumonisin B1; D010088:Oxidoreductases; C473786:dihydroceramide synthase; D013110:Sphingosine