We report a short-term culture system that allows to define novel characteristic of programmed cell death (PCD) in fetal oocytes and to underscore new aspects of this process. Mouse fetal oocytes cultured in conditions allowing meiotic prophase I progression underwent apoptotic degeneration waves as revealed by TUNEL staining. TEM observations revealed recurrent atypical apoptotic morphologies characterized by the absence of chromatin margination and nuclear fragmentation; oocytes with autophagic and necrotic features were also observed. Further characterization of oocyte death evidenced DNA ladder, Annexin V binding, PARP cleavage, and usually caspase activation (namely caspase-2). In the aim to modulate the oocyte death process, we found that the addition to the culture medium of the pan-caspase inhibitors Z-VAD or caspase-2-specific inhibitor Z-VDVAD resulted in a partial and transient prevention of this process. Oocyte death was significantly reduced by the antioxidant agent NAC and partly prevented by KL and IGF-I growth factors. Finally, oocyte apoptosis was reduced by calpain inhibitor I and increased by rapamycin after prolonged culture. These results support the notion that fetal oocytes undergo degeneration mostly by apoptosis. This process is, however, often morphologically atypical and encompasses other forms of cell death including caspase-independent apoptosis and autophagia. The observation that oocyte death occurs mainly at certain stages of meiosis and can only be attenuated by typical anti-apoptotic treatments favors the notion that it is controlled at least in part by stage-specific oocyte-autonomous meiotic checkpoints and when activated is little amenable to inhibition being the oocyte able to switch back and forth among different death pathways.

Lobascio, A., Klinger, F.g., Scaldaferri, M., Farini, D., DE FELICI, M. (2007). Analysis of programmed cell death in mouse fetal oocytes. REPRODUCTION, 134(2), 241-252 [10.1530/REP-07-0141].

Analysis of programmed cell death in mouse fetal oocytes

KLINGER, FRANCESCA GIOIA;FARINI, DONATELLA;DE FELICI, MASSIMO
2007

Abstract

We report a short-term culture system that allows to define novel characteristic of programmed cell death (PCD) in fetal oocytes and to underscore new aspects of this process. Mouse fetal oocytes cultured in conditions allowing meiotic prophase I progression underwent apoptotic degeneration waves as revealed by TUNEL staining. TEM observations revealed recurrent atypical apoptotic morphologies characterized by the absence of chromatin margination and nuclear fragmentation; oocytes with autophagic and necrotic features were also observed. Further characterization of oocyte death evidenced DNA ladder, Annexin V binding, PARP cleavage, and usually caspase activation (namely caspase-2). In the aim to modulate the oocyte death process, we found that the addition to the culture medium of the pan-caspase inhibitors Z-VAD or caspase-2-specific inhibitor Z-VDVAD resulted in a partial and transient prevention of this process. Oocyte death was significantly reduced by the antioxidant agent NAC and partly prevented by KL and IGF-I growth factors. Finally, oocyte apoptosis was reduced by calpain inhibitor I and increased by rapamycin after prolonged culture. These results support the notion that fetal oocytes undergo degeneration mostly by apoptosis. This process is, however, often morphologically atypical and encompasses other forms of cell death including caspase-independent apoptosis and autophagia. The observation that oocyte death occurs mainly at certain stages of meiosis and can only be attenuated by typical anti-apoptotic treatments favors the notion that it is controlled at least in part by stage-specific oocyte-autonomous meiotic checkpoints and when activated is little amenable to inhibition being the oocyte able to switch back and forth among different death pathways.
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/17
English
Con Impact Factor ISI
Sirolimus; Fetus; Animals; Apoptosis; Antioxidants; Autophagy; Ovary; Stem Cell Factor; Microscopy, Electron, Transmission; Necrosis; Acetylcysteine; Oocytes; Glycoproteins; DNA Fragmentation; Oligopeptides; Meiotic Prophase I; Cell Culture Techniques; Caspase 2; Mice; Insulin-Like Growth Factor I; Immunosuppressive Agents; In Situ Nick-End Labeling; Mice, Inbred Strains; Cells, Cultured; Cysteine Proteinase Inhibitors; Female
Lobascio, A., Klinger, F.g., Scaldaferri, M., Farini, D., DE FELICI, M. (2007). Analysis of programmed cell death in mouse fetal oocytes. REPRODUCTION, 134(2), 241-252 [10.1530/REP-07-0141].
Lobascio, A; Klinger, Fg; Scaldaferri, M; Farini, D; DE FELICI, M
Articolo su rivista
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/44613
Citazioni
  • ???jsp.display-item.citation.pmc??? 24
  • Scopus ND
  • ???jsp.display-item.citation.isi??? 58
social impact