Expression and localization of Fas ligand and Fas during atresia in porcine ovarian follicles.
To reveal the mechanisms regulating the selective atresia of follicles in porcine ovaries, we examined the changes in the mRNA and protein levels of cell-death ligand, Fas/APO-1/CD95 ligand (FasL), and its receptor, Fas/APO-1/CD95 (Fas), and the localization of the proteins in granulosa cells during follicular atresia using the reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical techniques, respectively. Trace levels of FasL mRNA and protein were detected in the granulosa cells of healthy follicles; however, weak levels were detected in those of early atretic follicles, and the levels increased during atresia. Trace/weak levels of Fas mRNA and protein were detected in the granulosa cells of healthy follicles. Fas protein was located in the cytoplasmic area, not in cell membrane area, indicating that it has no activity in regard to inducing apoptosis. When apoptosis commences in granulosa cells, Fas moves from the cytoplasmic to cell membrane area. FasL and Fas mRNAs and proteins in granulosa cells were upregulated during follicular atresia. The FasL and Fas system may play a crucial role in the regulation of apoptosis in granulosa cells during selective follicular atresia in porcine ovaries.
Inoue
Naoko
N
Laboratory of Animal Morphology and Function, Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Maeda
Akihisa
A
Matsuda-Minehata
Fuko
F
Fukuta
Katsuhiro
K
Manabe
Noboru
N
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D053222:Fas Ligand Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005496:Follicular Atresia / Q000235:genetics / Q000378:metabolism*; D006107:Granulosa Cells / Q000378:metabolism; D007150:Immunohistochemistry / Q000662:veterinary; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA; D013552:Swine / Q000235:genetics / Q000378:metabolism*; D019014:fas Receptor / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D053222:Fas Ligand Protein; D012333:RNA, Messenger; D019014:fas Receptor