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Reproductive Biology Section, Departments of Obstetrics/Gynecology and Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520
Address all correspondence and requests for reprints to: Dr. Pinar H. Kodaman, Reproductive Biology Section, Department of Obstetrics and Gynecology, Yale University School of Medicine, P.O. Box 208063, New Haven, Connecticut 06520-8063. E-mail: kodamaph{at}biomed.med.yale.edu
The ovulatory LH surge results in follicular inflammation with an
increase in cytokines and PGs. Reactive oxygen species (ROS) are also
produced during inflammatory processes. To study ROS generation during
the ovulatory cascade, preovulatory follicles were dissected from
immature female rats primed with PMSG. Follicles were isolated, and ROS
generation was assessed by luminol-amplified chemiluminescence.
Immature rat granulosa cells were also subjected to luminometry after
isolation from immature rats treated with diethylstilbestrol. Phorbol
ester-stimulated ROS generation by follicular cells was completely
suppressed by superoxide dismutase and the NADPH/NADH oxidase inhibitor
diphenylene iodonium bisulfate, whereas catalase was without effect.
Fractionation of granulosa cells with an antibody against leukocyte
common antigen-1 showed that leukocyte-enriched cells produced more
than 95% of the superoxide measured. In vivo treatment
with LH produced a 5-fold increase in phorbol-stimulated superoxide
production by isolated follicles. This response was maximal within
4 h and was blocked by indomethacin. In vivo
administration of PGE2 and PGF2
did not
reverse the blockade by indomethacin; however, isolated follicles
incubated with PGE2 produced a time-dependent increase in
phorbol-stimulated superoxide generation. Thus, a superoxide generator
is present in the preovulatory follicle that is leukocytic in origin,
hormone regulated, and activated by a protein kinase C-dependent
pathway. The regulated generation of superoxide by preovulatory
follicles may indicate a role for ROS in the periovulatory period.
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