Occurrence of immunoreactive Met- and Leu-enkephalin-like peptides in the ovary of the green frog, Rana esculenta

In the present study, we have localized for the first time Met- and Leu- enkephalin-like material in the ovary of the anuran, Rana esculenta, using the indirect immunofluorescence method. The ovaries were sampled during the main representative phases of the annual reproductive cycle of the frog, living in a mountain pond (Colfiorito, Umbria at 820 m a.s.l.). Strong immunoreactivity to Met- and Leu-enkephalin antisera was observed in the follicle cells of the granulosa layer of vitellogenic oocytes; moreover, during this phase, immunofluorescent materials were also radially localized in the cytoplasm and in the perinuclear zone. The mature oocytes showed Met- and Leu-enkephalin-like immunostaining in the thecal layer and in several granules scattered in the peripheral zone of the yolk. The different pattern of Leu- and Met-enkephalin-like immunoreactivity in the frog ovary parallels and complements the changes occurring in the reproductive (May) and in the vitellogenetic (September) phases during the ovarian cycle. Consequently, these findings strongly support the hypothesis for a local synthesis of these peptides in the ovary and suggest their possible involvement in the control of ovarian function.