Morphological relationships and leukocyte influence on steroid production in the epigonal organ-ovary complex of the skate, Leucoraja erinacea.

In elasmobranchs, a unique association exists between an immune tissue, the epigonal organ (EO), and the gonads. In this study, the histological and vascular relationships of the EO and ovarian follicles of the little skate, Leucoraja erinacea, were assessed. Perfusions of Evans blue dye and Batson's monomer showed a shared vascular pathway from the gonadal artery into the epigonal-ovary complex, with blood first entering the EO and then perfusing the ovarian follicles. Histological studies demonstrated direct cellular contact between epigonal leukocytes and the follicle wall (FW), as well as the presence of leukocytes between the steroidogenic theca and granulosa cells. In vitro analyses demonstrated that epigonal cells co-cultured with FW cells cause a dose-dependent inhibition of estrogen (E2) and testosterone (T) production. In contrast, conditioned media from epigonal leukocytes, stimulated or unstimulated with lipopolysaccharide (10 microg/ml), increase the production of E2 and T from FW cells of the ovaries. These studies provide a basis for further investigations of leukocyte secreted factors and cell contact modulation of follicular steroid production.

Influence of reproductive activity, sex steroids, and seasonality on epigonal organ cellular proliferation in the skate (Leucoraja erinacea).

In elasmobranchs, the epigonal organ, a unique leukopoietic immune tissue, is associated with the gonads. As the ovaries increase in size during reproductive activity, the overall mass of the epigonal organ does not change. However, immunohistochemistry (proliferating cell nuclear antigen Ab) demonstrated more proliferative activity and extravasation of epigonal leukocytes from blood vessels in reproductively active (RA) skates (Leucoraja erinacea) than in non-reproductively active (NRA) skates. In addition, [(3)H]thymidine incorporation was greater in epigonal leukocytes from RA skates than in leukocytes from NRA skates. Plasma from RA skates, but not from NRA skates, increased proliferation of epigonal leukocytes in vitro, an effect that was not seen using steroid-free plasma. In contrast to the stimulatory effect of plasma on leukocyte proliferation, addition of steroids (estrogen, progesterone, testosterone, and dexamethasone) in vitro decreased [(3)H]thymidine incorporation. While the inhibitory response to steroids was seasonally variable, (3)[H]thymidine incorporation was always highest in RA animals, in which plasma steroid levels were also consistently highest. These studies suggest functional interactions between reproductive and immune tissues in the skate, and that cellular turnover in epigonal tissue may be influenced by gonadal activity.

Effects of reproductive activity and sex hormones on apoptosis in the epigonal organ of the skate (Leucoraja erinacea).

In elasmobranchs, a unique association exists between an immune tissue, the epigonal organ, and the gonads. The intimate morphological relationship between these tissues suggests functional interactions. In this study, we used apoptosis to assess differences between epigonal tissues of reproductively active (RA) and non-reproductively active (NRA) skates (Leucoraja erinacea). Plasma steroid levels were significantly higher in RA than in NRA animals, and TUNEL analysis showed that epigonal tissue of RA skates had greater DNA fragmentation than NRA skates. Addition of steroids to epigonal leukocytes in vitro demonstrated that progesterone, testosterone, and dexamethasone, but not estrogen, induced apoptosis of epigonal leukocytes as evidenced by DNA laddering and caspase-3 antibody labeling. This study supports recent evidence that cellular homeostasis of epigonal lymphomyeloid tissue may be influenced by gonadal activity and reproductive steroids in a representative of the most basal gnathastome group.