The morphology of healthy and atretic follicles in the ovary of the sexually immature ostrich was described in the present study. In addition, the distribution of the intermediate filaments desmin, vimentin and smooth muscle actin, in these ovarian follicles, was demonstrated. Healthy and atretic primordial, pre-vitellogenic and vitellogenic follicles were present in the ovaries of the sexually immature ostrich. Atresia occurred during all stages of follicular development. Atretic primordial and pre-vitellogenic follicles were characterized by the presence of a shrunken oocyte surrounded by a multilayered granulosa cell layer. Two forms of atresia (types 1 and 2) were identified in vitellogenic follicles. In the advanced stages of type 1 atresia the follicle was dominated by a hyalinized mass. In contrast, in type 2 atresia the granulosa and theca interna cells differentiated into interstitial gland cells. Positive immunostaining for desmin was observed in the granulosa cells of only healthy primordial and pre-vitellogenic follicles. Atretic primordial and pre-vitellogenic follicles were immunonegative for desmin. Vimentin immunoreactivity was demonstrated in the granulosa cells of all follicles except the vitellogenic atretic follicles. The results of the present study indicate that ovarian follicles in the sexually immature ostrich undergo a cycle of growth and regression, which is similar to that reported in other avian species. Furthermore, based on the results of the immunohistochemical study, it would appear that the distribution and immunostaining of intermediate filaments changes during follicular development and atresia.

Keywords:Di-(n-butyl) phthalate Leydig cell Steroidogenesis Male Japanese quails Endocrine disruptionIn the present study, we have investigated the effects of 30-day dietary (pre-pubertal) exposure to different doses (0 (control), 1, 10, 50, 200 and 400 mg/kg bodyweight/day) of di(n-butyl) phthalate (DBP) on Leydig cells of adult male Japanese quails by quantifying the transcript levels for P450 side-chain cleavage (p450scc), P450c17 (CYP17), and 3β-and 17β-hydroxysteroid dehydrogenase (hsd) using quantitative (real-time) poly-merase chain reaction (qRT-PCR). In addition, the plasma testosterone levels were analysed using radioimmuno-assay (RIA) and testis was examined for evidence of gross pathology and histopathology. Our data showed that pre-pubertal exposure to DBP produced alterations in testicular architecture as evident by poorly developed or misshaped testis, and altered spermatogenesis due to tubular degeneration and atrophy of seminiferous tubules especially in the high DBP dose (200 and 400 mg/ kg) treated groups. In addition, DBP altered several key en-zymes involved in testicular steroidogenesis pathways in an apparent dose-dependent manner. For example, bi-phasic effects of DBP were observed for P450scc and 3β-hsd mRNA, that were generally increasing at low dose 10 mg/kg, and thereafter, an apparent dose-dependent decrease between 50 and 400 mg/kg. The steroidogenic acute regulatory (StAR) protein was at the lowest detectable limits and therefore not quantifiable. These effects did not parallel the non-significant changes observed for plasma testosterone levels. The present data is consis-tent with previous reports showing that DBP modulates Leydig cell steroidogenesis in several species, with a po-tential negative effect on reproduction in those avian species that are vulnerable to endocrine disrupting chemicals.

The testicular capsule and peritubular boundary tissue of the emu and ostrich, as typical representatives of ratite birds, were studied in sexually mature and active birds. The testicular capsule was much thicker (578.1±73.4 m for the free surface of the ostrich testis, and 176.2±57.5 mfor the emu) than those of members of the Galloanserae. The cellular composition of both testicular capsule and peritubular tissue was similar generally to that of members of the previously studied Galloanserae and of mammals. The tunica albuginea of the testicular capsule mainly comprised smooth-musclelike or myoid cells mostly running in one direction and occurring in one main mass. Unlike the Galloanserae, the tunica albuginea contained more collagen fibres than smooth muscle cells, especially in the ostrich. Peritubular tissue was similarly composed of smooth-muscle-like cells distributed in several layers. Actin microfilaments and desmin and vimentin intermediate filaments were variably immunoexpressed in these two tissue types in both birds, with a clear dichotomy in the peritubular tissue. Thus, taken together with studies of some members of the Galloanserae, avian testes clearly contain a morphological mechanism that is represented partly by the smooth muscle cells of the testicular capsule and peritubular tissue for transporting the testicular fluid, which is usually copious in birds, and its cellular content from the testis into the excurrent duct system; thismechanism is similar to that found in mammals.

SummaryThis study describes the distribution of vimentin, desmin, smooth muscle actin (SMA) and laminin in the oviduct of the laying domestic fowl. Vimentin immunostaining was The results of the study indicate that the immunolocalization of desmin, SMA and laminin in the oviduct of the domestic fowl is similar to that in the mammalian uterus.The immunolocalization of vimentin in the domestic fowl varies depending on the oviductal region.

The immunohistochemical localization of the intermediate filaments desmin, vimentin and smooth muscle actin (SMA) in the ovary of the emu was described in the present study. The cortical region of the ovary contained developing and atretic primordial, pre-vitellogenic and vitellogenic follicles. Vimentin immunostaining was demonstrated in the granulosa cell layer of primordial, pre-vitellogenic and vitellogenic developing and atretic follicles. An interesting finding of the present study was the localization of SMA in fibroblasts located in the theca externa of late vitellogenic follicles. The presence of SMA in these fibroblasts suggests that they possess characteristics of smooth muscle cells.

The innervation of the ovary has been studied in various species of birds and mammals. Despite the fact that the innervation of any organ is an essential factor in controlling its growth and function, no information is available on the distribution of nerve fibers in the ovary of the sexually immature ostrich. Thus, the present study was undertaken to investigate the distribution of nerve fibers in the ovary of the sexually immature ostrich, using antibodies against neurofilament protein type M of 160 kD (NP), protein gene product 9.5 (PGP 9.5) and neuron specific enolase (NSE). A total of 26 sexually immature female ostriches, aged between 12 and 14 months were used in the present study. Immunostaining was performed using a LSAB plus kit (Dakocytomation, Denmark). Antibodies against NP and PGP 9.5 were used at dilutions of 1:25 and 1:50, respectively. A ready-to-use solution containing antibodies against NSE was also used. Strong immunostaining for NP, PGP 9.5 and NSE was observed in nerve bundles, which coursed through the ovarian stalk and extended into the medulla and cortex. In addition, NSE immunoreactive nerve cell bodies were observed in the cortex and medulla. NP, PGP 9.5 and NSE immunoreactive nerve fibers were present in the thecal layer of the follicular wall. The current study has highlighted the distribution of NP, PGP 9.5 and NSE-immunoreactive nerve fibers in the ovary of the sexually immature ostrich. The findings of the present study suggest that the distribution of nerve fibers in the immature ostrich is similar to that of the domestic fowl.

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The present study investigated the distribution of nerves in the ovary of the emu. The neuronal markers, protein gene product 9.5, neurofilament protein and neuron specific enolase demonstrated the constituents of the extrinsic and intrinsic ovarian neural systems. The extrinsic neural system was composed of ganglia in the ovarian stalk, as well as nerve bundles, which were distributed throughout the ovary. Isolated neuronal cell bodies, in the medulla and cortex, formed the intrinsic neural system. An interesting finding of the study was the presence of nerve bundles, circumscribed by lymphocytes, in the ovarian stalk. The findings of the study indicate that the distribution of nerve fibres and neuronal cell bodies in the emu ovary is similar, but not identical to that of the domestic fowl and ostrich.