Antiapoptotic and proliferative activity of curcumin on ovarian follicles in mice exposed to whole body ionizing radiation.

The aim of this study was to evaluate the antiapoptotic and proliferative activity of curcumin (Cur) on the ovarian follicles in mice exposed to whole body ionizing radiation (Rd). The mice were exposed to 8.3 gray whole body Rd, and Cur groups were given as a daily dose of 100 mg/kg of Cur for 10 days (10 days before Rd). The ovaries were collected 3 and 12 h after irradiation. To date, no such studies have been performed on antiapoptotic and proliferative activity of Cur on the ovarian follicles in mice exposed to whole body Rd. Analysis of mice ovary after exposure to Rd by terminal-deoxynucleotidyl-transferase-mediated dUTP nick end labeling showed that there were apoptotic cells both in the follicular wall and the antrum, and that the number of follicles showing early atresic features was high 3 h after Rd. On the other hand, analysis of mice ovary 12 h after exposure to Rd showed that the number of follicles containing apoptotic cells with advanced atresic features was significantly higher when compared to the 3-h Rd exposure group. The proliferating cell nuclear antigen -positive granulosa cells were decreased in association with follicular atresia. The groups given treatment were observed to have some benefit from Cur against the damage caused by Rd. The results of this study demonstrate that Cur prevents follicular atresia in Rd-induced apoptosis in ovarian follicles.

Gamma-radiation-induced follicular degeneration in the prepubertal mouse ovary.

Prepubertal mice were whole-body irradiated with a mean lethal dose (LD50) of gamma-radiation using a 60Co source with a total dose of 7.2 Gy and a dose rate of 12.0 cGy/min. At day 0 before the irradiation and at day 1, 2, and 3 after the irradiation, the ovaries were collected and the morphological changes were assessed. The ratios (%) of atretic or polymorphonuclear leukocytes (neutrophil)-infiltrated follicles in the largest cross sections were calculated. In the early atretic follicle of the control mouse ovary, both apoptotic and mitotic cells were observed and occasionally neutrophils were infiltrated into the follicle cavity. However, in the atretic follicles 2 days post-irradiation, numerous cell fragments, apoptotic cells and bodies, and especially, a number of neutrophils were observed. In the non-irradiated control, the ratios of atretic follicles were 58.0+/-8.6 and 27.3+/-11.2 (mean+/-S.E.M.) in antral and preantral follicles, respectively. The ratios of the number of antral and preantral follicles with one or more neutrophils to the total number of atretic follicles were 29.3+/-12.0. At 2 days post-irradiation, the ratios of atretic follicles were increased to 94.0+/-3.4 and 86.9+/-7.6 in antral and preantral follicles, respectively. The ratios of neutrophil-containing follicles among the atretic one were increased to 65.9+/-11.5 and 57.8+/-15.4 at 2 and 3 days after the irradiation, respectively. Taken together, the present results show that gamma-radiation induces apoptotic and inflammatory degeneration of mouse ovarian follicles. Besides, neutrophils may be involved in the acute atretic degeneration in gamma-irradiated mouse ovarian follicles.

Radiation-induced chromosome aberrations in guinea-pig growing oocytes, and their relation to follicular atresia.

The female guinea-pig has been shown to represent a good model to investigate the genetic hazard of ionizing radiation in humans. The sensitivity of the guinea-pig oocytes to radiation-induced chromosome aberrations was, therefore, studied at different stages of oocyte and follicular growth. The sensitivity of oocytes enclosed in small follicles (15 weeks before ovulation) was found to be low and comparable to that of immature oocytes present at birth. The sensitivity of growing oocytes remained low and almost constant until 3 weeks before ovulation, from which time it began to increase. The most dramatic increase of sensitivity occurred during the last week preceding ovulation: about 90% of oocytes X-irradiated with 4Gy, 2 days before ovulation showed one or more chromatid interchanges, as compared to 20% for those irradiated with the same dose 1 week earlier. A comparison of our results with those found by others in the mouse shows that considerable differences of sensitivity exist between oocytes of these two species irradiated at similar stages of development. The possible reasons for these differences are discussed.

[The dynamics of folliculogenesis in the ovary of the progeny of animals irradiated in embryogenesis and early postnatal life]. / Dinamika follikulogeneza v iaichnike potomstva ot obluchennykh v émbriogeneze i rannei postnatal'noi zhizni zhivotnykh.

The prolonged irradiation with 1.0 Gy dose in prenatal and early postnatal periods of development of rats leads to the disturbance in the rate of follicular growth, the reduction of their pool, mainly the pool of young forms of oocytes. Processes of degeneration and destruction occur and develop in the ovary of both the irradiated rats and their first progeny.

Decreased retinoblastoma protein expression in gamma-irradiated mouse ovarian follicles.

We immunohistochemically examined the effect of gamma-radiation on immature mouse ovarian follicles. Mice were Jirradiated with a dose of LD80 (8.3 Gy) in KAERI. At 0 h, 6 h, 12 h, 1 d, 2 d, 4 d and 8 d postirradiation, the ovaries were excised and fixed in neutral buffered formalin. We performed immunohistochemistry for protein retinoblastoma (pRb), terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) and routine hematoxylin-eosin staining in the largest cross sections. Radiation-induced follicular degeneration increased before 6 h, and most irradiated ovarian follicles became acutely atretic. The immunohistochemical staining for pRb was strong in the nuclei of granulosa cells of normal follicles and weak in atretic ones which were, conversely, strong for TUNEL staining. It was shown that pRb expression became lower with the degeneration of the ovarian follicles, which was inhibited by gamma-radiation. In the present study, pRb immunohistochemistry was proven to be a useful tool for the identification of follicular status.

Gamma-radiation accelerates ovarian follicular atresia in immature mice.

This study deals with the morphological degenerations of normal and atretic follicles based on artificially induced radiation apoptosis. ICR strain of 3 week-old female mice were whole body irradiated with 8.3 Gy and sacrificed by cervical dislocation. Ovaries were collected at 0 h, 6 h, 12 h, 1 d, 2 d, 4 d, and 8 d post irradiation and processed for morphological observation. Irradiated ovarian follicles showed such characteristics as faint connection between zona and cumulus thinning of granulosa layer, numerous apoptotic bodies, and increased cell debris in antrum. However, in normal ovaries about a half of the follicles were healthy. Within 6 h post irradiation, more than 95% of follicles steeply became apoptotic. At 4 d, severely damaged follicles with hypertrophed theca layer and fragmented oocytes were shown. At 8 d, normal shaped follicles were observed together with severely disrupted ones. The normal shaped follicles seemed to have grown from radioresistant ones. In conclusion, gamma-radiation had an additive or concomitant effect on atretic degeneration of mouse ovarian follicles.

Effect of exposure to continuous light and melatonin on ovarian follicular kinetics in the skipper frog, Rana cyanophlyctis.

Ovarian follicular kinetics and gravimetric changes in the ovary and oviducts were studied in the skipper frog, Rana cyanophlyctis, following exposure to continuous light and melatonin treatment during the breeding season. Daily late-afternoon injections of melatonin (15 micrograms subcutaneous) for 30 days decreased the gonadosomatic index (GSI), whereas continuously available melatonin from subcutaneous implants did not influence the GSI compared to those of controls. Exposure to continuous light for 30 days stimulated the GSI, and melatonin given as daily injections prevented the continuous-light induced increase in GSI. Oviductal weights decreased only in the melatonin-injected groups. Data on follicular kinetics revealed a decrease in first-growth-phase (FGP) oocytes and an increase in medium-sized second-growth-phase (MSGP) and large-sized second-growth-phase (LSGP) oocytes following continuous-light exposure. Melatonin administered to continuous-light-exposed frogs did not change the FGP oocyte number; however, it reduced both the MSGP and LSGP oocytes. Melatonin administration to frogs held in a light and dark cycle increased FGP oocytes and decreased MSGP and LSGP oocytes. Atretic follicles increased in all melatonin-treated groups. The results suggest that continuous light stimulates and melatonin inhibits reproductive function in this species.

An analysis of the effects of increasing doses of ionizing radiation to the exteriorized rat ovary on follicular development, atresia, and serum gonadotropin levels.

There is increasing interest in the effects of environmental and therapeutic agents on the reproductive system, in particular, the ovary. To study the effects of controlled doses of ionizing radiation to the ovary, Sprague-Dawley rats had their ovaries exteriorized and subjected to increasing doses of radiation. There was a significant increase in ovarian follicular atresia, a significant increase in serum follicle-stimulating hormone levels, but no change in serum luteinizing hormone levels. This experimental protocol may facilitate the testing putative radioprotectants.