Reaction of the ovaries of 1- and 4-day-old rat pups to transplantation

Morphofunctional peculiarities of ovarian development in 1- and 4-day-old rats have been studied under conditions of their transplantation into mature female rats. In 30 days in transplants of both series of the experiments atretic process develops. Better preservation of the general pool of the germ cells and follicles is obtained at transplantation of the ovaries from the 4-day-old donors; their ovocytes at the operation time are at the stationary state of meiosis prophase. Ovocytes of 1-day-old animals, being at the stages of early prophase, are more vulnerable at homotransplantation.     

Historical aspects of protandrous sex change in the anemonefish Amphiprion melanopus (Pomacentridae, Teleostei)

Gonadal structure and cellular composition were examined in juveniles, males and females of the protandric hermaphrodite, Amphiprion melanopus. Functional sex change was experimentally induced in the field and gonad structure was histologically examined both qualitatively and quantitatively at 10, 20, 30 and 45 days after its initiation. Juvenile gonads consist primarily of immature ovarian tissue. Functional male gonads are ovotestes with co-existing mature spermatogenic tissue and immature ovarian tissue, while females possess only ovarian tissue. The initiation of sex change is marked by a rapid maturation of spermatogenic tissue and proliferation of putative oogonia. Gonads were essentially female by 20 days into sex change, but evidence of mature female function (marked by the initiation of vitellogenesis) was not observed until 45 days. Considerable variation between individuals was seen in quantitative measures of gonadal change in the early stages of sex change, but not in later stages. Progress in sex change as indicated by histological indicators was, however, consistent within stages. Duet systems for gamete transport changed from the male to the female form after all male tissue had been replaced.     

Neonatal superior ovarian nerve transection disturbs the cyclic activity of the female rats

The neural pathway most related with ovarian steroidogenesis has been identified as the superior ovarian nerve (SON). This work constitutes the first study of the effects of early ovarian SON transection, which was performed in rats of 4 days of age (SON-t rats) to magnify the effects of the denervation. The rats were studied at the prepubertal (30 days), peripubertal (41 days) and adult cyclic in dioestrus (60 days) reproductive stages. The SON-t rats showed a delay of vaginal opening, a notable disruption of oestrous cyclicity, and a large number of corpora lutea. In all the stages, the circulating levels of FSH, prolactin and growth hormone were lower in SON-t rats than in controls, whereas LH did not vary. Serum androstenedione levels were higher in SON-t rats at 30 days and lower at 41 days, compared with control animals while no difference was observed at 60 days. Serum progesterone levels did not differ between control and SON-t, but serum oestradiol concentrations were higher in SON-t rats in all of the stages. At the peripubertal stage, there were fewer ovarian beta-adrenergic receptors in SON-t ovaries, associated with a rise in the ovarian content of norepinephrine, but no changes were observed in SON-t rats at 30 and 60 days with respect to the controls. The release of progesterone in vitro from luteal cell in SON-t rats at 60 days was reduced in basal condition and under ovine LH or FSH stimulation, when compared with control animals; while no difference was observed in presence of isoproterenol or androstenedione in the culture medium. In corpora lutea of SON-t rats at 60 days, no change was observed in the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), but the activity of 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) was reduced, suggesting abnormal luteolysis in spite of the large number of corpora lutea. The interruption of innervation at an early age by SON transection is very important in the regulation of ovarian development in prepubertal and cyclic rats. The functional changes observed in the ovary suggest a possible alteration in the hypothalamic-hypophyseal axis.     

Expression and localization of P-, K-, and OB-cadherin in the prepubertal rat ovary.

Classical and atypical cadherins mediate calcium-dependent cell adhesion and play an important role in morphogenetic processes. We have shown, previously, N- and E-cadherin expression in the rat ovary. This expression, however, was not associated with specific follicle-restructuring events such as antrum formation and segregation of mural from cumulus granulosa cells suggesting that other cadherins may serve this function. In this study, RT-PCR and immunostaining techniques showed that three other cadherins are expressed throughout prepubertal ovarian development in the rat: one classical (P-) cadherin, and two atypical (K- and OB-) cadherins. RT-PCR analysis of isolated ovarian tissue compartments (granulosa cells and the residual ovarian tissue) agreed with the immunostaining results. Immunostaining showed P- and K-cadherin expression by granulosa, as well as thecal/interstitial cells, and also in oocytes of primordial follicles. P-cadherin expression was absent in oocytes of follicles in later stages of development compared to K-cadherin, which was found in oocytes at all stages of folliculogenesis. P-, K-, and OB-cadherin were expressed by the ovarian surface epithelial cells of neonatal animals but only P- and OB-cadherin expression were maintained in these cells in 25 day-old animals. Cellular OB-cadherin staining was absent in follicles at all stages of development and its expression was restricted to the ovarian hilar region and portions of the stroma. In summary, cadherin expression and distribution profiles changed during ovarian growth and folliculogenesis suggesting a role for cadherins in organizational and morphogenetic processes within the developing rat ovary.     

The relationship of age and ovarian development to mating of Delia antiqua (Diptera: Anthomyiidae)

Abstract. The relationship between age and sexual receptivity for male and female onion flies, Delia antiqua (Meigen) (Diptera: Anthomyiidae), and factors affecting ovarian development were examined under various conditions. Confinement in small arenas had a significant negative effect on the rate of oogenesis over the first 10 days of adult life and, in some cases, on the percentage of females inseminated. The rate of oogenesis was not affected by male population density, but high density of females in large arenas appeared to have a priming effect. Few males in single-pair mating bioassays conducted over 24 h were sexually mature at 3–4 days post-eclosion, but > 50% mated when aged 6–7 days. Although females first mated at 3–4 days post-eclosion when confined for 24 h with sexually mature males, most females mated at 6–7 days of age when oocytes in the terminal ovariole position comprised 50% of total egg volume (stage 7 or greater on a 10-stage scale). No females aged 3 days were mated in no-choice and two-choice bioassays with sexually mature males over 24 h. Previtellogenic females (stage 3) were not inseminated, and ovarian development was only correlated weakly ( r = 0.48) with mating. Frequency of mating in mixed-sex groups of twenty flies was comparable with that in single pairs. However, more females were inseminated at 3–4 days, probably as a result of multiple matings by a few precocious males. These results do not support the hypothesis that females mate only when ovarioles are mature.     

Mouse oocytes derived from fetal germ cells are competent to support the development of embryos by in vitro fertilization

Mouse oocyte development in vitro has been studied in the past several years, but no evidence showed that the fertilizable oocytes could be obtained from the fetal mouse germ cells before the formation of the primordial follicles. In this study, an efficient and simple method has been established to obtain the mature oocytes from the fetal mouse germ cells at 16.5 days post-coitum (dpc). For the initial of follicular formation, fetal mouse 16.5 dpc ovaries were transplanted to the recipient under the kidney capsule, and the ovaries were recovered after 14 days. Subsequently, the growing preantral follicles in the ovarian grafts were isolated and cultured in vitro for 12 days. Practically, the mature oocytes ovulated from the antral follicles were able to be fertilized in vitro and support the embryonic development. The results demonstrate that the fetal mouse 16.5 dpc germ cells are able to form primordial follicles with the ovarian pregranulosa cells during the period of transplantation in the ectopic site, and the oocytes within the growing follicles are able to mature in vitro, then are able to support the embryonic development.     

8-hour rhythm of ovarian progesterone secretion in ewe

Pulse character of hormones secretion in the hypothalamus-pituitary-gonads system is a necessary condition of physiological regulation of reproduction. At the same time, the rhythms of ovarian hormones secretion have not been adequately explored. The researches study mainly three sexually mature ewes. The stages of oestrus cycle were determined on behavioral reactions of females in the presence of ram. Blood samples from jugular vein were collected hourly over 24-hour period during follicular (15-16 days), early (3-4 days) and middle (7-9 days) luteal phase of oestrus cycle, pregnancy (40-105 days) and lactation (30-45 days). 27 experiments were performed. Plasma progesterone was determined by enzyme-immunoassay method. There was no diurnal rhythm of ovarian progesterone secretion in ewes. During early and middle luteal phase of oestrus cycle and lactation, an 8-hour rhythm of progesterone secretion was detected. Follicular phase of oestrus cycle and pregnancy were characterized by irregular rises of fluctuations of progesterone level. It seems that the 8-hour rhythm of progesterone secretion during luteal phase and lactation is controlled by action of intraovarian generator of ultradian rhythms.     

Orthotopic Transplantation of Cryopreserved Mouse Ovaries and Gonadotrophin Releasing Hormone Analogues in the Restoration of Function following Chemotherapy-Induced Ovarian Damage

Therapy advances are constantly improving survival rates of cancer patients, however the toxic effects of chemotherapy drugs can seriously affect patients’ quality of life. In women, fertility and premature ovarian endocrine dysfunction are of particular concern. It is urgently we find methods to preserve or reconstruct ovarian function for these women. This study compares GnRHa treatment with ovarian tissue cryopreservation and orthotopic transplantation in a chemotherapy-induced ovarian damage murine model. 56 inbred Lewis rats were divided into 4 treatment groups: Saline control (group I); cyclophosphamide only (group II); cyclophosphamide plus GnRHa (group III); cyclophosphamide and grafting of thawed cryopreserved ovaries (group IV). Body weight, estrous cycle recovery time, ovarian weight, morphology and follicle count, as well as breeding and fertility were compared among groups. Only group IV was able to restore to normal body weight by the end of the observation period and resumed normal estrous cycles in a shorter time compared to other treatment groups. There was a decrease in primordial follicles in all treatment groups, but group III had the greatest reduction. Although, there was no difference in pregnancy, only one animal littered normal pups in group II, none littered in group III and four littered in group IV. Thus, cryopreservation and orthotopic transplantation of ovarian tissue can restore the fertility of rats subjected to chemotherapy in a manner that is superior to GnRHa treatment. We also observed increased rates of hepatic, splenic and pulmonary haemorrhage in group III, suggesting there may be synergistic toxicity of GnRHa and cyclophosphamide.     

Gonadotropin secretion and testicular function in golden hamsters exposed to skeleton photoperiods with ultrashort light pulses

Both sexually mature and sexually regressed male golden hamsters were transferred to asymmetric skeleton photoperiods with night interruptions of varying duration, the short pulses occurring 14 h after "dawn." Testicular function and accompanying changes in follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone and spermatogenesis were observed. Sexually regressed animals exposed to a night-break of 6 seconds (sec) or longer exhibited maximal testicular development with a rapid rise in FSH secretion followed by a slower, more variable rise in LH. Full testicular size was achieved after 8 weeks. Night-breaks of 250 milliseconds (msec) or 1 sec induced testicular development and spermatogenesis but at a slower rate: levels of FSH and LH were still rising at the end of the experiment. Complete testicular maintenance was achieved by night-breaks of 1 sec or longer. Partial testicular regression was observed with a night-break of 250 msec. Night-breaks (60 sec) given less frequently than daily also stimulated testicular function and a night-break every 7 days increased FSH and LH secretion in sexually regressed hamsters, causing testicular development at a submaximal rate. Night-breaks given more frequently induced rapid testicular growth. Almost complete testicular maintenance of sexually mature hamsters was achieved with a 60-sec night-break at weekly intervals. Symmetric skeleton photoperiods also triggered testicular development in sexually regressed hamsters, with two 1-sec light pulses (14 h apart) being almost as effective as a normal long day. No difference in reproductive function was observed between animals on long days (14L:10D) and those exposed to maximally stimulatory skeleton photoperiods.     

Development of antral follicles in cryopreserved cat ovarian tissue transplanted to immunodeficient mice

Ovarian cortex cryopreservation and xenotransplantation into immunodeficient mice represents a potential means for female germplasm conservation and an immediate model for investigation of folliculogenesis. The objectives of this study were to: (1) assess follicle survival after cryopreservation and transplantation of cat ovarian tissue into non-obese diabetic severely combined immunodeficient (NOD SCID) mice; and (2) evaluate the effects of gonadotropin treatments on follicular development in the transplanted tissue. Slices from the cat ovarian cortex were frozen and after thawing, transplanted under each kidney capsule of castrated male NOD SCID mice (eight xenografts in four mice). Sixty-two days after surgery, mice were randomly assigned (two per group) to gonadotropin-treated (eCG and hCG 88 h later) or control (saline-treated) groups. Twenty-four hours after the last injection, ovarian tissue was recovered and processed for histology. Fresh ovarian tissue from the same original source was similarly processed. Follicles were counted, measured, and classified as primordial, primary, secondary, or antral. Immunoreactive proliferating cell nuclear antigen (PCNA) stain was used to assess follicle viability. Microscopic examination revealed no evidence of necrosis or fibrosis. The grafts were well-vascularized, with follicles at all stages of development. Numbers of follicles in the transplanted tissue were markedly reduced compared to fresh tissue, with approximately 10% of follicles surviving freezing and transplantation procedures. Growing follicles positive for PCNA were found in all xenografts. Gonadotropin treatment did not alter the proportion of resting to growing follicles or mean follicle diameter by comparison with controls from untreated mice. By contrast, luteinization, but not ovulation, of antral follicles was observed only in grafts from treated mice. In summary, frozen-thawed cat ovarian cortex tissue not only survived xenotransplantation, it also contained follicles able to grow to antral stages. Exogenous gonadotropin treatment in this model resulted in luteinization of antral follicles but enhancement of follicular growth and ovulation did not occur.     

Cysts and tumour-like lesions in the endocrine pancreas of the lamprey (Lampetra fluviatilis)

Histological observations on the endocrine pancreas of the upstream migrant stage of the River lamprey, Lamperra fluviatilis , have revealed a relatively high incidence of cysts and other tumour-like lesions, remarkably similar to the hamartomas described in the Atlantic hagfish, Myxine glufinosa. The frequency of these lesions increases throughout the migratory period from autumn to spring. In the most common type the cysts are surrounded by an intensely granular, single-layered epithelium and both the epithelial cells and the cyst contents show a marked reaction to aldehyde-fuchsin staining. In more extreme forms the breakdown of the epithelial walls of neighbouring cysts leads to the formation of large multi-locular structures occupying almost the whole of the islet organ, but no consistent evidence of changes in blood sugar levels has been obtained in animals where the islet tissue is severely affected. A smaller proportion of the animals display neoplastic developments, frequently associated with hyperplasia and vacuolization of the foregut epithelium. In such cases, there is evidence of invasiveness and the development of epithelioid nodules within the adjoining islet tissue. Cavitation within these nodules leads to the formation of cyst-like structures containing cell debris which reacts to aldehyde-fuchsin staining. The development of cysts and hyperplasia is interpreted as a renewal in the sexually mature lamprey of inherent trends towards epithelial proliferation and vacuolization that are involved during metamorphosis in the morphogenetic processes that lead to the formation of the adult foregut and cranial islet tissue.     

实验卵巢瘤肿的形态研究

瘤肿发生的原因至今还不甚明瞭,但几十年东由于在动物方面的研究以及临床方面的观察知其原因是多方面的,例如:病毒,物理的和化学的刺激,遗传的因素等等,对某些瘤肿的发生来说,都有重要的系关。实验证明,某些激素在一定的情况下也能诱导瘤肿的发生。动情素有刺激组织生长的能力,所以在它的长期作用下,很多器官例如:乳腺,子宫,脑垂腺等,都能从善性增生的基础上形成瘤肿。Burrows and Horning(1952)和 Lipschutz(1950)的专著对固醇激素,特别是动情素,与瘤肿的系关有了详尽的讨论。同样,脑垂腺激素也有类似的作用。近年来某些品系的小鼠在阉割后可发生肾上腺皮质瘤的发现[Woolley 和 Little(1945)],和抗     

In utero exposure of mice to diethylstilbestrol alters neonatal ovarian follicle growth and development

The prenatal exposure of mice to diethylstilbestrol (DES, 10 micrograms/kg on day 15 of gestation) caused both quantitative and structural alterations in ovarian follicles within the neonatal ovary. At birth, control ovaries consisted of small type 1 and 2 ovarian follicles located in the ovarian cortex. By postnatal day 7, ovarian follicle development had advanced to the type 4 stage with larger follicles located within the ovarian medulla. In DES-exposed animals, ovarian follicle maturation was advanced with type 3b and 4 follicles appearing 24 h prior to their appearance in control animals. Also, type 5 ovarian follicles were present on postnatal day 6 in experimental animals but were never seen in control animals. In addition to an alteration in ovarian follicle dynamics, the diameter of individual ovarian follicles was transit time between the various stages of follicular development which results in a greater number of developmentally advanced ovarian follicles being present during neonatal ovarian development. The mechanism by which prenatal exposure to DES alters ovarian follicle dynamics during neonatal development is not known.     

Müllerian-inhibiting substance inhibits rat Leydig cell regeneration after ethylene dimethanesulphonate ablation

The postnatal development of Leydig cell precursors is postulated to be controlled by Sertoli cell secreted factors, which may have a determinative influence on Leydig cell number and function in sexually mature animals. One such hormone, Mullerian inhibiting substance (MIS), has been shown to inhibit DNA synthesis and steroidogenesis in primary Leydig cells and Leydig cell tumor lines. To further delineate the effects of MIS on Leydig cell proliferation and steroidogenesis, we employed the established ethylene dimethanesulphonate (EDS) model of Leydig cell regeneration. Following EDS ablation of differentiated Leydig cells in young adult rats, recombinant MIS or vehicle was delivered by intratesticular injection for 4 days (Days 11-14 after EDS). On Days 15 and 35 after EDS (1 and 21 days post-MIS injections), endocrine function was assessed and testes were collected for stereology, immunohistochemistry, and assessment of proliferation and steroidogenesis. Although serum testosterone and luteinizing hormone (LH) were no different, intratesticular testosterone was higher on Day 35 in MIS-treated animals. At both time points, intratesticular 5alpha-androstan-3alpha,17beta-diol concentrations were much higher than that of testosterone. MIS-treated animals had fewer mesenchymal precursors on Day 15 and fewer differentiated Leydig cells on Day 35 with decreased numbers of BrdU+ nuclei. Apoptotic interstitial cells were observed only in the MIS-treated testes, not in the vehicle-treated group on Day 15. These data suggest that MIS inhibits regeneration of Leydig cells in EDS-treated rats by enhancing apoptotic cell death as well as by decreasing proliferative capacity.     

Ovarian development in the Amazonian red discus, Symphysodon discus Heckel (Osteichthyes: Cichlidae).

The Neotropical red discus fish, Symphysodon discus, originates from the Amazonian basin and has a discoid body. Although this species is popularly used in aquaria and is exported as an ornamental fish, it has not been intensively studied. The purpose of this work was to study the morphological and histological aspects of the ovarian development in the red discus. Forty females of S. discus of varying body sizes and different stages of gonadal development were used in this study. The ovaries were weighed and examined macroscopically in order to observe the maturation stages. Histological staining of hematoxilin-eosin was used for microscopic observations of the ovaries. Macroscopic observations showed immature, maturing, mature and partially spent stages of ovarian development. Microscopic examination of the ovaries revealed five stages and five phases of ovarian development. The ovaries of the mature females showed all the different phases of oocyte development, indicating the multiple spawning habit of this species.     

Effect of mesotocin on thyroid gland of the frogRana temporaria

The thyroid gland was studied in intact and hypophysectomized (10 and 60 days before the experiment), sexually mature male frogsRana temporaria after three injections of mesotocin at 30-min intervals (a dose of each injection was 5 ng/g body weight). As determined morphologically, height of the thyrocytes was statistically significantly decreased in intact frogs, while no statistically significant changed were revealed in hypophysectomized animals. The data obtained indicate that at stress responses in frogs, mesotocin can affect the thyroid gland via adenopituitary rather than via general circulation, i.e., by an indirect way.     

Cryoconservation des tissus testiculaires: Quel avenir?

Over the last twenty years, an increased incidence of cancer has been observed in the population of adolescents and young adults. With the progress in cancer diagnosis and therapy, childhood cancer has become a curable disease. The efficacy of treatment is associated with a high degree of toxicity and gonadal function is particularly sensitive to this toxicity. Prevention of sterility in childhood cancer survivors will become a major challenge in reproductive medicine. Cryopreservation of ovarian tissue is performed in girls and women before cancer treatment. Cryopreservation of ejaculated spermatozoa is possible in sexually mature boys. However, for prepubertal boys or after failure of ejaculated sperm collection, mature or immature testicular tissue banking should be proposed. However, an optimal cryopreservation protocol is a prerequisite for clinical applications. Furthermore, the future applications of immature testicular tissue banking should be developed, not solely autologous testicular tissue grafting, but also in vitro maturation of germ cells. Cryopreservation protocols, transplantation and in vitro maturation techniques should be improved in animal models and in humans.     

Ovarian activity and uterus organometry in delayed puberty gilts

About 30% of the total number of gilts selected for reproduction at the large breeding farm units in Vojvodina (Republic of Serbia) are culled due to prolonged pre-insemination anoestrus (estrus not detected until 8 mo of age). The aim of this study was to provide the answer to the following question: do the culling gilts reach cyclic ovarian activity at all? One hundred seventy five culled gilts in which external estrus manifestations were not detected by 8 mo of age were sacrificed and their reproductive organs were examined for determination of sexual maturity (ovaries exhibiting pre-ovulatory follicles 8 to 11 mm in diameter, corpora hemorrhagica, corpora lutea and corpora albicantia). Uterine weights and horn length were also determined. Functional ovaries were observed in 107 (61.1%) examined gilts, with 62 animals having one and 45 having two puberty ovarian cycles (57.9% and 42.1%, respectively). Pathomorphological changes which could result in prolonged pre-insemination anoestrus were not observed on the reproductive organs of sexually mature gilts. Our results indicate that most of the culling gilts have reached cyclic ovarian activity. The main reason for culling due to the absence of external estrus manifestations in sexually mature gilts could be inadequate estrus detection technology.     

Effects of pregnant mare serum gonadotropin (eCG) on follicle development and granulosa-cell apoptosis in the pig

The effect of eCG on follicular development and granulosa-cell apoptosis in sexually mature and immature gilts and on granulosa-cell apoptosis in vitro were studied. The sexually mature gilts were treated with eCG on Day 11 of the estrous cycle, and effects were analyzed at different times after treatment with untreated animals at corresponding stages of the cycle as controls. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), hematoxylin and eosin staining, and DNA ladder. The proportion of apoptotic cells in atretic follicles (39%) was significantly higher (P<0.01) than that in healthy follicles (9%). At 24h after eCG treatment in mature gilts, the total number of follicles visible on the ovarian surface (57 per ovary), the number of small (<3mm) follicles (31.5 per ovary) and the number of medium-sized (3-5mm) follicles (23 per ovary) were significantly higher (P<0.05) than those of control animals (28, 20 and 6.5 per ovary, respectively), and declined gradually thereafter to below the level of control animals. The number of large (>or=5mm) follicles began to show a marked increase at 72h after eCG (8.5 versus 2.5, P<0.05). At 24h after eCG treatment, the proportions of apoptotic cells in small (7.2%) and medium-sized follicles (7.4%) were markedly lower (P<0.01) than those in controls (21.5 and 21%, respectively) and increased gradually thereafter to approach the level in controls. The percentage of apoptotic cells in large follicles (10% at 24h post-eCG) did not change significantly. Before eCG treatment, there were markedly fewer follicles of all types on ovaries of immature gilts than of mature gilts (9 versus 25 per ovary) and the proportion of apoptotic cells in small and medium follicles was high (25 and 34%, respectively). After eCG treatment, the changes in follicle number and proportion of apoptotic cells in the immature gilts followed a similar pattern to that of the mature gilts. Equine chorion gonadotropin inhibited apoptosis of granulosa cells cultured either in vitro or in intact follicles in a dose-dependent manner. Thus, follicular atresia in the pig, as in other animals, was characterized by apoptosis of large numbers of granulosa cells, and eCG promoted follicular development by inhibition of granulosa-cell apoptosis.     

The development of carp gonads in warm water aquaria

The development of carp gonads in warm water (23°C) aquaria under controlled conditions is described. In this environment males mature at 6 months of age. Females ripen at 14 months, and by 15 months 25% of these have started to spawn, whereas under natural conditions female European carp do not ripen until they are 4 years old. Fishes do not lay all their eggs at once, as evinced by the finding of partly-spent ovaries. Sexual maturation of the ovaries has been described in six stages, based on histological changes, and associated with changes in the female gonado-somatic index and the maximum ova diameter. The maturation of testes occurs in four stages. The percentage incidence of each stage of ovarian maturation has been calculated for the 6th to the 15th month. Larger fishes become sexually mature before smaller fishes.