哺乳动物卵巢中生殖干细胞的研究历史与进展

对于出生后的哺乳动物卵巢中是否存在生殖干细胞以维持卵泡的更新一直争议不断,现就该领域的研究历史及最新进展进行综述.     

New strategy for in vitro activation of primordial follicles with mTOR and PI3K stimulators

It had been known for decades that primordial follicles in mammalian ovaries are assembled with definite numbers and represent the ovarian reserve throughout the reproductive life. Intra-oocyte PI3K/mTOR pathways have been indicated to play a central role on the activation of primordial follicles. Genetic modified mouse models with chronic activation of PI3K/mTOR signals in primordial oocytes showed premature activation of all primordial follicles and eventually their exhaustion. On the other hand, this may suggest that, unlike chronic activation of PI3K/mTOR, its acute activation in infertility would activate primordial follicles, permitting fertility during the treatment. Previously, PI3K stimulators were reported as a temporary measure to accelerate primordial follicle activation and follicular development in both mouse and human, and were applied in the treatment of infertility in premature ovarian failure (POF) patients. To address whether mTOR stimulators could play similar role in the process, we transiently treated neonatal and aged mouse ovaries with mTOR stimulators-phosphatidic acid (PA) and propranolol. Our results demonstrated the stimulators increased activation of primordial follicles and the production of progeny. Human ovarian cortex cubes were also treated with mTOR or/and PI3K stimulators in vitro. When they were used separately, both of them showed similar promotive effects on primordial follicles. Surprisingly, after joint-treatment with the 2 kinds of stimulators together, synergistic effects on follicular development were observed. Based on increased efficiency of follicular activation in humans, here we propose in vitro transient treatment with mTOR and PI3K stimulators as an optimized protocol for the application in different clinical conditions with limited follicle reserve.     

A review of ovarian follicular cysts in cows, with comparisons to the condition in women, rats and rabbits

Some morphological and endocrinological aspects of cystic follicles in cows, women, rats, and rabbits were reviewed. The etiology of follicular cysts remains a mystery, in part due to the difficulty of studying them during formation, particularly in women and cows. Investigations of the morphological and endocrinological nature of cystic follicles during their formation should enhance our understanding of the factors that cause them. Rats and rabbits may provide useful and convenient experimental models for the study of factors which contribute toward cyst formation.     

Association of laminin with heparan and chondroitin sulfate-bearing proteoglycans in neurite-promoting factor complexes from rat schwannoma cells

The present studies were undertaken to confirm the presence and identity of a putative proteoglycan associated with laminin in neurite-promoting factor complexes isolated from rat schwannoma cell conditioned medium. Sucrose density gradient centrifugation of the complex resolved two laminin-associated Na₂[³⁵S]O₄-labeled peaks which were termed Pools A and B. Both pools had nearly all their [³⁵S] cpms associated with glycosaminoglycan, contained heparan sulfate-proteoglycan core protein antigen and displayed a similarly high neurite promoting potency relative to their laminin contents. However, Pool A contained about twice as many [³⁵S] cpms and twice as much proteoglycan core protein per laminin than Pool B. Seventy percent of Pool A cpms was associated with heparan sulfate and 30% with chondroitin sulfate whereas the inverse was true for Pool B. Treatment with heparitinase and/or chondroitinase ABC caused laminin in either pool to elute at lower salt concentrations from DEAE cellulose. In SDS-PAGE the [³⁵S] cpms of both pools ran with the same mobility as laminin but could be separated from laminin under reducing conditions. The Pool A cpms remained at 900 KD and the Pool B cpms spread over the 200–900 KD range. By rotary shadowing electron microscopy, Pool B fractions contained primarily cross-shaped laminin images, often associated with proteoglycan-like images. Pool A fractions contained i) dense, aggregated images including intact laminin from which emanated proteoglycan-like strands, ii) circular images bearing globular domains and less commonly, iii) distorted cross-shaped laminin-like images. These studies support the existence of at least two forms of laminin-proteoglycan complexes which differ in biochemical, immunochemical and ultrastructural characteristics.Abbreviations HSPG heparan sulfate proteoglycan - ELISA enzyme-linked immunoassay - Pool A Fractions 10–13 in sucrose gradient (Figure 1, lower panel) - Pool B Fractions 14–16 in sucrose gradient (Figure 1, lower panel) - HDPG High density proteoglycan, Fractions 1–3 in cesium chloride gradient, Figure 1, middle panel - CPC cetylpyridinium chloride - GAG glycosaminoglycan Special issue dedicated to Dr. E. M. Shooter and Dr. S. Varon.     

Induction of atresia of the dominant follicle in rhesus monkeys (Macaca mulatta) by the local application of estradiol-17β

Estradiol-17β (E₂), administered systemically to rhesus monkeys during the follicular phase of the menstrual cycle, induces atretic changes in the microenvironment of the dominant follicle (DF), which results in its demise. It has been proposed that this effect of E₂ represents a direct action at the ovarian level. The present study was designed to test this hypothesis, using local treatment with E₂. After identification of the DF during laparoscopy on day 6 of the cycle, female monkeys were laparotomized and their ovaries exposed. Either corn oil (20 μl, controls) or E2 (100 μg ) in oil vehicle (experimentals) was injected into the ovary near the DF. In control animals, preovulatory release of gonadotropins and ovulation were normal in five of six animals, with cycle and luteal phase lengths of 27.8 ± 2.2 days and 14.6 ± 2.5 days, respectively (x? ± S.D.). Conversely, in only one of six animals in the experimental group did ovulation occur at the expected time (P < 0.05). In the other five treated animals, E₂ induced atresia of the DF and significantly extended cycles (35.4 ± 5.4 days) without affecting luteal phase lengths (12.0 ± 1.4 days). Concentrations of estrogen in peripheral sera of some animals were increased transiently at 6 h after injection of E₂ but returned to normal by 12 h; this duration of estrogen exposure is far less than the 24 h required to induce atresia of the DF in previous studies. At 6 h after injection of E₂, there was a statistical difference between controls and experimentals in concentrations of circulating estrogen; however, these changes were apparently not enough to alter pituitary secretion of follicle-stimulating hormone or luteinizing hormone. These data support the hypothesis that E₂ can induce atresia of the DF in rhesus monkeys by acting locally at the ovary.