Origin of ovarian follicle cells in birds

Summary In the embryonic Japanese quail ovary, transplanted on chicken chorioallantoic membrane (CAM), follicle cells are derived from somatic cells of the ovarian surface epithelium. No evidence was found for a contribution of other cell groups of the quail ovary in the formation of follicle cells. This may be demonstrated on PAS stained sections, by following the transfer of carbon particles, initially applied on the surface epithelium.The author is very grateful to Prof.L. Vakaet, R.U.C.A. Antwerp, for his valuables suggestions, and to MissC. van Hoecke for her technical assistance.     

Penetration of melanocytes from embryonic Japanese quail peritoneum into associated embryonic avian gonads,grown on chicken chorioallantoic membrane

Summary After association on chorioallantoic membrane (CAM) of an embryonic bird testis with pigmented peritoneum from a Japanese quail embryo, numerous melanocytes penetrate in its interstitial tissue. If, instead of a testis, an ovary is transplanted under similar conditions, then the melanocytes may be found in the medulla or between the secondary sex cords at the rim of the ovary.The author is very grateful to Prof.L. Vakaet, R. U. C. A. Antwerp, for his valuable suggestions, and to MissC. Vaniioecke for her technical assistance.     

Development and use of shell-less quail chorio-allantoic-membrane cultures to study developing skeletal tissues; a qualitative study

Summary A technique is described for in vitro culture of the quail embryo from the 1st to the 18th day of development. The embryos are cultured in Teflon hammocks, suspended in glass supports and kept in a humidified atmosphere at 36.5°C. The quail CAM is used as support and cell source for developing non-quail cartilage and bone. The quail cells can be identified histologically and easily recognized by Feulgen-staining which is demonstrated in the presence of quail chondro- or osteoclasts in a mouse long bone rudiment cultured on the CAM.     

Development and use of shell-less quail chorio-allantoic-membrane cultures to study developing skeletal tissues; a qualitative study

A technique is described for in vitro culture of the quail embryo from the 1st to the 18th day of development. The embryos are cultured in Teflon hammocks, suspended in glass supports and kept in a humidified atmosphere at 36.5 degrees C. The quail CAM is used as support and cell source for developing non-quail cartilage and bone. The quail cells can be identified histologically and easily recognized by Feulgen-staining which is demonstrated in the presence of quail chondro- or osteoclasts in a mouse long bone rudiment cultured on the CAM.     

Stérilisation de l'ovaire gauche du Poulet et de la Caille par l'action de la thymidine-3H pendant l'ovogenèse

Summary After successive in ovo³H-thymidine pulses during the period of oogenesis complete sterilization of the left ovary of the female chicken and quail have been obtained at the moment of hatching.     

3H-thymidine incorporation (DNA synthesis) and radiotoxicity in the ovary of the Japanese quail before and during follicle formation

No evidence was found for ribosomal DNA amplification in the oocytes of the Japanese quail, before or during folliculogenesis. DNA synthesis in the somatic cells, involved in follicle formation, starts at the medullar side of the basement membrane. The localized sterilization of the quail ovary after administration of 3H-thymidine (3H-TdR) seems to be due to radiation-induced lesions in the follicle forming somatic cells, rather than to direct radiation damage of the oocyte.     

3H-Thymidine incorporation (DNA synthesis) and radiotoxicity in the ovary of the Japanese quail before and during follicle formation

Summary No evidence was found for ribosomal DNA amplification in the oocytes of the Japanese quail, before or during folliculogenesis. DNA synthesis in the somatic cells, involved in follicle formation, starts at the medullar side of the basement membrane. The localized sterilization of the quail ovary after administration of³H-thymidine (³H-TdR) seems to be due to radiation-induced lesions in the follicle forming somatic cells, rather than to direct radiation damage of the oocyte.The author is very grateful to Prof. Dr L. Vakaet, R.U.C.A.—Antwerpen, for his valuable suggestions and to Mrs D. De Wolf-Van Rompaey for her excellent technical assistance.     

Antiproliferative effect of β-elemene in chemoresistant ovarian carcinoma cells is mediated through arrest of the cell cycle at the G2-M phase

Elemene is a natural antitumor plant drug. However, the effect of elemene on cell growth in ovarian cancer is unknown. In this study, we show that -elemene inhibited the proliferation of cisplatin-resistant human ovarian cancer cells and their parental cells, but had only a marginal effect in human ovary cells, indicating differential inhibitory effects on cell growth between ovarian cancer cells and normal ovary cells. We also demonstrated for the first time that -elemene markedly enhanced cisplatin-induced growth inhibition in resistant cells compared to sensitive cells. In addition, cell cycle analysis revealed a synergistic effect of -elemene and cisplatin on the induction of cell cycle G2-M arrest in our resistant ovarian carcinoma cells. Furthermore, we showed that treatment of these cells with both drugs downregulated cyclin B1 and Cdc2 expression, but elevated the levels of p53, p21waf1/cip1, p27kip1 and Gadd45. Finally, the combination of -elemene and cisplatin was found to increase the phosphorylation of Cdc2 and Cdc25C, which leads to a reduction in Cdc2-cyclin B1 activity. These novel findings suggest that -elemene sensitizes chemoresistant ovarian carcinoma cells to cisplatin-induced growth suppression partly through modulating the cell cycle G2 checkpoint and inducing cell cycle G2-M arrest, which lead to blockade of cell cycle progression.Received 19 January 2005; accepted 5 February 2005     

Production of transgenic birds

The avian embryo presents a tremendous challenge for those interested in accessing and manipulating the avian germ line. By far the most successful method of gene transfer is by retrovirus vector. The efficacy of retrovirus vectors has been demonstrated by germ line insertion of replication-competent retroviruses as well as the insertion of replication-defective retrovirus vectors carrying bacterial marker genes. Retroviral vectors have also been shown to be useful for the transfer and expression of genes in somatic cells. Further, germ line transgenesis has been reported in both the chicken and the Japanese quail. In addition, several alternative gene transfer methods are under development. These include transfection of avian sperm, development of germ line chimeras using primordial germ cells and blastodermal cells, and the development of embryonic stem cell lines. Potentially, basic research and the poultry industry will derive substantial benefit from this revolutionary technology.     

Production of transgenic birds

The avian embryo presents a tremendous challenge for those interested in accessing and manipulating the avian germ line. By far the most successful method of gene transfer is by retrovirus vector. The efficacy of retrovirus vectors has been demonstrated by germ line insertion of replication-competent retroviruses as well as the insertion of replication-defective retrovirus vectors carrying bacterial marker genes. Retroviral vectors have also been shown to be useful for the transfer and expression of genes in somatic cells. Further, germ line transgenesis has been reported in both the chicken and the Japanese quail. In addition, several alternative gene transfer methods are under development. These include transfection of avian sperm, development of germ line chimeras using primordial germ cells and blastodermal cells, and the development of embryonic stem cell lines. Potentially, basic research and the poultry industry will derive substantial benefit from this revolutionary technology.     

La synthèse du DNA dans les cellules germinales de l'ovaire de poulet pendant la dernière semaine de la vie embryonnaire

Summary DNA synthesis in the germ cells of the ovary of the chicken embryo during the last week of incubation is studied in vivo and in vitro by tritium autoradiography.     

An improved method of transplantation on chicken chorioallantoic membrane (CAM)

Summary An improved method for grafting on the chicken CAM is described and compared with other CAM grafting methods.The author is very grateful to Prof. L. Vakaet, RUCA Antwerpen, to Dr and Mrs M. Mareel, RUG Gent, for their valuable suggestions and to Mrs S. De Wolf-Van Rompaey for her excellent technical assistance. The author is also indebted to Mr F. De Bruyn for making the drawing.     

Transplantations interspécifiques de gonades embryonnaires de Phasianidés

Summary When indifferent gonads from red-legged partridges, Japanese quail or pintado embryos were transplanted into the coelomic cavity of early chick embryos, they differentiated into ovaries and testes, regardless of the sex of the host. Testicular transplants brought about the retrogression of both Müllerian ducts in female hosts, and ovarian transplants the feminization of the left testis in male hosts. This demonstrates hormonal activity of the embryonic gonads in the 3 species under study. Since the embryonic sex hormones show inter-specific activity, they may be of identical chemical nature.

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Avec la collaboration technique deA. Zeis.     

Differentiating abilities of avian somatopleural mesoderm

Summary Quail-to-chick grafting experiments were performed on 2-day embryos in order to test the differentiating abilities of the somatopleure. After orthotopic and heterotopic transplantations of different parts of quail somatopleural mesoderm into chick embryos it is demonstrated that avian somatopleural cells differentiate into skeletal elements, smooth muscles, tendons and connective tissues. However, skeletal muscle fibres do not originate from somatopleural cells.Supported by a grant from the Deutsche Forschungsgemeinschaft (Ch 44/3).     

Gene conversion in the chicken immunoglobulin locus: A paradigm of homologous recombination in higher eukaryotes

Gene conversion was first defined in yeast as a type of homologous recombination in which the donor sequence does not change. In chicken B cells, gene conversion builds the antigen receptor repertoire by introducing sequence diversity into the immunoglobulin genes. Immunoglobulin gene conversion continues at high frequency in an avian leukosis virus induced chicken B cell line. This cell line can be modified by homologous integration of transfected DNA constructs offering a model system for studying gene conversion in higher eukaryotes. In search for genes which might participate in chicken immunoglobulin gene conversion, we have identified chicken counterparts of the yeastRAD51, RAD52, andRAD54 genes. Disruption and overexpression of these genes in the chicken B cell line may clarify their role in gene conversion and gene targeting.     

Mechanism of neurogenesis in adult avian brain

Summary Adult neurogenesis in birds offers unique opportunities to study basic questions addressing the birth, migration and differentiation of neurons. Neurons in adult canaries originate from discrete proliferative regions on the walls of the lateral ventricles. They migrate away from their site of birth, initially at high rates, along the processes of radial cells. The rates of dispersal diminish as the young neurons invade regions devoid of radial fibers, probably under the guidance of other cues. The discrete sites of birth in the ventricular zone generate neurons that end up differentiating throughout the telencephelon. New neurons may become interneurons or projection neurons; the latter connect two song control nuclei between neostriatum and archistriatum. Radial cells, that in mammals disappear as neurogenesis comes to an end, persist in the adult avian brain. The presence of radial cells may be key to adult neurogenesis. Not only do they serve as guides for initial dispersal, they also divide and may be the progenitors of new neurons.     

Mechanism of neurogenesis in adult avian brain

Adult neurogenesis in birds offers unique opportunities to study basic questions addressing the birth, migration and differentiation of neurons. Neurons in adult canaries originate from discrete proliferative regions on the walls of the lateral ventricles. They migrate away from their site of birth, initially at high rates, along the processes of radical cells. The rates of dispersal diminish as the young neurons invade regions devoid of radial fibers, probably under the guidance of other cues. The discrete sites of birth in the ventricular zone generate neurons that end up differentiating throughout the telencephalon. New neurons may become interneurons or projection neurons; the latter connect two song control nuclei between neostriatum and archistriatum. Radial cells, that in mammals disappear as neurogenesis comes to an end, persist in the adult avian brain. The presence of radial cells may be key to adult neurogenesis. Not only do they serve as guides for initial dispersal, they also divide and may be the progenitors of new neurons.     

Ovarian LDH activity in gonadotropin-treated immature rats.

Lactate dehydrogenase (LDH) activity was studied in the ovaries of immature rats treated with pregnant mare serum gonadotropin (PMS). LDH activity increased sharply at 36 h after PMS injection in the ovarian tissue as well as in the blood. It was suggested that the increase of LDH activity in the ovary may be related to its increasing ability to secrete estrogen.     

Identification of cancer stem cell-like cells from human epithelial ovarian carcinoma cell line

Cancer stem cells (CSCs) play an important role in the development, invasion, and drug resistance of carcinoma, but the exact phenotype and characteristics of ovarian CSCs are still disputable. In this study, we identified cancer stem cell-like cells (CSC-LCs) and investigated their characteristics from the ovarian adenocarcinoma cell line 3AO. Our results showed that CSC-LCs were enriched in sphere-forming test and highly expressed CD44⁺CD24⁻. The spheres and CD24⁻ cells possessed strong tumorigenic ability by transplantation into nonobese diabetic/severe combined immunodeficient mice. CD44⁺CD24⁻ cells expressed stem cell markers and differentiated to CD44⁺CD24⁺ cells by immunofluorescence assay and fluorescence-activated cell-sorting analysis. In vitro experiments verified that CD44⁺CD24⁻ cells were markedly resistant to carboplatin and paclitaxol. In conclusion, our study identifies the CD44⁺CD24⁻ phenotype, self-renewal, high tumorigenicity, differentiation potential, and drug resistance of ovarian CSC-LCs. Our findings may provide the evidence needed to explore a new strategy in the treatment of ovarian cancer.     

Cellular turnover and extracellular matrix remodeling in female reproductive tissues: functions of metalloproteinases and their inhibitors

Female reproductive tissues possess a unique ability to accommodate a remarkable amount of cell turnover and extracellular matrix (ECM) remodeling following puberty. Cellular structures within ovary, uterus, and mammary tissue not only change cyclically in response to ovarian hormones but also undergo differentiation during pregnancy, and eventually revert to that resembling the pre-pregnant stage. Cell proliferation, apoptosis, invasion, and differentiation are integral cellular processes that are precisely regulated in reproductive tissues, but become dysregulated in pathologies such as cancer. Explicit reorganization of ECM and basement membranes is also critical to preserve the form and function of these tissues. Here we review the evidence that coordinated spatiotemporal expression patterns of matrix metalloproteinase (MMP) genes and their tissue inhibitors (TIMPs) are important in cell and ECM turnover of the ovary, uterus, and mammary tissues. We discuss how perturbation in these gene families may impact the biology of these reproductive tissues and the factors implicated in the control of MMP and TIMP gene expression. The observed trends in MMP and TIMP expression involved in ovarian and mammary carcinomas are also presented.