Yu-Il Lee

Regulation of NGFI-B expression during the ovulatory process.

NGFI-B is an immediate-early gene that encodes an orphan nuclear receptor. In the rat ovary, the preovulatory surge of LH induces NGFI-B expression in granulosa cells of preovulatory follicles, reaching a peak within 1 h and declining to control levels at 6 h. The LH-stimulated NGFI-B expression is abolished by alpha-amanitin, but superinduced by cycloheximide. Similarly, treatment of human luteinized granulosa cells with LH causes a rapid and transient stimulation of NGFI-B expression. Interestingly, the induction of NGFI-B expression in response to LH stimulation in preovulatory granulosa cells requires signaling through protein kinase Czeta. Furthermore, two other NGFI-B family members, Nurr1 and Nor1, are also rapidly stimulated by LH in granulosa cells of preovulatory follicles through the activation of protein kinase Czeta. The cell-type specific expression and LH induction of NGFI-B suggests a potential role of NGFI-B in the ovulatory process.

Effect of adding non-volatile oil as a core material for the floating microspheres prepared by emulsion solvent diffusion method.

Eudragit® microspheres, to float in the gastrointestinal tract, were prepared to prolong a gastrointestinal transit time. To enhance their buoyancy, non-volatile oil was added to the dispersed phase. When an oil component was not miscible with water, over 90% was entrapped within the microspheres and prolonged the floating time of the microspheres. Depending on the solvent ratio, the morphologies of the microspheres were different and the best result was obtained when the ratio of dichloromethane:ethanol:isopropanol was 5:6:4. As the isopropanol portion increased, the time to form microspheres was delayed and the amount of fibre-like substance produced was decreased, due to the slow diffusion rate of the solvent. Compared with microspheres prepared without non-volatile oil, the release rate of the drug from microspheres was faster in all cases tested, except the microspheres containing mineral oil. The solubility of the drug in the non-volatile oil affected the release profiles of the drugs. The non-volatile oil tends to decrease the glass transition temperature of prepared microspheres and change the release profile. The internal morphology of the microspheres was slightly different depending on the entrapped oil phase used. Tiny spherical objects were present at the inner surface of microspheres and the inside of the shell.Eudragit microspheres, to float in the gastrointestinal tract, were prepared to prolong a gastrointestinal transit time. To enhance their buoyancy, non-volatile oil was added to the dispersed phase. When an oil component was not miscible with water, over 90% was entrapped within the microspheres and prolonged the floating time of the microspheres. Depending on the solvent ratio, the morphologies of the microspheres were different and the best result was obtained when the ratio of dichloromethane:ethanol:isopropanol was 5:6:4. As the isopropanol portion increased, the time to form microspheres was delayed and the amount of fibre-like substance produced was decreased, due to the slow diffusion rate of the solvent. Compared with microspheres prepared without non-volatile oil, the release rate of the drug from microspheres was faster in all cases tested, except the microspheres containing mineral oil. The solubility of the drug in the non-volatile oil affected the release profiles of the drugs. The non-volatile oil tends to decrease the glass transition temperature of prepared microspheres and change the release profile. The internal morphology of the microspheres was slightly different depending on the entrapped oil phase used. Tiny spherical objects were present at the inner surface of microspheres and the inside of the shell.

Expression of peroxiredoxin I regulated by gonadotropins in the rat ovary

Objective Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. Methods Immature rats were injected with 10 IU of pregnant mares serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. Results During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. Conclusion These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.

In Vitro Stimulation of Cumulus-Cell Expansion by Human Cord Serum in Mouse Oocyte–Cumulus Complexes

Purpose:The aim of this study was to test whether human cord serum (HCS) containing gonadotropins has an effect on the expansion ofoocyte—cumulus complexes (OCCs).Methods:The concentration of follicle stimulating hormone (FSH) and luteinizing hormone (LH) was measured in HCS by radioimmunoassay (RIA). After short-term culture (4 hr) with or without OCCs, medium containing 0.4% bovine serum albumin (BSA) as control or 10% HCS was collected and analyzed for its concentration of estradiol, progesterone, and testosterone.Results:The FSH concentration was at the basal level, but the LH level was as high as 142.4 mILU/ml in both natured and denatured serum. Undetectable levels of steroids were observed in control media with or without OCCs. In contrast, a moderate amount of steroid hormones was detected in culture medium containing HCS. OCCs secreted a minute amount of steroid hormones in response to HCS. Similar patterns of cumulus expansion were observable by treatment with HCS, human chorionic gonadotropin (hCG), or HCS plus hCG after 4, 8, or 22 hr of culture. However, no cumulus expansion was observed in controls.Conclusion:These results suggest that LH in HCS induces cumulus expansion but does not affect the secretion of steroid hormones by OCCs during culture.