The control mechanism of gonadotropin-releasing hormone (GnRH) on gonadotropin (GTH) release was studied using cultured pituitary cell or cultured whole pituitary obtained from Testosterone (T) treated and control immature rainbow trout. The release of FSH was not changed by salmon type GnRH (sGnRH), chiken-II type (cGnRH-II), GnRH analogue ([des-$Gly^{10}D-Ala^6$] GnRH ethylamide) and GnRH antagonist ([Ac-3, 4-dehydro-$Pro^1$, D-p-F-$Phe^2$, D-$Trp^{3,6}$] GnRH) in cultured pituitary cells of T-treated and control fish. Indeed, FSH release was not also altered by sGnRH in cultured whole pituitary. All tested drugs had no effect on the release of LH in both culture systems of control fish. The levels of LH, in contrast, such as the pituitary content, basal release and responsiveness to GnRH were increased by T administration in both culture systems. In addition, the release of LH in response to sGnRH or cGnRH-II induced in a dose-dependent manner from cultured pituitary cells of T-treated fish, but which is not significantly different between in both GnRH at the concentration examined. Indeed, LH release was also increased by sGnRH in cultured whole pituitary of T-treated fish. GnRH antagonist suppressed the release of LH by sGnRH ($10^{-8}\;M$) and GnRH analogue ($10^{-8}\;M$) stimulation in a dose-dependent manner from cultured pituitary cells of T-treated fish, and which were totally inhibited by $10^{-7}\;M$ GnRH antagonist. These results indicate that the sensitivity of pituitary cells to GnRH is elevated probably through the T treatment, and that GnRH is involved in the regulation of LH release. GnRH-stimulated LH release is inhibited by GnRH antagonist in a dose-dependent manner. The effects of gonadal steroids on FSH levels are less clear.
Objective: To evaluate the ability of serum anti-M$\ddot{u}$llerian hormone (AMH), FSH, and age to clinically predict ovarian response to controlled ovarian hyperstimulation (COH) in IVF patients with endometriosis. Methods: We evaluated 91 COH cycles, including 43 cycles with endometriosis (group I) and 48 cycles with male factor infertility (group II) from January to December, 2010. Patients were classified into study groups based on their surgical history of endometriosis-group Ia (without surgical history, n=16), group Ib (with a surgical history, n=27). Results: The mean age was not significantly different between group I and group II. However, AMH and FSH were significantly different between group I and group II ($1.9{\pm}1.9$ ng/mL vs. $4.1{\pm}2.9$ ng/mL, $p$ <0.01; $13.1{\pm}7.2$ mIU/mL vs. $8.6{\pm}3.3$ mIU/mL, $p$ <0.01). Furthermore, the number of retrieved oocytes and the number of matured oocytes were significantly lower in group I than in group II. In group II, AMH and FSH as well as age were significant predictors of retrieved oocytes on univariate analysis. Only the serum AMH level was a significant predictor of poor ovarian response in women with endometriosis. Conclusion: Serum AMH may be a better predictor of the ovarian response of COH in patients with endometriosis than basal FSH or age. AMH level can be considered a useful clinical predictor of poor ovarian response in endometriosis patients.
Objectives: Polycystic ovary syndrome (PCOS) has the feature of excessive LH, hyperandrogenism and disturbance of folliculogenesis. Also, insulin, IGF-I and IGFBP-l are involved in the pathogenesis of PCOS. Various surgical and medical therapies have been used and the action mechanisms are related to the endocrine effect. Laparoscopic ovarian electrocautery or laser vaporization is effective in the restoration of ovulation and normal menstrual cycle with minimal invasive procedure especially in the patients resistant to medical therapy. Clomiphen citrate (CC) is used for the ovulation induction in pcas and the resistance is known to be related to insulin, IGF-I, IGFBP-l levels. This study was performed to evaluate the effect of the laparoscopic laser vaporization on the levels of LH, FSH, testosterone, IGF-I and IGFBP-l and on the ovarian response to clomiphen citrate in patients with CC-resistant PCOS. Materials and Methods: The fasting basal serum LH, FSH, testosterone, IGF-I and IGFBP-l level were measured in 10 PCOS patients with CC-resistance and 7 normal controls with regular menstrual cycle. In PCOS, after laparoscopic $CO_2$ laser vaporization, endocrine levels were measured in 1 week interval for 4 weeks and then compared with preoperative levels. Results: In PCOS group, mean serum LH/FSH ratio, testosterone, IGF-I levels were higher and IGFBP-l level was lower than control. LH/FSH ratio decreased from $2.51{\pm}0.67$ to $1.7{\pm}0.6$ (p<0.05) in 2 weeks, to $0.56{\pm}0.2$ (p<0.01) in 3 weeks and to $1.41{\pm}0.3$ (p<0.01) in 4 weeks after operation. Testosterone level decreased from $1.51{\pm}0.82ng/ml$ to $0.65{\pm}0.34ng/ml$ (p<0.05) in 2 weeks, to $0.56{\pm}0.67ng/ml $(p<0.01) in 3 weeks after operation. IGF-I level also decreased from $436{\pm}47.5{\mu}g/l$ to $187{\pm}38{\mu}g/l$ (p<0.0l) in 1 week, to $167{\pm}42{\mu}g/l$ (p<0.01) in 2 weeks, $179{\pm}55{\mu}g/l$ (p<0.01) in 3 weeks and to $120{\pm}43{\mu}g/l$ (p<0.01) in 4 weeks after operation. IGFBP-l level showed no significant change. In 8 of 10 PCOS patients, ovulation was induced with low dose clomiphen citrate. Conclusion: Laparoscopic $CO_2$ laser vaporization restores normal menstrual cycle and ovulation through endocrine effect of decreasing LH/FSH ratio, testosterone and IGF-I level and increases the response to CC. Therefore it is useful for restoration of normal menstruation and induction of ovulation in CC resistant PCOS patients.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
Lee, Woo Seok;Sang, Jae Hong;Kim, Jae Joon;Kim, Gwang June;Kim, Dong Ho;Lee, Sang Hun
Clinical and Experimental Reproductive Medicine
/
v.33
no.3
/
pp.149-157
/
2006
Objective: This study was to investigate the clinical efficiency of clomiphene citrate/GnRH antagonist protocol comparing with the clomiphene citrate only protocol in infertile women with normal ovulatory cycles. Method: Among 116 patients, 43 were received assisted reproductive technologies using natural ovulatory cycle, 38 and 35 were received clomiphene citrate only protocol and clomiphene citrate/GnRH antagonist combined protocol, respectively, and the clinical results were compared and analyzed Results: In each group, basal levels of LH, FSH, $E_2$ and FSH, $E_2$ on hCG day injected were not different, but LH level and endometrial thickness on hCG injected day were decreased significantly and the pregnancy rate was increased significantly in clomiphene citrate/GnRH antagonist group. Conclusion: The pregnancy rate was increased significantly in clomiphene citrate/GnRH antagonist group compared with natural ovulatory cycle and clomiphene citrate only group.
Objective: To determine the cutoff value of clomiphene citrate challenge test(CCCT) that can predict the normal and abnormal(diminished) ovarian response group and to assess the usefulness of CCCT as a predictor of ovarian reserve. Materials and Methods: From March 1994 to Februry 1996, CCCT was performed to 129 infertile patients and among them, 20 patients whose basal FSH on the third day of menstrual cycle was more than 20 mIU/ml were excluded. At the same time, the same CCCT was performed to the fifteen healthy volunteers with proven fertility to determine the cutoff value of CCCT. Results; 1) A FSH value higher than 23.4 mIU/ml, measured on the 10th day of menstrual cycle, was defined as a abnormal ovarian response. The cutoff value of 23.4 mIU/ml is more than 2 standard deviations(SD) above the mean value of 15 healthy women after CCCT. 2) The abnormal CCCT group, the subpopulation with a FSH value of 23.4 mIU/ml or more, was 7.3%(8/109) and their mean age was higher than the normal CCCT group($36.5{\pm}4.5$ vs. $32.9{\pm}4.8$, P = 0.059). And the percentage of the patients older than 35 years of the abnormal CCCT group was significantly higher than that of the normal CCCT group(62.5% vs. 38.6%, p <0.05). 3) There was no correlation between the hormone values of the third day and the 10th day of menstrual cycle before and after CCCT except between FSH of the third day and the 10th day. Conclusion: The CCCT is a good method to predict the individual ovarian response to COH for ART, especially in the patients who has no other abnormal findings that predict poor prognosis. And it is neccessary to determine the cutoff value of CCCT by the large numbers of randomized study, and combining the previously proven prognostic factors, it can be applicated in many individual centers for evaluate the ovarian response before ART program.
The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.
Recently, As the prevalence of precocious is increased in Korea This study was evaluated the physical growth and female sex hormone characteristics of girls with Idiopathic precocious puberty. Retrospective analysis about the medical record of 113 girls for evaluation of signs of precocious puberty except for organic causes was done. Physical growth features and Female sexual hormone were analyzed. Height SDS and Weight SDS were significantly increased in true sexual precocity compared to pseudo sexual precocity. Hormone studies showed that the level of basal LH, FSH, E2 was significantly higher in true precocious puberty. The finding of this study suggest that we need to develop intervention about physical and psychological problems for precocious puberty girls.
To investigate the factors that affect the fertilization and cleavage rates of mature oocytes, 44 patients undergoing controlled ovarian hyperstimulation(COH) with FSH/hMG/hCG regimen for IVF - ET were analyzed. During follicular phase, serum LH levels were measured by radioimmunoassay and bioassay. Based on the mean follicular immunoactive LH(i-LH) and bioactive LH(b-LH) levels, patients were divided into 3 groups, respectively. There were no significant differences in basal serum FSH levels on menstrual cycle day 3, serum estradiol($E_2$) and progesterone ($P_4$) levels on the day of hCG administration, and the numbers of follicles aspirated and oocytes retrieved among groups. In relation to the mean follicular i-LH levels, the fertilization and cleavage rates of mature oocytes did not show a significant difference among groups. However, in groups with higher mean follicular b-LH levels, the fertilization and cleavage rates were reduced significantly. During late follicular phase, day-to-day variance in b-LH levels was not significant, but there was a significant difference among groups. There was no significant correlation between serum P. and b-LH levels. These data suggest that the fertilization and cleavage rates of mature oocytes are adversely affected by the raised mean follicular b-LH levels, and monitoring of serum b-LH levels is more useful in COH when compared with i-LH. It appears that the reduced rates are not due to the attenuated endogenous LH surge.
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