Kim, Dae-Jung;Kim, Eung-Oh;Park, Min-Woo;Cho, Yong-Chul;Lim, Sang-Gu
Journal of Aquaculture
/
v.19
no.4
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pp.267-274
/
2006
To understand the changes in plasma levels of sex steroids in the wild Japanese eel Anguilla japonica during artificially maturing process, eels received weekly intraperitoneal injections of a water-in-oil (W/O) type emulsion with Freund`s incomplete adjuvant containing salmon pituitary extract (SPE; 20 mg pituitary powder/fish) were examined. In the weekly Eel's Ringer-treated control wild eels, the body weight (BW) changes of fish decreased slowly during the experiment period. Plasma testosterone (T), $estradiol-17{\beta}\;(E_2)$ and $17a,20{\beta}-dihydroxyprogesterone$ (DHP) levels did not change significantly at the end of the experiment. In the weekly SPE-treated silver eels, however, rapid increase in BW changes occurred after 6 to 10 weeks, and the oocytes of all fish were observed to be in the migratory nucleus stage. Furthermore, significant increase in sex steroid hormones (T and $E_2$) levels occurred from 6 weeks. In the weekly SPE-treated yellow eels, the BW changes of fish increased slowly at 6 weeks and then increased. In these fish, the oocytes were at the tertiary yolk globule stage even at the end of the experiment. Plasma sex steroid hormones profiles revealed individual variability in SPE-treated yellow eels. Plasma T and $E_2$ levels significantly increased at 8 weeks and after 6 weeks, respectively, in SPE-treated yellow eels. In the weekly SPE-treated wild eels (silver and yellow eels), however, plasma DHP levels did not change significantly during the experiment period. In silver eel, final maturation could be induced by weekly administration of SPE using W/O type emulsion.
This study was conducted to investigate the hormonal changes in cultured medium during in vitro culture of bovine oviduct epithelial cells (BOEC) supplemented with interleukin (IL)-4 of 0.001, 0.01, 0.1 or 1 ng/ml. BOEC were collected from the oviduct and washed 3 times with 1% antibiotic-mycotic-DMEM medium and cultured at $39^{\circ}C$, 5% $CO_2$, 95% air for 24$\sim$120 hrs. The cultured media were analyzed hormonal changes with hormonal analyzing kit (progesterone (P4), estradiol (E2) : Perkin Elmer, USA) and Transforming growth factor (TGF)-$\beta$ with Eliza kit (Promega, USA). The production of P4 in 0.001 IL-4 was increased as the culture time increased. P4 production was significantly higher in the medium cultured for 120 hrs than 24 hrs (P<0.05). P4 production in 0.01 ng/ml group was similar to that of 0.001 ng/ml. The production of E2 in 0.001 and 0.01 ng/ml groups were increased to 72 hrs like P4 production and showed significantly different between the culture periods (P<0.05). After the culture for 96 hrs, P4 and E2 production were increased to 96 hrs, but decreased at 120 hrs. The production of TGF-$\beta$ showed no changes according to culture period or supplementation of IL-4. In conclusion, the supplementation of IL-4 can increase the production of P4 and E2 and might have important role for the successful pregnancy in bovine.
The aim of this study was to determine the effect of steroids and human chorionic gonadotropin (HCG) on in vitro maturation and ovulation of oocyte in Pseudobagrus fulvidraco. Oocytes were incubated in the media Leibovitz L15 supplemented with the various concentration of $17\alpha,\;20\beta-dihydroxy-4-pregnen-3-one(17\alpha20{\beta}OHP),\;17\alpha-hydroxyprogesterone(17{\alpha}OHP),\;progesterone(P_4),\;estradiol-17\beta(E_2)and\;HCG$. After 60 hours incubation, the maturation ability of oocyte was assessed by the appearance of germinal vesicle breakdown (GVBD). GVBD was significantly enhanced by the addition of $17\alpha20{\beta}OHP,\;17{\alpha}OHP,\;P_4\;and\;HCD(P<0.05)$. The highest CVBD was observed when $17\alpha20{\beta}OHP$ and HCG were supplemented to media. When oocytes were cultured for 16 hours in media containing $10\~1,000\;ng/ml\;17\alpha20{\beta}OHP,\;17{\alpha}OHP\;and\;P_4$, the rate of GVBD in oocytes cultured in the medium supplemented with 100 ng/ml $17\alpha20{\beta}OHP(65\%)$ was significantly higher than that with $17{\alpha}OHP\;(40\%)\;and\;P_4(35\%)$. The efforts of $17\alpha20{\beta}OHP$ and HCG on GVBD were assessed by various concentration of these hormones. When oocytes were cultured for 60 hours in various media containing $1\~1,000\;ng/ml\;17{\alpha}20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG, the GVBD of oocytes was significantly increased in the medium with $10\~100\;ng/ml\;17\alpha20{\beta}OHP$ and 500 IU/ml HCT. When oocytes were cultured in the various media supplemented with $1\~1,000\;ng/ml\;17\alpha20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG for 60 hours, the media with $1\~100\;ng/ml\;17\alpha20{\beta}OHP\;or\;50\~1,000IU/ml$ HCG significantly increased in the rate of ovulation. However supplementation with $1,000\;ng/ml\;17\alpha20{\beta}OHP$or 5 IU/ml HCG did not improve the rate of ovulation compared to controls. This results indicate that supplementation of steroid and HCG except $E_2$ can improve the in vitro maturation and ovulation of oocyte in P. fulvidrac; HCG and $17\alpha20{\beta}OHP$ may be more effective than other steroids on oocyte maturation and ovulation in P. fulvidraco.
Journal of Fisheries and Marine Sciences Education
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v.26
no.5
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pp.935-940
/
2014
To investigate effect of on sexual maturation in spotted seabass, Lateolabrax maculatus the sexual maturation, photoperiods were established to natural photoperiod, 9L:15D, 11L:13D, 13L:11D and 15L:9D. After rearing for 90 days from August 11 to November 10, gonadosomatic index (GSI) and hormones were analyzed. In the female, GSI was $1.07{\pm}0.09%$ in the natural photoperiod, $1.93{\pm}0.06%$ in 9L:15D, while the GSI of male was $0.75{\pm}0.05%$ in the natural photoperiod, $1.89{\pm}0.12%$ in 9L:15D and $0.22{\pm}0.05%$ in 15L:9D and the lowest in 15L:9D, respectively. The blood estradiol (E2) of female was $226.0{\pm}13.6pg/ml$ in the natural photoperiod, and the highest value of $296.3{\pm}15.0pg/ml$ in 9L:15D. The male $17{\alpha}20{\beta}OHP$ was similar to the female with $196.3{\pm}17.0pg/ml$ in the natural photo period and $133.0{\pm}15.5pg/ml$ in 9L:15D. The testosterone (T) of male was $259.3{\pm}33.1pg/ml$ in the natural photoperiod, but the hightest value of $370.5{\pm}27.5pg/ml$ in 9L:15D. These results provide basic information from sexual maturation of spotted seabass.
We report on two cases of pericentric inversion of X chromosome. The cases were found in a 40-year-old man with azoospermia and in a family of a 38-year-old pregnant woman. The first case with 46,Y,inv(X)(p22.1q27) had concentrations of LH, prolactin, estradiol, and testosterone that were within normal ranges; however, FSH levels were elevated. Testis biopsy revealed maturation arrest at the primary and secondary spermatocytes without spermatozoa. There were no microdeletions in the 6 loci of chromosome Y. For the second case, the cytogenetic study of thepregnant woman referring for advanced maternal age and a family history of inversion X chromosome was 46,X,inv(X)(p22.11q27.2). The karyotype of her fetus was 46,X,inv(X)(p22.1q27). Among other family members, the karyotypes of an older sister in pregnancy and her fetus were 46,X,inv(X)(p22.11q27.2), and 46,Y,?inv(X), respectively. The proband's father was 46,Y,inv(X)(p22.11q27.2). All carriers in the family discussed above were fertile and phenotypically normal. In addition, the ratio of inactivation of inv(X) by RBG-banding was discordant between the two sisters, with the older sister having only 4.1% of cells carrying inactivated inv(X) while the proband had a 69.5% incidence of late replicating inv(X). Therefore, we suggest that the cause of azoospermia in the first case might be related to inversion X chromosome with positional effect. Also, the family of the second case showing normal phenotype of the balanced inv(X) might be not affected any positional effect of genes.
Concerns about endocrine disrupting chemicals emitted from humans and animals have been increased because these compounds are detected at very low levels in environment and adversely affect on indigenous fauna. To date, there is little information regarding the concentration of these compounds in animal wastes. In this study, the female hormones, $17\beta-estradiol$ (E2), estrone (E1) and estriol, were measured to provide baseline data in animal wastes. Samples were collected from animal waste storage, methane digester and sludge separated wastewater and analyzed by gas chromatography-mass spectrometry. To measure the mass ratios of estrogen to macronutrients, nitrogen and phosphorous were also determined. Sample collected from animal waste storage had the highest estrogen concentration (98.7 ${\mu}g/L$), while sludge separated wastewater had the lowest concentration (3.4 ${\mu}g/L$). The mean concentrations of E2 and E1 in waste storage sample were (6.8 ${\mu}g/L$) and (68.7 ${\mu}g/L$), respectively. In sludge separated wastewater, the mean concentration of both E2 and E1 were reduced to (2.6 ${\mu}g/L$) and (1.9 ${\mu}g/L$), respectively. However, estriol was not detected in any of the samples collected. Mean ratios of E2 and E1 to macronutrients were significantly different between the methane wastewater and sludge separated wastewater owing to elimination of solid particles.
Sex steroids are known to be involved in skeletal muscle development (anabolic effect) and are frequently used in medicines. It has been known that pork contains a variety of steroids that are mainly synthesized in the gonads (testis and ovary). Thus, the present study was conducted to evaluate the effects of anabolic steroids of pork on the proliferation and differentiation of myogenic satellite cells (MSC). Three different methods (M1, M2, and M3) were developed for the isolation and purification of steroids from porcine tissues. Among three extraction methods that we developed, M3 was the best method with respect to the quantities of steroids and the induction of MSC proliferation. Hormonal analysis showed that the steroid hormone levels were the highest in muscle and fat of intact male than those of castrated males and females. In addition, the highest serum levels of nandrolone and testosterone were detected in intact males, whereas estrone and $17{\beta}$-estradiol levels were similar in the entire experimental serum samples. Expression of androgen receptor (AR), myoD, desmin, and myogenin in bovine muscle cells were significantly up-regulated by the treatment of steroid extracts. The highest increas of myogenin and AR mRNA abundance were observed in the MSCs treated with M3 extract (p<0.001). Altogether, the present research showed the positive effect of steroids on MSC proliferation and differentiation in vitro. These results would certainly imply a beneficial effect of pork consumption on human muscle development.
GnRH and its receptor are known to express locally in the ovary and to regulate the ovarian function by affecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor is not yet known. In this study, we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ after treatment with 5, 50, and 100 ng/$m\ell$ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/$m\ell$ GnRH. In addition, we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by GnRH treatment, whereas $E_2$ production was not changed. We also demonstrated that FSH inhibits the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of the ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.
Cortisol is present in high concentration in the ovary and its receptor is expressed in the ovarian cells. Moreover, cortisol is known to have a role in steroid synthesis and cell metabolism in human granulosa and lutein cells. However, little is known of the role of cortisol presenting in high concentration in the follicles after LH surge on the granulosa-lutein cells. Therefore, the this study we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ in the granulosa-lutein cells that are obtained during oocyte-retrieval after treatment with 5, 50, and $500{\mu}g/m\ell$ cortisol and 1 IU/$m\ell$ FSH. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in 50 and $500{\mu}g/m\ell$ cortisol treated cells. We found, however, that FSH did not suppress the apoptosis of the cells induced by cortisol. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by cortisol treatment, whereas $E_2$ was not changed. We also demonstrated that FSH did not inhibit the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present study suggests that cortisol of high concentration could cause the apoptosis of human granulosa-lutein cells by suppressing the production of $P_4$. However, we need more studies to elucidate the mechanism by which cortisol induces apoptosis in human granulosa-lutein cells in view of the fact that our results are inconsistent with previous reported data.
This study was performed to investigate the correlations between steroids and TGF-${\beta}_1$ levels and delayed parturition in SCNT clone calving. The recipients pregnant by AI were used as control (AI-R). All AI-R were labored by natural delivery (n=5, day $284{\pm}0.71$ of pregnancy). The recipients pregnant by SCNT embryo (SCNT-R) showing no signs of delivery about 10 days after expected date were operated by Caesarean section (n=5, day 292). The blood and placentome samples were obtained and weighed at parturition. The concentrations of plasma progesterone (P4) and Estradiol-$17{\beta}$ (E2) were measured by radioimmunoassay (RIA). The levels of plasma and placental TGF-${\beta}_1$ levels were examined by ELISA. The placentomes from SCNT-R were overweight (p<0.05) compared to those of AI-R. The plasma P4 (p<0.01) level in SCNT-R at parturition was significantly higher compared to that of AI-R. In contrast, the plasma E2 level in the SCNT-R was significantly lower compared to that of AI-R (p<0.05). The plasma and placental TGF-${\beta}_1$ protein levels in the SCNT-R were significantly higher than those of AI-R at parturition, respectively (p<0.01). Based on these results, aberrant expressions of steroid hormones and high levels of plasma and placental TGF-${\beta}_1$ protein at parturition may be one of the key indicators on delayed parturition of SCNT clone calving.
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