The dependence of activities of Aspergillus phoenicis on the culture conditions in the progesterone transformation reaction was investigated. In the beginning of the reaction, $6{\beta},\;11{\alpha}-dihydroxyprogesterone$ was not produced even at high concentration of $11{\alpha}-hydroxyprogesterone$. However, large amount of the product was obtained after the complete exhaustion of progesterone. When spores of A.phoenicis replaced mycelia as enzyme source, $11{\alpha}-hydroxyprogesterone$ was produced after a considerably long indyction period, and its maximum production rate followed the exponential growth phase. The $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone$ continued, even after the stationary growth phase. A. phoenicis showed high enzyme activity for these reactions when the phosphate buffer solutions were used in place of the ordinary culture medium. The buffer solutions of low pH gave more yield of $11{\alpha}-hydroxyprogesterone$ than those of high pH. However, the addition of flucose to the buffer solutions did not activate the transformation reaction. The presence of progesterone seems to be necessary for the induction of enzymes for the $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone\;since\;6{\beta},\;11{\alpha}-dihydroxyprogesterone$ is not produced in the reaction medium containing only $11{\alpha}-hydroxyprogesterone$ as a substrate.
It is generally accepted that chronic stress impairs female reproduction. It negatively affects ovarian function and the number of ovulated oocytes. Chronic stress lowers the number of retrieved oocytes. Ovarian follicular development is regulated by both pituitary-derived gonadotropins and intraovarian regulatory factors. The main corticosteroids are cortisol, cortisone, 11-deoxycortisol and corticosterone, cortisol being one of the most commonly used welfare and stress physiological indicator. In this study, we investigated the effect of cortisol level on progesterone patterns and ovulation in the dog. Cortisol and progesterone level of serum were analyzed by radioimmunoassay. The day of ovulation was considered as the day when serum progesterone concentration was 6.0~8.0 ng/ml. In vivo dog oocytes were collected by flushing oviducts of mixed-breed bitches at three days after ovulation. We classified dogs as having group 1 (cortisol level, 0 ${\leq}$ or < $2{\mu}g/dl$), group 2 (corisol level, 2 ${\leq}$ or < $4{\mu}g/dl$), group 3 (cortisol level, 4 ${\leq}$ or < $6{\mu}g/dl$) and group 4 (cortisol level, $6{\mu}g/dl$${\leq}$). The patterns of progesterone were not different in four cortisol groups. The average numbers of retrieved oocytes was not different in four cortisol groups. These results suggest that different cortisol levels on estrus dogs do not affect ovulation, number of ovulated oocytes and progesterone changes.
This experiment was conducted to examine the $PGF{2{\alpha}}$-induced changes in concentrations of ovarian and pituitary hormones of Korean native goats. Each goats received two injections of $PGF{2{\alpha}}$ (5mg each ; 3 hours apart) on day 10 of the estrous cycle. Jugular venous blood samples were collected at 0, 3, 6, 12, 24, 48 and 72 hours postinjection for quantification of LH, FSH, prolactin, progesterone and estradiol-$17{\beta}$. The results were summarized as follows ; The blood serum concentration of progesterone was decreased from pretreatment level of $4.15{\pm}1.8ng/ml$ to $2.52{\pm}1.2ng/ml$ (about 60%) within 3 hours and to $0.81{\pm}0.3ng/ml$ at 12 hours of the $PGF{2{\alpha}}$ injection. After 12 hours, the concentrations of progesterone were less than 1.02ng/ml by 72 hours postinjection. The concentrations of estradiol-$17{\beta}$ following treatment increased (p < 0.05) over the 72 hours. Initial concentration of LH was $3.0{\pm}0.3{\mu}IU/ml$. After treatment with $PGF{2{\alpha}}$, concentrations of LH increased within 12 hours but declined 12 and 72 hours from $4.1{\mu}IU/ml$ to $2.5{\mu}IU/ml$. Prior to administration of $PGF{2{\alpha}}$, mean concentration of FSH was $3.5{\pm}0.5{\mu}IU/ml$. Concentrations of FSH declined over time in goats treated with $PGF{2{\alpha}}$ on day 10 postestrus. The mean prolactin concentrations in the blood serum after $PGF{2{\alpha}}$ treatment were not significantly different from those of the pretreatment. It is concluded that the initial increase in LH is dependent on a decrease in serum progesterone and differences in patterns of secretion of gonadotropins might be caused by differences in progesterone or progesterone-estradiol ratio when luteal regression is induced on day 10 of the estrous cycle.
Among fatty acid families, the polyunsaturated fatty acids were demonstrated to be mediators in various reproductive processes as precursor of steroid hormone (via cholesterol) and prostaglandins (via arachidonic acid), and in the last decade, major research was focused on the effects of omega-6 and especially omega-3 fatty acid. Eicosapentaenoic acid, the longest members of omega-3 fatty acid family, can be produced by a series of desaturation and elongation reactions from shorter member such as α-Linolenic acid. However, very few studies have provided detailed descriptions of Eicosapentaenoic acid effects and mechanisms of action in mammalian oocytes. The purpose of this study was to evaluate the effect of Eicosapentaenoic acid supplementation on in vitro maturation and developmental potential of porcine oocytes. Various concentrations of Eicosapentaenoic acid was added into in vitro maturation medium, and we evaluated the degree of cumulus expansion, nuclear maturation rate, blastocysts quality, and levels of prostaglandin E2, 17β-estradiol, progesterone in the spent medium. High doses (100 μM) of Eicosapentaenoic acid supplementation significantly inhibited cumulus expansion and oocyte nuclear maturation, and prostaglandin E2 synthesis also significantly decreased compared with other groups (p < 0.05). Supplementation of 50 μM Eicosapentaenoic acid showed higher quality blastocysts in terms of high cell numbers and low apoptosis when compared with other groups (p < 0.05), and synthesis ratio of E2/P4 also significantly increased compared with control group (p < 0.05). However, Supplementation of 100 μM Eicosapentaenoic acid showed high apoptosis when compared with other groups (p < 0.05), and synthesis ratio of 17β-estradiol/progesterone also significantly decreased compared with control group (p < 0.05). Our results indicated that supplementation with appropriate levels of Eicosapentaenoic acid beneficially affects the change of hormone synthesis for controlling oocyte maturation, leading to improved embryo quality. However, high doses of Eicosapentaenoic acid treatment results in detrimental effects.
Kim, D.H;Kang, H.G.;Kim, M.K.;Han, S.W.;Chi, H.J.;Lee, H.J.;Lee, H.T.;Chung, K.S.
Korean Journal of Animal Reproduction
/
v.24
no.4
/
pp.395-406
/
2000
The present study was conducted to examine the developmental capacity of mouse oocytes within prenatal follicles cultured various concentrations of FSH and LH and the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) and cytochrome P450 17 $\alpha$ -hydroxylase (P450)$_{17{\alpha}}$ mRNA, as luteinization and atretic marker, in these culture conditions. In addition, we investigated the concentrations of progesterone and testosterone in culture medium. The developmental potential up to blastocyst of the oocytes grown in vitro was higher in the FSH alone (30.2%) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (28.0%) groups than in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (22.0%). And the mean numbers of cell per blastocyst was higher in the FSH alone (50.9$\pm$26.1) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (51.0$\pm$21.1) groups when compared to the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (45.2$\pm$15.1). The expressions of P450scc and P450$_{17{\alpha}}$ mRNA in the oocyte -cumulus complexes were increased with increasing of LH concentration, and also the secretions of progesterone and testosterone were increased. Especially, in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group, the expression of P450scc and P450$_{17{\alpha}}$ were significantly increased, and the secretion of progesterone and testosterone were significantly increased. Therefore, these data show that gonadotrophins are essential for the in vitro culture of preantral follicles, but that increasing of LH concentration is reduced the developmental capacity of oocytes. The cause of these findings may be due to increasing of progesterone and testosterone secretion by the enhance of P450scc and P450$_{17{\alpha}}$ mRNA expressions, as markers of luteinization and atresia. Conclusively, this study suggest that supplementation of 100 $m\ell$U/$m\ell$ FSH or 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH may be optimal condition for the culture of mouse pre antral follicles.
Neosporosis is a widespread parasitic disease caused by Neospora caninum, an intracellular protozoan parasite. It causes economic losses due to reproductive failure. The potential relationship between pregnancy outcomes and levels of IFN${\gamma}$, hormones, and antibodies in naturally N. caninum-infected cows was examined in the blood samples collected every 2 or 4 weeks in 26 pregnant cows from 4 different farms. The mean S/P value of seropositive nonaborting animals (n = 14) reached peak levels 15 weeks prior to parturition, and declined thereafter to parturition. The S/P value 13 weeks prior to abortion in seropositive aborting cows (n=3) remained at high levels, and abortions occurred at 20 (142 days), 26 (185 days), and 28 weeks (199 days) after artificial insemination. IFN${\gamma}$ levels in the seropositive non-aborting group varied by individuals and gestational periods; IFN${\gamma}$ levels stayed at elevated levels or increased abruptly close to abortion in seropositive aborting cows. IFN${\gamma}$ level patterns in the seronegative group (n = 9) were similar to the seropositive non-aborting group, although IFN${\gamma}$ amounts were lower than the seropositive group. The mean progesterone levels in the seropositive non-aborting and seronegative groups decreased markedly 7 weeks prior to parturition. The mean progesterone levels 5 and 7 weeks prior to abortion were lower than the other groups 5 and 7 weeks prior to parturition. The mean 17${\beta}$-estradiol levels in the seropositive aborting cows increased close to abortion; the produced amounts were lower than those of seropositive non-aborting and seronegative groups close to parturition. These results suggested that lower levels of progesterone and ${\beta}$-estradiol in Neospora-infected cows may lead to increases in IFN${\gamma}$ production and in turn may result in abortion.
Kim, Sang-Don;Jee, Byung-Chul;Lee, Jung-Ryeol;Suh, Chang-Suk;Kim, Seok-Hyun;Moon, Shin-Yong
Clinical and Experimental Reproductive Medicine
/
v.37
no.1
/
pp.41-48
/
2010
Objectives: The aim of this study was to assess appropriate time to convert intramuscular progesterone support to oral administration for luteal phase support in in vitro fertilization (IVF). Methods: Seventy-six cycles of IVF in which fetal heart beat was identified after treatment were included. Patients underwent controlled ovarian hyperstimulation with GnRH agonist long protocol (n=7) or GnRH antagonist protocol (n=66). Cryopreserved embryo transfer was performed in three cycles. Luteal support was initiated by daily intramuscular injection of progesterone, and after confirmation of fetal heart beat, converted to oral micronized progesterone (Utrogestan, Laboratoires Besins International, France) 300 mg daily before or after 8 gestational weeks. The oral progesterone was continued for 11 weeks. Results: Overall clinical abortion rate was 3.9% (3/76) and mean time to conversion was $8^{+4}$ gestational weeks ($46{\pm}5.8$ days after oocytes retrieval). The abortion rate was 5.6% (1/17) and 3.4% (2/59) in patients with conversion before 7 weeks and after 8 weeks, respectively, which were not statistically significant (p=0.678). The miscarriages were occurred at $9^{+4}$ weeks, $11^{+3}$ weeks and $11^{+4}$ weeks. Conclusion: Sequential luteal support using intramuscular and oral progesterone yields a relatively low clinical abortion rate. If fetal heart beat confirmed, sequential regimen appears to be safe and convenient method to reduce patients' discomfort induced by multiple injections.
This study was performed to analyze the characterization of plasma hormonal levels during pregnancy in the Hanwoo recipients pregnant by artificial insemination (AI) or somatic cell nuclear transfer (SCNT) embryos. The synchronized recipients pregnant by SCNT embryos produced by Hanwoo fetal fibroblast cells (n=8) and by AI (control, n=5) were used. The plasma hormonal levels were measured by RIA (P4 and E2) and ELISA (cortisol), respectively. In control, the increase of E2 and the decrease of P4 were occurred immediately before the initiation of parturition. The expression pattern of plasma P4 was similar in both groups from 50 to 10 days before parturition, however, it did not decrease even at the expected date of labor in the SCNT recipients. The plasma cortisol was expressed a lower level during pregnancy in the SCNT recipients. But, the cortisol was increased in the cow aborted around 100 days of pregnancy (n=1). Based on these results, it can be postulated that the failure of the hormonal changes immediately before parturition in the SCNT recipients may be one of the most important reasons for a delayed parturition in clone calving.
There exists considerable evidence that steroid hormones are involved in the regulation of ovulation rate and oviductal development in poultry. However, the effect of steroid hormones on egg productivity of Korean Native Ogol Chicken (KNOC) has yet to be studied. Therefore, this study was performed to relate the expression of steroid hormones, especially progesterone ($P_4$) and estradiol ($E_2$), with egg productivity during the laying period. Egg production and egg weight of 70 KNOC were recorded from 20 to 60wk. Blood was taken every 10 wk and serum $P_4$ and $E_2$ were measured by radioimmunoassay. Based on egg productivity and steroid hormones levels up to 60 wk, chickens were divided into two groups, high and low. Compared to the low egg production group, a significantly higher expression of $P_4$ at 30 wk was detected in the high group. Moreover, egg production in the high $P_4$ group significantly differed from that in the low group at 30 wk. On the other hand, a Significant difference (p<0.05) in $E_2$ expression was found between high and low egg weight groups at 30 wk. Although a significant difference in egg weight between two groups by $E_2$ was not detected, the high $E_2$ group showed a higher level of egg weight than the low $E_2$ group except for 25 wk. In the comparison of ovary weight and small yellow follicle number, the group with high egg productivity and steroid concentration showed greater levels than the low group. Taken together, the results indicate that $P_4$ is related to egg productivity whereas expression of $E_2$ is associated with egg weight in KNOC.
Twenty-four Javanese thin-tail ewes (11, 9, and 4 ewes giving birth to 1, 2, and 3 lambs, respectively) with similar body weight and age at breeding were used to study serum progesterone concentrations during pregnancy, milk production during lactation, and mammary gland indices at the end of lactation (3 months postpartum). The results of the experiment showed that averages serum progesterone concentrations during pregnancy in the ewes giving birth to twin and triplet lambs were higher (p < 0.01) than those giving birth to a single lamb. Ewes giving birth to 3 lambs had higher (p < 0.01) mammary dry fat-free tissue (DFFT) (by 31 and 34%), DNA concentration (by 25 and 16%) and RNA concentration (by 29 and 16%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in mammary collagen, protein and glycogen concentrations at the end of lactation among litter sizes. Ewes giving birth to 3 lambs had higher (p < 0.01) total mammary DNA content (by 64 and 61%) and RNA content (by 69 and 53%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in total mammary collagen, protein and glycogen contents at the end of lactation among litter sizes. Even though ewes with higher litter size had numerically higher milk production, there was no significant difference in milk production per 4 h among litter sizes. The results of the experiment indicated that ewes having higher litter size had greater mammary cell number and synthetic activities at the end of lactation. The results suggested that ewes with higher progesterone concentrations and better developed mammary glands during pregnancy could maintain higher cell number and activities throughout lactation.
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