• Korean Journal of Veterinary Research
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• v.31 no.2
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• pp.217-222
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• 1991

Most progesterone enzyme immunoassays(EIA) are used liquid phase double-antibody separation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of there several problems, we were prompted to develop an effective EIA system by the use of higher titer of progesterone antiserum free of anti-bovine serum albumin antibodies (anti-BSA). The results obtained were as follows. 1. The antibody of progesterone antiserum was high as $1.5{\times}10^5$. 2. Percent activity bound of progesterone antiserum was about 77 at a dilution to $5{\times}10^3$ times. 3. Progesterone antiserum was contained a large amount of anti-BSA antibodies. 4. The anti-BSA was completely absorbed by using of polymerised BSA. 5. The molecular weight of albumin polymer (polymerised BSA) obtained by using 2.5% glut. araldehyde was $5{\times}10^5$.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.1
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• pp.131-136
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• 1994

In an effort to confirm true oestrus and to detect early pregnancy in Zebu cows (Bos indicus), sequential blood and milk samples were collected at the day of imsemination (Day 0) and days 14, 20 and 24 after insemination. Progesterone was determined in skimmed milk and plasma by solid phase radioimmunoassay (RIA). Of the cows thought to be in oestrus plasma, (n = 46) and milk (n = 58) samples demonstrated low progesterone concentrations ($0.99{\pm}0.11$ and $2.02{\pm}0.14nmol/l$) in 42 (91%) and 52 (90%) cases respectively. Thirty two (76%) and 30 (71%) cows were thought to be pregnant by plasma progesterone RIA ($20.23{\pm}1.03$ and $20.48{\pm}1.01nmol/l$) at days 20 and 24 respectively. At the same period, 40 (77%) and 37 (71%) cows were thought to be pregnant by milk progesterone RIA ($27.82{\pm}1.28$and $28.02{\pm}1.27nmol/l$). Assuming 100% accuracy for rectal examination of pregnancy diagnosis between days 60-65 postservice, the RIA was found to be 84% and 90% accurate for plasma and 84% and 92% accurate for milk at day 20 and 24, respectively. All cows thought to be non pregnant by progesterone measurement were correctly diagnosed. Progesterone assay at 24 days after oestrus may therefore be accurate for early diagnosis of pregnancy in Zebu cows.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.306-311
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• 1993

The effects of methanol. used as a solvent for the hydrophobic substrate progesterone. on the morphology of Rhizopus nigricans and 11$\alpha$-hydroxylation of progesterone was investigated. The methanol tolerance of the 11$\alpha$-hydroxylase system in polyacrylamide immobilized R. nigricans mycelia as well as in free mycelia has been induced by adding various unsaturated fatty acids. biotin and ions into the cultivation medium. Immobilization of the cell seemed to protect the cells from denaturation by methanol. It gave higher reaction rate of progesterone than the free mycelia in the presence of methanol.500 $\mu$g/l of biotin was found to be the most effective induction agent for the methanol tolerance among tested chemicals. R. nixricans cells sustained its enzymatic activity at higher methanol concentrations as a result of accumulation of unsaturated fatty acids. especially oleic acid. in the membrane phospholipid.

• The Korean Journal of Mycology
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• v.11 no.3
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• pp.115-119
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• 1983

The bioconversion of progesterone by Aspergillus phoenicis has been studied. The metabolism in the conditions of experiment gave $11{\alpha}-hydroxyprogesterone$ as main product. The concentration of $11{\alpha}-hydroxyprogesterone$ increased monotonically and leveled off after 40 hours of incubation. Addition of glucose into the medium reduced considerably the time for attaining limiting concentration of $11{\alpha}-hydroxyprogesterone$. The increase in initial progesterone concentration did not affected the percentage of conversion nor the time required for termination of the reaction. But it could not be represented as first order reaction with respect to progesterone concentration. The degree of inhibition of enzymes by organic solvents depended upon the concentration of solvents. At low solvent concentration, acetone proved to yield the highest conversion.

• Animal cells and systems
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• v.5 no.1
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• pp.51-57
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• 2001

We have previously shown that oocyte maturation is induced by an immobilized progesterone, progesterone-3-carboxymethyloxime - bovine serum albumin conjugate (P-BSA) in Rana dybowskii. In this study, we confirmed the maturation inducing activity of P-BSA on Xenopus oocyte and examined the binding character of the immobilized progesterone on the surface of Xenopus oocytes after removal of the vitelline layer. P-BSA induced maturation of Xenopus oocytes but E-BSA failed to do so as observed in Rana. Binding of the immobilized progesterone, fluorescein isothiocyanate-labeled progesterone-3-0-carboxymethyloxime-BSA (P-BSA-FITC) on the devitellined oocytes surface was examined by fluorescence confocal microscopy. The binding affinity of P-BSA-FITC to the devitellined oocyte was higher than that of estrogen-BSA-FITC (E-BSA-FITC) or testosterone-BSA-FITC (T-BSA-FITC). The binding disappeared in the presence of excess free progesterone but not in the presence of free estrogen. Maximum binding occurred after two-hours of incubation with P-BSA-FITC at pH 7.5. Stronger binding occurred in oocytes at stage Vl than stage IV, and in vitro treatment of hCG enhanced the binding. Taken together, these results suggest that a specific receptor for progesterone exists on the plasma membrane of Xenopus oocytes and that progesterone acts initially on this putative receptors and triggers generation of membrane-mediated second messengers during the early stage of oocyte maturation In amphibians.

• Development and Reproduction
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• v.11 no.1
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• pp.21-29
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• 2007

Previously, we found progesterone receptor membrane component 2 (pgrmc2) was highly expressed in germinal vesicle (GV) stage oocytes. The present study was conducted to characterize the expression of pgrmc2, as well as pgrmc1, in the mouse gonads and embryos according to their developmental stages. We found that these membrane components were expressed in ovaries, testes, and embryos at various developmental stages in addition to oocytes. Progesterone-3-O-carboxymethyl oxime-BSA-fluorescein isothiocyanate (P4-BSA-FITC) was applied to visualize the presence of the progesterone receptor on mouse oocyte membrane, and we confirmed that immobilized progesterone is localized at surface of the oocyte. This is, at our knowledge, the first report regarding the expression of membrane component of progesterone receptor in the mouse oocytes, embryos, and gonads. The function and signal transduction pathway of progesterone receptor membrane components in oocytes requires further studies.

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.429-434
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• 1992

This study was carried out to develop an effective liquid-phase double antibody enzyme immunoassay for determining of progesterone. The optimum conditions of assay system, 1st and 2nd antibodies, enzyme conjugate, and time reaction were invested. The bovine plasma progesterone level in dairy cattle and korean native bulls were also analyzed. The results obtained were as follows; 1. The reproducibility of petroleum ether was superior to that of ethyl ether as extract solvent of progesterone in plasma. 2. The optimum dilution rate of 1st and 2nd antibody was 30,000 and 10 times, respectively. Affer the reaction of enzyme conjugate to progesterone 1st antibody, and then 2nd antibody competition reaction was enough for over 1hr. 3. Average plasma progesterone level in 4 pregnant and 9 nonpregnant Holstein was $2.5{\pm}0.5$ and $0.7{\pm}0.2ng/m{\ell}$, respectively. Average plasma progesterone level of 10 Korean native bulls was $0.1{\pm}0.001ng/m{\ell}$ From these results, by using liquid phase double antibody enzyme immunoassay for progesterone is applicable to detect of early pregnancy diagnosis, factorial analysis of reproductive disorder, and also reproductive physiological function such as monitoring of cyclicity during the post-partum period.

• The Korean Journal of Zoology
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• v.28 no.2
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• pp.99-107
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• 1985

In order to investigate the synergistic effect of dbcAMP and progesterone which are known as the agents to inhibit maturation of mammalian oocytes in vitro, the present studies were done and the results were obtained as follow: 1) if 10 $\\mug$/ml of dbcAMP or 2 $\\mug$/ml of progesterone was given into the medium, each of the agents at the concentration above did not give any inhibitory effect on the maturation of the mouse oocytes in vitro; 2) however, when the two agents at the concentration shown above were given together into the medium, the mouse oocytes were arrested at GV stage; and 3) the oocytes, precultured in the medium containing two agnts at the same concentration as above for four hours, resumed their maturation division upon transfer to the plain medium for the extended culture. Thus, it was found that dbcAMP and progesterone were capable to suppress the maturation of mouse oocytes at the suboptimal concentration when they were given together, and such inhibitory effect was reversible.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.281-287
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• 1990

This study was conducted to find out the changes of ovarian, placental and fetal weights and periods of pregnancy in rats implanted with progesterone-tube during the reproductive stages. One hundred and thirty-four mature rats, 10~13 weeks old, were offered for this experiment. The animals, which were implanted with silicon tubes filled with progesterone on day 15 of pregnancy, were sacrified at 18, 20, 21 and 22 days of pregnancy. The changes of ovarian, placental and fetal weights and the number of fetuses during late pregnancy were recorded. The results obtained were summarized as follows : 1. After progesterone-tube implantation, ovarian weight reached to a peak value of 92.0$\pm$0.9mg at 20 days of pregnancy, there after decreased significantly to 79.5$\pm$7.6 and 68.26$\pm$4.2mg at 20 and 22 days of pregnancy(P<0.01). 2. The placental weight increased rapidly during 15~18 days of pregnancy in control and progesterone treated rats. A peak value of 447.78$\pm$20.9mg was shown at 20 days of pregnancy after progesterone-tube implantation, and in control rats the value decreased significantly to 419.42$\pm$11.6 and 404.1$\pm$29.3mg at 20 and 21 days of pregnancy(P<0.01). 3. The fetal weights was not shown any significant differences between control and progesterone-tube implanted rats. 4. The number of fetuses in control rats were 14.75$\pm$0.4 at 8~10 days of pregnancy and 13.5$\pm$0.3 and 13.25$\pm$0.4 at 12 and 20 days of pregnancy. 5. The significant difference in periods of pregnancy was appeared between progesterone-tube implanted(27.3$\pm$0.3 days) and control(22.1$\pm$0.3 days)rats(P<0.01).

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.57-65
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• 1990

These experiment were carried out ; (1) to investigate the changes of progesterone in plasma, whole milk and skim milk during oestrus cycle and pregnancy and ; (20) to evaluate a chemiluminescence Immunoassay(CIA) as an alternative method by measuring the progesterone concentration is skim milk by RIA and CIA. The results obtained in these experiments were summerized as follows ; 1. Plasma progesterone levels in non-pregnant Holstein during oestrus cycle were relatively low until day 8 after oestrus. And then, the progesterone level began to increase and reached a peak with 6.3ng/ml on day 14 and then declined repidly to 1.5ng/ml and 2.2ng/ml on day 18 and 20, respectively. 2. Whole milk progesterone level in pregnant Holstein increased from 1.0ng/ml on oestrus to 16.0ng/ml on day 8 and then remained from 11.0ng/ml on day 10 to 22.0ng/ml on day 22. 3. In non-pregnant Holstein, whole milk pregesterone lev디 was 1.5ng/ml on oestrus and began to increase rapidly from day 6 after oestrus and exhibited a ranged of levels, 17.8~20.0ng/ml from day 6 to day 16 after oestrus. 4. Skim milk progesterone levels in pregnant Holstein were a range of 130~490pg/ml at the time of estrus and began to increase continually till then showing constant levels ranging from 1300pg/ml on day 10 to 1650pg/ml on day 22. 5. In non-pregnant Holstein, skim milk progesterone level was 160pg/ml on oestrus and began to increase from 190pg/ml on day 2 after oestrus to day 8 and then keep constant levels ran ging from 1050 to 1300pg/ml from day 8 to day 16 and then decreased to 240~450pg/ml from day 18 to day 22 after oestrus. 6. The results obtained from CIA for the analysis of skim milk progesterone were in good agreement with the values derived by RIA.(r=0.914)