Metabolism of arachidonic acid in sheep uterus: in vitro studies.

Arachidonic acid (AA) metabolism in the non-pregnant sheep uterus was studied in vitro using conventional chromatographic and HPLC techniques. High expression of both lipoxygenase (LOX) as well as cyclooxygenase (COX) enzymes and their activities was found in the uterine tissues. On incubation of uterine enymes with AA, the LOX products formed were identified as 5-hydroperoxyeicosatetraenoic acid (5-HPETE), 12- and 15-hydroxyeicosatetraenoic acids (12- and 15-HETEs), based on their separation on TLC and HPLC. By employing differential salt precipitation techniques, the LOXs generating products 5-HPETE (5-LOX), 12-HETE and 15-HETE (12- and 15-dual LOX) were isolated. Based on their analysis on TLC, the COX products formed were identified as prostaglandins - PGF2alpha and prostacyclin derivative 6-keto PGF1alpha. The study forms the first report on the comprehensive analysis on the metabolism of AA in sheep uterus in vitro via the LOX and COX pathways.

Myosin isoforms in uterine smooth muscle during pregnancy in rat.

Effects of uterine stretching and physiological hypertrophy on myosin isozyme were investigated in rat during pregnancy. Both nonpregnant and pregnant rat uteri express a single myosin band on native gels. Analysis of native myosin under denaturing conditions revealed two myosin heavy chains (MHCs) with molecular mass of 204 and 200 kDa respectively. Filamin, a 240 kDa protein co-electrophoreses with myosin on native gels. No correlation is found between regulatory myosin light chain phosphorylation and pattern of myosin isozymes or the MHC. The results suggest that uterine stretching and physiological hypertrophy during pregnancy do not induce any changes in uterine myosin isozyme.

Changes in uterine myosin isozymes from goat during early pregnancy.

Changes in the expression of native myosin, myosin heavy chains (MHCs) and myosin light chains (MLCs) were investigated in goat uterus during early pregnancy. Electrophoresis of native myosin under non dissociating conditions displayed two isozymes differing in their proportions during gestation. Three MHC isoforms were obtained following sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Whereas 204 kDa (a smooth muscle MHC) and 196 kDa (a non muscle MHC) were recorded in uterus from non pregnant goat, uterus of pregnant goat displayed a third 200 kDa smooth muscle MHC. Two conspicuous proteins (154 and 140 kDa respectively) in addition to MHCs were also obtained when the myofibrillar extracts were analysed either in the presence or absence of proteolytic inhibitors. Non pregnant goat uterus showed a basal level (ca.5%) of phosphorylation of regulatory myosin light chain. Uterine myosin from pregnant goat was recorded in a completely dephosphorylated state.

Prolactin suppression during pre and post-implantation periods on rat uterine glucosamine synthase activity.

Administration of bromocriptine (Bc), an ergot derivative having dopamine receptor agonist activity, to rats on day 1-5 of pregnancy prevented implantation of blastocysts and significantly suppressed uterine glucosamine 6-phosphate synthase activity. There was no effect on implantation or the enzyme activity when Bc was injected on day 7 or later of pregnancy. Injection of prolactin following Bc partially restored the enzyme activity and increased number of implantation sites. These results indicate that suppression of prolactin on day 1 to 5 of pregnancy causes failure of implantation. Bc on day 9 or later had no effect possibly due to the availability of placental LH/hCG to support the luteal cells.

Effects of centchroman, a synthetic estrogen antagonist on the fertility of female hamsters.

Centchroman (3, 4-trans-2, 2-dimethyl-3-phenyl-4-p-beta-pyrrolidinoethoxy-phenyl-7-methoxy-chroman) , a non-steroidal, estrogen antagonist, injected subcutaneously (2 mg/kg body wt) on days 1, 2 and 3 post-coitum in hamsters, prevented implantation in 70% of the animals. A significant decrease in the circulating levels of estradiol and progesterone was observed on day 4 post-coitum as compared to control animals following the treatment of centchroman. The activities of various lysosomal enzymes were also found diminished in the treated animals. This study shows that centchroman may act as an anti-implantation agent in hamsters indicating that estrogen plays a key role during the process of ovum implantation in this species.