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Life Sciences Jan 2001We determined whether fragmentation of genomic DNA, apoptosis, occurs during deciduomal regression in pseudopregnant hamsters and the effect of progesterone on the...
We determined whether fragmentation of genomic DNA, apoptosis, occurs during deciduomal regression in pseudopregnant hamsters and the effect of progesterone on the apoptotic processes. Artificially induced deciduoma were obtained on different days of pseudopregnancy and separated into mesometrial and antimesometrial tissues. The deciduomal cell cycle progression and population profiles of both sides were compared by flow cytometry. The proportion of sub-G1 peak, which was correlated with the apoptotic cells, were about 10% on day 8 and reached to 40% in both tissues on day 10. Exogenous progesterone treatment by daily injection (2 mg; s.c.) on and after day 8 reduced the percentage of low molecular weight DNA in both tissues on day 10 and day 12 as compared to the nontreated control one, respectively. The appearance of DNA ladder was also delayed at least 24 h by progesterone administration. The intensity of DNA fragmentation was more pronounced in antimesometrial deciduoma. In situ 3'-end labeling of apoptotic cells further substantiated the apoptotic process. The apoptotic cells first appeared in the luminal region in antimesometrial deciduoma on day 8 and spreaded all over the entire deciduomal tissue on day 10. Progesterone treatment stimulated deciduomal proliferating cell nuclear antigen (PCNA) expression, maintained deciduoma until day 14 and retarded the differentiation and regeneration of the uterine epithelium.
Topics: Animals; Apoptosis; Cell Division; Cricetinae; DNA; DNA Fragmentation; Decidua; Female; Flow Cytometry; In Situ Nick-End Labeling; Mesocricetus; Molecular Weight; Progesterone; Proliferating Cell Nuclear Antigen; Pseudopregnancy
PubMed: 11205872
DOI: 10.1016/s0024-3205(00)00984-x -
Molecular Reproduction and Development Feb 2001To investigate the molecular mechanisms of implantation, we constructed a complementary DNA library of mouse uterus enriched with pregnancy-induced genes by subtractive...
To investigate the molecular mechanisms of implantation, we constructed a complementary DNA library of mouse uterus enriched with pregnancy-induced genes by subtractive hybridization and polymerase chain reaction. One of the isolated clones was a part of complementary DNA for the Ly-6A/E. Ly-6A/E is reported to be differentially expressed on hematopoietic stem cells and some lymphoid and nonlymphoid tissues, mediate cell-cell adhesion on lymphoid cells, and associate with cell proliferation and angiogenesis of tumor cells. Northern blot, in situ hybridization, and immunohistochemical analyses demonstrated that the Ly-6A/E mRNA and protein were expressed in the endometrial epithelial cells as well as myometrial cells and vascular endothelial cells in the uterus of nonpregnant mouse. The expression was downregulated in luminal epithelial cells during pregnancy days 1-5, while it was upregulated in decidualized stromal cells around the implanted embryo at the time of implantation. The signals were primarily localized in stromal cells at the mesometrial pole on day 9. The increased expression was also observed in stromal cells of the embryo-transferred uterus and artificially-induced deciduoma, indicating that the expression of Ly-6A/E in the endometrial cells is concurrent with decidualization. These findings suggest that Ly-6A/E plays a role in embryo implantation.
Topics: Animals; Antigens, Ly; Blotting, Northern; Cloning, Molecular; DNA, Complementary; Embryo Implantation; Endometrium; Female; Gene Expression; In Situ Hybridization; Male; Membrane Proteins; Mice; Microscopy, Fluorescence; Pregnancy; RNA, Messenger; Stromal Cells; Uterus
PubMed: 11139227
DOI: 10.1002/1098-2795(200102)58:2<159::AID-MRD4>3.0.CO;2-T -
Journal of Applied Toxicology : JAT 2000The cytotoxicity of hydroxyurea (HU), currently used to combat various cancers, sickle cell anemia and human immunodeficiency infection, was assessed by exposing...
The cytotoxicity of hydroxyurea (HU), currently used to combat various cancers, sickle cell anemia and human immunodeficiency infection, was assessed by exposing decidualized and pregnant uteri of Sprague-Dawley rats to this drug. Consecutive daily doses of HU (500 mg/kg(-1)) for 4 days were injected subcutaneously during decidualization when proliferation of the deciduoma was biochemically analyzed on pseudopregnancy day 9, or injected intraperitoneally during pregnancy when uterine developmental processes were evaluated on gestation day 16. Hydroxyurea displayed prominent antiproliferative effects on decidual growth. These actions were comparable to significantly impaired (P<0.001) developmental responses (increases in post-implantation losses, in resorbed fetuses and in reduced fetal and placental weights) during pregnancy. The cellular components inhibited by HU were DNA, protein, nitric oxide synthase, a matrix metalloproteinase and decidual prolactin-related protein mRNA (P<0.05). Steroid-related endocrine events (serum progesterone concentrations, estrogen receptor and mRNA levels) were unaffected by HU, implying direct cellular action by the drug. Interestingly, endometrial alkaline phosphatase bioactivity was enhanced by HU (P<0.05). Subsequently, the reproductive toxicity of HU was apparently related to mitogenic and differentiation-induced endometrial cellular activities.
Topics: Animals; Blotting, Western; Decidua; Enzyme Inhibitors; Female; Hydroxyurea; Matrix Metalloproteinases; Nitric Oxide Synthase; Pregnancy; Pseudopregnancy; RNA, Messenger; Rats; Rats, Sprague-Dawley; Receptors, Estrogen; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleotide Reductases; Uterus
PubMed: 11139171
DOI: 10.1002/1099-1263(200009/10)20:5<407::AID-JAT704>3.0.CO;2-T -
Biology of Reproduction Dec 2000Tenascin-C (TN-C), an extracellular matrix glycoprotein, is known to be expressed in uterine stroma in the peri-implantation period. Examination of the spatiotemporal...
Tenascin-C (TN-C), an extracellular matrix glycoprotein, is known to be expressed in uterine stroma in the peri-implantation period. Examination of the spatiotemporal pattern during early pregnancy using immunohistochemistry and in situ hybridization revealed TN-C expression in the stroma beneath the luminal epithelia of the murine endometrium on Days 0 and 1 of pregnancy, subsequent disappearance, and reappearance on Day 4. After decidualization, tissue around the deciduoma was positive. In situ hybridization demonstrated TN-C production by the stromal cells adjacent to the epithelia. To investigate the regulation of TN-C expression in vitro, murine uterine stromal and epithelial cells were isolated and cultured. Addition of interleukin-1 alpha (IL-1 alpha) and prostaglandin E(2) (PGE(2)) and F(2 alpha) (PGF(2 alpha)) induced TN-C expression in the stromal cells at both protein and mRNA levels, while the sex steroid hormones, progesterone and ss-estradiol, exerted little effect. Immunohistochemistry using anti-IL-1 alpha antibody showed epithelial cells to be positive on Days 2-4 of pregnancy, and addition of progesterone but not ss-estradiol enhanced IL-1 alpha expression in epithelial cells in vitro. In a culture insert system, TN-C expression by stromal cells cocultured with epithelial cells was induced by addition of progesterone alone that was blocked by additions of anti-IL-1 alpha antibody. Collectively, these findings indicate that TN-C expression in the preimplantation period is under the control of progesterone, but not directly, possibly by the paracrine and autocrine intervention of IL-1 alpha secreted by epithelial cells and PGE(2) and PGF(2 alpha) secreted by stromal cells.
Topics: Animals; Cells, Cultured; Coculture Techniques; DNA Primers; Digoxigenin; Dinoprost; Dinoprostone; Endometrium; Epithelial Cells; Female; Immunohistochemistry; In Situ Hybridization; Interleukin-1; Mice; Mice, Inbred C3H; Pregnancy; Pregnancy, Animal; Reverse Transcriptase Polymerase Chain Reaction; Stromal Cells; Tenascin; Uterus
PubMed: 11090440
DOI: 10.1095/biolreprod63.6.1713 -
Biochemical and Biophysical Research... Jul 2000Indoleamine 2,3-dioxygenase (IDO) is expressed in trophoblasts and defends the conceptus against rejection by reducing the tryptophan level and suppressing the T cell...
Indoleamine 2,3-dioxygenase (IDO) is expressed in trophoblasts and defends the conceptus against rejection by reducing the tryptophan level and suppressing the T cell activity. We isolated a cDNA for tryptophan 2,3-dioxygenase (TDO), another key catabolizing enzyme of tryptophan, from a mouse uterus cDNA library enriched with pregnancy-induced genes. Northern blot and in situ hybridization analyses demonstrated that the TDO mRNA was induced in the decidualized stromal cells around the implanted embryo at the time of implantation. The expression was then upregulated and primarily localized at the mesometrial decidua. TDO mRNA was induced by deciduoma formation as well as embryo transfer but not by ovarian steroid hormones. These findings demonstrated that TDO is induced in the endometrial stromal cells concomitant with decidualization and suggested its involvement in the implantation process by regulating the tryptophan level at the implantation site.
Topics: Animals; Blotting, Northern; DNA, Complementary; Embryo Implantation; Embryo, Mammalian; Endometrium; Enzyme Induction; Estradiol; Female; Gene Library; In Situ Hybridization; Indoleamine-Pyrrole 2,3,-Dioxygenase; Male; Mice; Ovariectomy; Progesterone; RNA, Messenger; Transplantation, Homologous; Tryptophan; Tryptophan Oxygenase; Up-Regulation
PubMed: 10903913
DOI: 10.1006/bbrc.2000.3115 -
Indian Journal of Experimental Biology Aug 1999The artificially induced rat deciduoma serves as a model to study cellular changes associated with implantation in the endometrium. The stromal cells differentiate to...
The artificially induced rat deciduoma serves as a model to study cellular changes associated with implantation in the endometrium. The stromal cells differentiate to form two types of decidual cells and are restricted to specific anatomical sites of the uterus. Programmed cell death starts in the antimesometrial area and expression of glutathione-S-transferase, an antioxidant enzyme, enhances in these cells as the deciduoma enters the regressive phase. The enzyme activity is significantly high compared with that of mesometrial decidual cells. Similarly, lipid peroxide content of antimesometrial decidual cells is high during this phase. DNA fragmentation, a feature of cells undergoing programmed cell death, is initiated in the antimesometrial area during regression of deciduoma.
Topics: Animals; Apoptosis; Placenta; Rats; Rats, Wistar
PubMed: 10709322
DOI: No ID Found -
Indian Journal of Experimental Biology Apr 1999A pure compound, isolated from ethyl acetate extract (root) of D. mitis D. Don, prevented pregnancy by 100% in adult female hamster but partially in rat when...
A pure compound, isolated from ethyl acetate extract (root) of D. mitis D. Don, prevented pregnancy by 100% in adult female hamster but partially in rat when administered orally on Days 1-7 and 1-10 post-coitum respectively. The effective dose in both species was 150 mg/kg. Using uterine wet weight in ovariectomized immature rat as bioassay method, the compound was found to be devoid of estrogenic and antiestrogenic property. On examination for progestational and antiprogestational activity, using trauma-induced deciduoma formation in immature rat uterus as end points, the compound (per se) did not show the former activity but in a conjoint treatment with progesterone it augmented the action of latter. The compound was assumed to act by potentiating progesterone biosynthesis, the excess of which might be the cause for interruption of pregnancy in hamster. This is the first study to report contraceptive efficacy and mode of its action at the uterine level.
Topics: Animals; Contraceptives, Oral; Cricetinae; Female; Magnoliopsida; Male; Mesocricetus; Plant Extracts; Plants, Medicinal; Pregnancy; Progesterone; Rats; Rats, Sprague-Dawley; Uterus
PubMed: 10641178
DOI: No ID Found -
Gynecological Endocrinology : the... Oct 1999Trimegestone is a novel norpregnane progestin that is being developed, in combination with estradiol, for the treatment of menopausal symptoms. The pharmacological... (Comparative Study)
Comparative Study
Trimegestone is a novel norpregnane progestin that is being developed, in combination with estradiol, for the treatment of menopausal symptoms. The pharmacological characteristics of trimegestone have been evaluated in both in vitro and in vivo test systems, and compared with reference progestins. Interaction with hormonal steroid receptors from animal tissues and with human recombinant receptors in vitro has demonstrated that trimegestone has a high specificity and potency for the progesterone receptor, no affinity for the estrogen receptor, and weak affinity for androgen, glucocorticoid and mineralocorticoid receptors. With respect to progestomimetic activity in vivo, trimegestone was more potent than reference progestins in the endometrial transformation test in the rabbit, preventing the uterotrophic effect of estradiol in the immature mouse bioassay, and had more effect on traumatic deciduoma formation and greater oral antiovulatory activity in the rat. In vivo, trimegestone effectively maintained pregnancy in the rat, but was devoid of any uterotrophic activity. Trimegestone showed no androgenic, glucocorticoid, antiglucocorticoid or mineralocorticoid activity, but did show some antiandrogenic and antimineralocorticoid activity at higher doses. Administration of trimegestone to ovariectomized rats, in combination with estradiol, inhibited the uterotrophic effects of estradiol. At doses up to 1 mg/kg intravenously and 30 mg/kg orally, trimegestone was not associated with any unwanted pharmacological effects. Overall, the results show trimegestone to have a favorable pharmacological profile with potent progestomimetic activity.
Topics: Administration, Oral; Administration, Topical; Animals; Binding, Competitive; Endometrium; Female; Humans; Injections, Subcutaneous; Levonorgestrel; Medroxyprogesterone; Mice; Norethindrone; Norpregnenes; Progesterone Congeners; Promegestone; Rabbits; Rats; Rats, Sprague-Dawley; Receptors, Androgen; Receptors, Estrogen; Receptors, Glucocorticoid; Receptors, Mineralocorticoid; Receptors, Progesterone; Specific Pathogen-Free Organisms; Uterus
PubMed: 10599548
DOI: 10.3109/09513599909167574 -
European Journal of Endocrinology Oct 1999CDRI-85/287 is an anti-estrogen and interferes with decidualization in the rat uterus. In this study, uterine estrogen receptor (ER) and progesterone receptor (PR)...
CDRI-85/287 is an anti-estrogen and interferes with decidualization in the rat uterus. In this study, uterine estrogen receptor (ER) and progesterone receptor (PR) levels were determined during the inhibition of decidualization. The effect of 85/287 on uterine ornithine decarboxylate (ODC) activity (a marker enzyme for decidualization) was also studied, using immature ovariectomized rats divided into four different groups: control, 2.5mg/kg 85/287 only; 1 microg estradiol only; and 85/287 + estradiol. Pseudopregnant rats were administered 85/287 (2.5mg/kg p.o.) on day 3 post-coitum. Deciduoma was induced in one of the uterine horns on day 4 and animals were autopsied 18h post-traumatization. Both ERs and PRs showed an increase in traumatized horns compared with non-traumatized. In the 85/287-treated uterus, there was a reduction in cytosolic ERs in both traumatized horns. However, nuclear ER and PR levels increased in both horns under the influence of 85/287. Similarly, in a tamoxifen (0.03mg/kg)-treated group a decline was noticed in cytosolic ER with a mild increase in nuclear PR. Total ER content, expressed per 100 microg DNA, showed a decline in 85/287- or tamoxifen-treated rats. However, no significant alterations were observed in total PR levels in non-traumatized horns. In an immature rat model, 85/287 caused a significant (>50%) reduction in estradiol-induced ODC activity. These findings suggest that the decidualization inhibitory activity of 85/287 may be attributed to inhibition of certain timed biochemical events and genomic/non-genomic actions of estrogens in the rat uterus.
Topics: Animals; Benzopyrans; Decidua; Estrogen Antagonists; Female; Ornithine Decarboxylase; Piperidines; Rats; Rats, Sprague-Dawley; Receptors, Estrogen; Receptors, Progesterone; Receptors, Steroid; Uterus
PubMed: 10526259
DOI: 10.1530/eje.0.1410426 -
The Journal of Veterinary Medical... Apr 1999Canine deciduoma could be induced in the diestrous uterus by an intrauterine inoculation of a culture suspension of E. coli originally isolated from naturally occurring...
Canine deciduoma could be induced in the diestrous uterus by an intrauterine inoculation of a culture suspension of E. coli originally isolated from naturally occurring canine pyometra. These deciduomas had the same histological findings as those of naturally occurring canine pyometra with so called "Swiss cheese endometrium". This suggests a possibility that the canine pyometra is a kind of naturally occurring decidual reaction (deciduoma) induced by one of several triggers such as bacterial infection.
Topics: Animals; Decidua; Dog Diseases; Dogs; Endometritis; Escherichia coli Infections; Female; Pregnancy
PubMed: 10342298
DOI: 10.1292/jvms.61.433