Interspecific cross-reactivity of deciduoma antiserum: interaction between mouse deciduoma and antiserum to rat deciduoma.
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Apoptotic cell death in artificially induced deciduoma of pseudopregnant mice.
Deciduoma induced by mechanical stimulation in pseudopregnant mice is similar to the decidua in normal pregnancy and it undergoes regression after a certain period. Therefore, we examined cell death in deciduomas which were induced by artificial stimulation. To analyze the regression mechanism of artificially induced deciduoma, DNA fragmentation, in situ 3'-DNA nick end labeling, and RT-PCR were performed on day 6 to 14 of pseudopregnancy. DNA fragmentation appeared on day 8 and it increased to day 10 of pseudopregnancy in the traumatized uterine horn. A large number of apoptotic cells were found on day 10 in the periphery of deciduoma at the antimesometrial side. Deciduoma underwent degeneration on day 11 of pseudopregnancy. Expression of tumor necrosis factor-alpha (TNF-alpha) mRNA was high on days 8 and 10, then decreased, whereas the expression increased again on day 14. TNF-alpha protein was expressed from day 8 to day 12, showing a peak expression on day 10 when deciduoma reached maximum weight. Serum progesterone level was high in the traumatized pseudopregnant mice on day 6, then it gradually decreased. Life span of deciduoma was prolonged 4 days more by daily injection of progesterone. A reduction in serum progesterone coincides with TNF-alpha increase, resulting in an increase of apoptotic deciduomal cells at the regression period, and that the life span of deciduoma is prolonged by additive supply of progesterone.
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Estrogen receptor in deciduoma cells separated by velocity sedimentation.
Nuclear estrogen receptor levels were measured in fractions of dispersed deciduoma cells separated according to size by velocity sedimentation at unit gravity. Previous studies demonstrated that velocity sedimentation separated larger fully differentiated polyploid deciduoma cells from smaller diploid cells. Since levels of estrogen receptor measured in homogenates of deciduoma tissue decreased during the growth and differentiation of the deciduoma, lower levels of estrogen receptor per cell were anticipated in polyploid deciduoma cells. Although levels of estrogen receptor decreased slightly in all fractions of deciduoma cells from days 7-8 of pseudopregnancy, the larger polyploid cells had higher levels of estrogen receptor than smaller cells on both days. These results indicate that differentiation of endometrial stromal cells during decidualization involves a decrease in receptor levels in all deciduomal cell types rather than a specific decrease in the receptor levels of only fully differentiated deciduoma cells. (Endocrinology 108: 484, 1981)
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A mechanistic assessment of 1,3-butadiene diepoxide-induced inhibition of uterine deciduoma proliferation in pseudopregnant rats.
Butadiene diepoxide (BDE), a reactive metabolite of 1,3-butadiene that is an important industrial chemical used in synthetic rubber production causes a dose-dependent inhibition of deciduoma development in pseudopregnant Sprague-Dawley rats. This study used 4 daily i.p. BDE doses of 0.20, 0.25, 0.30, 0.35, or 0.40 to characterize mechanisms that may be responsible for the antideciduoma effect. Pseudopregnant rats were treated either before (pseudopregnancy [PPG] days 1-4) or after (PPG days 5-9) deciduoma induction by endometrial trauma with a blunt needle. Animals were killed on PPG day 9 and evaluated for serum progesterone and endometrial protein and DNA. RT-PCR was used to measure message for estrogen receptor (ER) alpha and pituitary adenylate cyclase-activating polypeptide (PACAP). Substrate zymography and Western blotting were used respectively to measure matrix metalloproteinase (MMP)-9 and inducible nitric oxide synthase. The antideciduoma effects of BDE were associated with decreases in endometrial weight, protein, and DNA, with decreases in serum progesterone, and with decreases in PACAP message and MMP-9. A reduction in NOS was identified at the highest dose of BDE. Message for estrogen receptor (ER) alpha was not affected at any dose. We conclude that the reduction in decidual proliferation was direct and appeared to be associated with either 1) a decrease in the effectiveness of the deciduogenic stimulation and/or a weakened endometrial sensitivity to the stimulus; or 2) an effect on deciduoma development. Molecular mechanisms that apparently contributed to BDE inhibition of decidual metabolism included the synthesis of protein and DNA involved in decidual growth, the synthesis and activation of a matrix metalloproteinase for degradation of the extracellular matrix that is essential for tissue remodeling during deciduoma development, and the nitric oxide/nitric oxide synthase and pituitary adenylate cyclase-activating peptide systems that are involved in promoting vasodilation and increased vascular permeability to enhance the availability of substrates for maximal deciduoma growth. The ovotoxicity of BDE, which has previously been established, may indirectly affect decidual proliferation by reducing progesterone, the preeminent endocrine regulator of deciduoma development. The findings also suggest that BDE may possess no estrogenic action since it was associated with endometrial weight loss and unaltered levels of the estrogen receptor alpha mRNA expression.
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Relationship between deciduoma formation and uterine blood flow in the pseudopregnant rat.
The relationships between experimentally induced deciduoma formation, circulating estradiol (E) and progesterone (P) levels and alterations in uterine blood flow (UBF) were studied between days 4 and 15 of pseudopregnancy (PSP: day 0 = ovulation) in rats. Blood flow was measured with an electromagnetic blood flow monitor and serum analyzed for E and P levels by radioimmunoassay. Neither uterine trauma on day 4 of PSP nor the site of trauma had any direct influence on altering UBF. A dramatic increase in UBF occurred in response to stromal proliferation on days 5-6 and continued to remain above sham-operated control levels through day 9. These vascular changes correlated temporally with the day 9 peak in uterine weight resulting from deciduoma formation. Both uterine weight and vascular fluctuations in deciduoma-bearing rats were related to the cyclic changes in E/P ratios between days 5 and 8 of PSP. A decline in UBF preceded deciduoma regression between days 9 and 15 of PSP. These results suggest that an increase in UBF is causally associated with the formation and maintenance of deciduoma in the PSP rat and that deciduoma regression may result from a subsequent decline in UBF rates. Both factors may be directly dependent on fluctuating E/P ratios.
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Effect of ovariectomy in the early first half of the diestrus on induction or maintenance of canine deciduoma.
The effect of ovariectomy in the early first half of the diestrus was examined on the induction or maintenance of suture-induced canine deciduoma. Ovariectomy immediately, or some days, after the insertion of suture had no effect on the induction or maintenance of deciduoma. Even when ovariectomy was performed within 4 days before insertion, deciduoma could be induced in spite of there being no ovary. However, when ovariectomy was done 4 or more days before suture insertion, the rate of deciduoma was decreased or no deciduoma was induced. These results indicate that the influence of the ovary on the endometrium may persist for at least 4 days after ovariectomy. Ovariectomy after the suture insertion had few effects. It is suggested that canine uterine glands in the early first half of the diestrus maintain a certain degree of self-proliferative ability even after ovariectomy, and thus canine deciduoma is not as dependent on the ovary that of the rodentia.
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Progesterone regulation of progesterone receptor in mesometrial and antimesometrial deciduoma in pseudopregnant hamster.
Experiments were performed to examine the trophic effects of progesterone (P) on the progesterone receptor (PR) and the maintenance of the deciduoma. Deciduomal reactions in response to artificial stimuli were induced in hamsters' uteri on the 4th (D4) day of pseudopregnancy (PSP). On and after the 8th day, PSP hamsters received P supplement (2mg; s.c.) daily. Histological observation revealed that the life span of the deciduoma was partially prolonged. The maintenance and regression of the deciduoma was heterogeneous. P was able to maintain the morphology of the cells at the mesometrial site (MS) on D12, whereas cells at the antimesometrial site (AMS) regressed. The cytosol progesterone receptor (cPR) concentration in cells of the MS was higher than that of the AMS on D10. The cPR in MS and AMS decreased drastically on D12. The nuclear PR (nPR) remained at a higher concentration in both tissues on D10 followed by a precipitous decrease. The deciduomal nPR in the AMS decreased at a faster rate. By day 12, the nPR in the MS was much higher than that in the AMS. These data show that P acts as a trophic hormone of the deciduoma. The maintenance of deciduoma is closely related to the presence of PR. Other mechanisms may exist for the heterogeneous responses observed at the MS and AMS.
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Lipid peroxidation in the rat uterus during deciduoma induced cell differentiation.
The influence of deciduoma-induced differentiation on the lipid peroxidation in the rat uterus was investigated. The wet weight of uterus and its protein content increased during deciduoma progression. Content of the thiobarbituric acid reactive substances (TBARS) as well as lipid peroxidation induced by ascorbate and cumene hydroperoxide showed significant decreases during deciduoma growth. Restoration of normalcy was observed during regression. The activity of superoxide dismutase, an inhibitor of lipid peroxidation showed an opposite pattern namely increase during deciduoma development and decline during the regressive phase. We conclude that cell differentiation during deciduoma induction is accompanied by a temporary and reversible decrease in the peroxidative potential of the rat uterus.
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Expression of adenosine deaminase and 5'-nucleotidase in artificially induced deciduoma in rat and hamster.
Enzymes adenosine deaminase (ADA) and 5-nucleotidase (5-'NT) are known to play active role in tissue/cell proliferation and differentiation. To validate this the two enzymes were studied in artificially induced deciduoma of rat and hamster. The deciduoma was induced by traumatizing one of the uterine horns of progesterone primed animals. Non traumatized horn served as control. The animals were later maintained on progesterone, given alone (Gr.I) or conjointly with estrogen (Gr.II). The weight of each uterine horn was recorded to determine the formation of deciduoma. There was no marked difference between the weights of traumatized and control horn on day 2 post-traumatization (PT), but a progressive rise was noticed after this day in both species. The ADA activity however differed, day and species wise. While in the rats of Gr.I it was low in the traumatized horn on all the days, in the hamsters it was remarkably high from day 2 to 6 PT. In the rats of Gr.II also the activity though was low in the traumatized horn, but on day 2 and 4 only; on day 6 and 7 PT it increased markedly. In hamster, on the contrary, again the enzyme activity was remarkably high on all the three days. The 5'-NT activity, however, did not show any marked difference between the two horns under Gr.I and II in both species. It was rather high in the control horn of each group. The results suggest: (I) the progesterone alone though produces a significant rise in the uterine weight of traumatized horn in both species, the ADA activity increases only in hamster, (2) under the conjoint treatment also the enzyme activity remains high in hamster; and (3) the activity of enzyme 5'-NT does not alter during the deciduoma formation in both the species.
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Deciduoma formation in uterus of genetically mast cell-deficient W/Wv mice.
The role of mast cells in deciduoma formation in the uterus of mice was investigated by using genetically mast cell-deficient (WB x C57BL/6)F1-W/Wv (hereafter called WBB6F1-W/Wv) mice. Deciduoma formation occurred in castrated WBB6F1-W/Wv mice after estradiol-progesterone injection and traumatization in spite of their deficiency of mast cells. Injection of diphenhydramine, which blocks histamine receptors, inhibited deciduoma formation in WBB6F1-W/Wv mice as well as in congenic +/+ mice. THe uterus of WBB6F1-W/Wv mice contained about 10% of the amount of histamine found in the uterus of +/+ mice. These results suggest that mast cells are not essential for formation of deciduomata and that histamine originating from sources other than mast cells may be important in deciduoma formation.
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Characterization of deciduoma marker proteins in hamster uterus: detection in decidual cell cultures.
The objective of this study was first, to identify the proteins associated with decidualization of the hamster uterus by comparing the protein maps of decidualized and nondecidualized endometrium in vivo, and second, to determine whether decidual cell cultures produced these characteristic proteins. Decidualization was induced in one uterine horn, and the contralateral horn was not stimulated (control tissue). Animals were ovariectomized and a subcutaneous progesterone implant was used to maintain decidualization. Uterine proteins from nuclear and cytosol fractions were analyzed by two-dimensional electrophoresis using a highly sensitive protein staining technique. Analysis of nuclear extract and cytosol from decidualized and nondecidualized endometrium from Days 6, 7, and 8 of pseudopregnancy demonstrated the presence of 11 nuclear and five cytosolic deciduoma-associated proteins. Serum and erythrocyte proteins were identified by two-dimensional electrophoresis, and none of the 16 deciduoma-associated proteins was a serum or erythrocyte contaminant. Forty-eight-hour cultures of decidual cells harvested from Day 5 of pseudopregnancy produced all 16 of the deciduoma-associated proteins found in whole tissue in situ. Culture conditions minimized serum and erthrocyte contamination, enhancing the detection of deciduomal cell proteins. Four nuclear and two cytosolic proteins were considered deciduoma specific, i.e., they were not associated with cellular proliferation, as evidenced by their absence from cultures of rapidly dividing fetal hamster fibroblasts. Thus, these studies show that the detection of deciduomal proteins may be a useful criterion for the assessment of decidualization in vitro and in vivo.
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Induction of canine deciduoma in some reproductive stages with the different condition of corpora lutea.
Bitches were examined to see whether canine deciduoma could be induced at some reproductive stages with the different conditions of corpora lutea by inserting a silk suture into the uterine lumen. The bitches stimulated in the early and middle stages of diestrus or in unilateral pregnancy corresponding to these diestrous stages formed deciduoma at a high induction rate, however, no difference in the strength of decidual reaction between the pregnant and diestrous stages was recognized. On the other hand, no reaction could be seen in bitches in late diestrus, the late stage of unilateral pregnancy or the post partum repair phase in which stromal decidual cells similar to those of the rodentia can be seen. In already implanted uteri, however, no deciduoma was formed in the interplacental areas. Even though the corpora lutea were functional, new additional stimulations were not accepted at the interplacental area in which the uterine horn had already been influenced by fertilized ova. From these results, it was suggested that in the dog as well as the rodentia, the endometrium has to be under the influence of functional corpora lutea in order to form deciduoma.
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Changes of progesterone receptors in mesometrial and antimesometrial deciduoma after removal of fetus or placenta in unilateral pregnant hamster.
The effects of fetuses or placentas (with fetuses) removal on progesterone receptors (PR) in deciduomal tissues were investigated in unilateral pregnant (ULP) hamsters. Deciduomal reactions in response to artificial stimuli were induced on the tube-ligated uterine horns of ULP hamsters. Fetuses or placentas (with fetuses) were removed on day 8 of ULP, and the cytosolic and nuclear PR (cPR, and nPR, respectively) levels in deciduomal tissues at mesometrial (MS) and antimesometrial (AMS) sites were separately studied on day 10 and 12 of ULP. In the sham control group, cPR at the MS decreased with time, but at a slower rate than that at the AMS. The MS deciduoma contained higher cPR than the AMS deciduoma on day 12. Removal of fetuses did not cause any significant change of the decline rate, and the PR levels were similar to that of the control. The nPR at the MS and AMS also declined with time in all three groups. Removal of placentas induced drastic losses of cPR and nPR at the MS and AMS on day 10 and 12. Supplement of progesterone (P; 2 mg/animal/day) to the placenta-removed group restored the cPR somewhat at the MS site, but not back to the level of the control. The effect of P to restore nPR levels could be observed at the MS and AMS on day 10, but only at the MS on day 12. These data suggest that placental factor(s) other than P may play an important role on the maintenance of PR in deciduoma. Other cellular mechanisms may exist at the MS and AMS sites for the heterogeneous responses.
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Time-dependent relationship between the estrogen receptors and the matrix metalloproteinases following deciduoma induction in rats.
The purpose of this study was to investigate time-related interactions between the estrogen receptors, mediators of steroidal regulation of uterine growth, and an extracellular regulatory enzyme, the matrix metalloproteinases (MMPs) engaged in connective tissue degradation and remodeling that are fundamental to implantation and placentation. Pseudopregnant rats, in which the decidual response, the basis for decidualization, was surgically induced on day 4 of pseudopregnancy (PG), were sacrificed on PG days 3, 6, 9, and 15 for retrieval of uterine tissues for assays: the radioligand binding assay for the estrogen receptors and substrate zymography for the MMPs. Following increases on PG day 3, there were time-dependent decreases in the cytosolic low and high capacity estrogen receptors during deciduoma development (PG days 6-9) and regression (PG day 15) in both the endometrium and myometrium. Moreover, whereas the low capacity estrogen receptor levels were only slightly decreased (PG days 6-15), the high capacity receptors were reduced on day 6 (P < 0.001) and were completely diminished during PG days 9 and 15. In contrast, the MMPs (92 and 72 kDa) activities were increased from PG days 6-15 (P < 0.05) over the pre-decidual induction values on PG day 3 in both uterine compartments. The results suggest that deciduoma induction can modulate the concentration of cytosolic estrogen receptor subtypes and MMP activities in rats. The inverse time-dependent interrelationship between these cellular and extracellular components during deciduoma development and regression imply that the remodeling role of the MMPs may be enhanced by the reduced cytosolic estrogen receptor/estrogen action.
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Comparative study of biological properties of proteins synthetised in vitro by murine decidua and deciduoma.
Supernatants from short-term in vitro cultures of murine decidual tissue obtained from the uteri of pregnant mice on day 8 postcoïtum (decidua, in the presence of paternal antigens) or from pseudopregnant mice (deciduoma, without any histocompatibility antigens were assessed for their regulatory activity; both statuses (physiological and experimental) can only be developed under hormonal conditions. All these supernatants possess no complement inhibitory activity, but they markedly impair the generation of cytotoxic T cells (CTL) and suppress anti-sheep red blood cell antibody response. These effects seem essentially located in a single AcA 22 fraction of 60 KD that inhibits CTL, while the plaque-forming cell response is strongly stimulated. Electrophoretic and gel filtration profiles of supernatants from cultures of decidua and deciduoma seem similar. The results suggest that the factors synthetised by decidua or deciduoma are the same and are not induced by exposure to foreign histocompatibility antigens of the fetus. According to these results, it seems likely that decidualization represents a localized general mechanism that can be induced, under a particular hormonal background, by different stimuli (trauma) rather than being a specific response to a particular signal (blastocyst).
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Expression of rat prolactin-like protein B in deciduoma of pseudopregnant rat and in decidua during early pregnancy.
The rat PRL-like protein-B (rPLP-B) complementary DNA (cDNA), originally cloned from a late term placental library, hybridizes to transcripts in the deciduoma tissue artificially produced in pseudopregnant rats. The expression of rPLP-B in deciduoma is first observed 48 h (pseudopregnancy day 7) after the deciduogenic stimulus and increases to a maximum by 96-120 h (pseudopregnancy day 9-10). In situ hybridization studies show that rPLP-B hybridizes specifically to the antimesometrial cells of deciduoma tissue in day 9 pseudopregnant rats and to the homologous cells of the decidua that surround the embryo-trophoblastic structure at day 9 of pregnancy. A temporal study on the expression of rPLP-B during pregnancy shows that rPLP-B is concomitantly expressed by the decidual cells of maternal origin and the cytotrophoblast of fetal origin around day 13 of pregnancy.
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Histological variations of canine deciduoma induced in non pregnant horn at different stages of unilateral pregnancy.
Histological variations of canine deciduoma which was induced in the non pregnant horn at several stages of unilateral pregnancy were examined. In the first half of the unilateral pregnancy, deciduoma was characterized by the cystic glandular hyperplasia corresponding to each of the stages in normal early placentation. In the second half, deciduoma could not be induced and few histological reactions were recognized. The endometrium looked normal for late diestrus with no growth of the uterine glands. These differences might reflect the latent strength of the uterine glands to proliferate and dilate in the stimulated periods.
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Histological characteristics of canine deciduoma induced by intrauterine inoculation of E. coli suspension.
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.
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