Changes in the specific cytochemical properties of rat decidual cells during differentiation

The antimesometrial part of rat's decidua of 8-9 day of gestation was divided into three parts: epithelial, transition and basal zones. Cells of either zone have their own morphological and cytochemical features. Cells of the epithelial zone are characterized by synthesis of tissue specific antigens and fat (fatty acids). Cells of the transition zone synthesize glycogen. Acid mucopolysaccharides are located in the basal zone only. Decidual cells do not synthesize cholesterol. Con A receptors are localized on the surface of cells of the basal and transition zones and disappear from the surface of epithelial zone cells. It is concluded that differentiation of large decidual cells of the antimesometrial part of rat's decidua are accompanied by a significant change in cytochemical features of cell precursors the synthesis of acid mucopolysaccharides and glycogen stops, while tissue specific antigens and fat (fatty acids) start their syntheses, Con A receptor disappear.     

Progesterone withdrawal and RU-486 treatment stimulate apoptosis in specific uterine decidual cells

Progesterone secretion is required for the growth and differentiation of endometrial stromal cells to form decidual cells. For many cells where a growth factor supports cell growth and proliferation, withdrawal of the growth factor initiates apoptosis. This study determined the time course and tissue location of apoptosis in deciduomal tissue after withdrawal of progesterone or injection of the antiprogestin, RU-486. Total DNA was isolated from decidual tissues at intervals after experimental treatments and separated electrophoretically. Internucleosomal DNA fragmentation characteristic of apoptosis was measured by quantitating levels of the 200 bp fragment. Apoptotic cells in tissue sections were detected by direct immunoperoxidase detection of digoxigenin-labeled DNA. Decidual apoptosis reached maximal levels at 12 h after withdrawal of progesterone or injection of RU-486. Increased concentrations of apoptotic cells were observed at the periphery of the growing deciduoma and in the antimesometrial deciduoma near the luminal epithelium after both treatments. These results suggest the withdrawal of progestin initiates apoptosis in cells at the early stages of decidualization.     

Role of platelet-activating factor (PAF) in the initiation of the decidual reaction in the rat

Injection of PAF into the left uterine horn induced a dose-dependent decidua-like reaction in the pseudopregnant rat. This reaction was maximal when PAF was injected at Day 5 of pseudopregnancy and was blocked by the specific PAF antagonist, BN 52021. BN 52021 did not interfere with the decidual reaction induced by prostaglandin E-2 or insertion of a cotton thread in the uterine horn. In contrast, a decidua-like reaction was not evoked by the inactive lyso-PAF, demonstrating the specificity of the action of PAF. The decidua-like reaction induced by PAF involves the generation of cyclooxygenase metabolites of arachidonic acid since it was inhibited by indomethacin. The histological alterations induced by PAF were similar to those observed after embryo implantation, strengthening the postulate for a role of the autacoid in the early stages of pregnancy.     

Synergistic induction of apoptosis in primary rat decidual cells by INF-gamma and TNF

In the rat, in response to blastocyst implantation, stromal cells of the endometrium proliferate and differentiate into decidual cells, forming the decidua. After reaching its maximum development, the decidua undergoes regression. This phenomenon appears to be due to an active process involving apoptosis. As there is sparse knowledge concerning the mechanisms of induction of decidual cell death, the potential role of cytokines present in the uterine environment during pregnancy, such as tumor necrosis factor (TNF) and interferon-gamma (INF-gamma) was explored in primary cultures of rat decidual cells. The effects of these factors upon cellular viability, nuclear morphologic alterations, expression, and enzymatic activities of the effector caspases-3/7 were evaluated. The results obtained demonstrated that in contrast to TNF, which did not induce any alteration, INF-gamma and in association with TNF caused a decrease in cell viability and an increase in the appearance of apoptotic bodies in a time-dependent manner that was augmented in the co-presence of TNF. An increase in caspase-3/7 activities after 12 hr of TNF/INF-gamma treatment was also observed. These findings suggest that INF-gamma expressed in the uterine environment may play an important role in regulating apoptosis through potential synergistic mechanisms with TNF and thereby modulate decidual stability and regression during pregnancy.     

Ontogeny of adenosine deaminase in developing trophoblast and decidual cells of rat and hamster

The enzyme adenosine deaminase (ADA) is expressed at high level in the tissue of foeto-maternal interface during early pregnancy. As the main constituents of this interface are trophoblast (TR) and decidual cells (DC), the enzyme was estimated in isolated TR and DC to determine the extent of contribution by the respective cells. The enzyme level was estimated in cytosolic fraction, cell lysate and in conditioned media of these cells in rat and hamster. In both species the concentration of ADA was found to be markedly high in cytosolic fraction over to the cell lysate and the conditioned media in both TR and DC. Species-wise, it was higher in hamster. Cell-wise, the enzyme activity was significantly higher in TR than DC in rat but equal in hamster. In the conditioned medium, also, the enzyme activity was higher in TR in both species. The inference drawn from the results are: 1) the maximum enzyme activity in cytosolic fraction of TR and DC of both species clearly indicates equal involvement of the cells that constitute foeto-maternal unit, 2) the enhanced level of enzyme in TR and DC of hamster over to those of rat is possibly due to the higher proliferative activity in the cells of this species because of shorter gestation (16-17 days in hamster and 22-23 days in rats).     

Morphological and biochemical changes in old rat muscles: effect of increased use

Male Wistar rats were strength and swim trained during a substantial period of old age to determine the influence of aging and activity on the histochemical and metabolic characteristics of a predominantly slow (soleus) and a predominantly fast (plantaris) skeletal muscle. Strength training counteracted the age-related atrophy of the fibers and the age-induced changes in fiber-type distribution of both muscles. Swim training, on the other hand, was without any effect on these parameters. The activity of both mitochondrial and cytoplasmic enzymes became lower with aging in the soleus muscle, whereas only the activity of the cytoplasmic enzymes became lower in the plantaris. Strength training reduced the aerobic capacity of both muscles, whereas swim training had the opposite effect. Aging induced a lower glycogen concentration of the lateral gastrocnemius muscle. This was avoided by swim training. The phosphocreatine and adenosine 5'-triphosphate concentrations were unchanged with aging but became higher with strength training. The activity pattern, therefore, seems to have a considerable influence on the age-related modification of the histochemical and metabolic characteristics of skeletal muscles of the rat. The effect, however, is related to the recruitment pattern of the fiber populations and the form of activity.     

Binding site of the rat liver specific monoclonal antibody

A rat liver-specific antigen (RLSA) lost its binding ability to the corresponding monoclonal antibody after treatment with N-glycanase or sialidase, which suggested that the specific binding site might be in a portion of the sugar chain containing sialic acid. The specific antigen reacted with wheat germ agglutinin, lentil lectin, erythroagglutinating phytohemagglutinin and Ricinus communis agglutinin, but not with concanavalin A or peanut agglutinin. These results suggest that the specific antigen has asparagine-linked complex-type sugar chains which might be the binding sites of the monoclonal antibody.     

A specific gastrin receptor site in the rat stomach

A specific receptor for gastrin I has been demonstrated in the rat stomach fundus.Specific binding of ¹²⁵I-labelled gastrin I was localised to particles sedimenting between 250–20 000 × g. Saturation of binding sites occurred with a gastrin concentration of 10^?11 M in an assay system containing 0.6–1.7 mg/ml of homogenate protein. Gastrin binding was shown to be reversible, temperature- and pH-dependent, and susceptible to tryptic digestion. Electron microscopic and enzymatic studies showed the binding fraction to contain predominantly mitochondria. Preincubation of the homogenate with 10^?8 M cholecystokinin or secretin inhibited gastrin binding to a greater extent than an equimolar concentration of pentagastrin. Cimetidine, a histamine receptor antagonist, did not affect binding of gastrin to the receptor.     

Cytosolic phospholipase A2 in rat decidual cells: evidence for its role in decidualization

We investigated the existence and possible role of cytosolic phospholipase A2 (cPLA2) in rat decidualized uteri. PLA2 activity in the cytosol of a decidualized uterine horn, induced by intraluminal oil infusion, was significantly higher than that in contralateral intact horn. The activity was almost completely depressed by cPLA2 inhibitors including arachidonyl trifluoromethyl ketone (ATK). The immunoreactive signals for cPLA2 were intense in decidua and glandular epithelial cells. In vivo administration of ATK (0.1-100 microg) caused a dose-dependent inhibition of decidualization. These results show the presence of cPLA2 and its probable implication in decidualization in rat uterus.     

Apoptosis in activated rat pancreatic stellate cells

Proliferation and matrix synthesis by activated pancreatic stellate cells (PSC) participate in the development of chronic pancreatitis. Apoptosis of PSC may terminate this process but has not yet been studied in this particular cell type and was the aim of the present study. PSC were isolated from rat pancreas and characterized for expression of glial fibrillary acidic protein, alpha-smooth muscle actin, CD95, and tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) receptors. Apoptosis was determined by TdT-UTP nick end-labeling reaction, annexin V binding, and caspase-8 activation. Both CD95L and TRAIL induced apoptosis in PSC. The apoptotic response was minor in PSC cultured for 7 days but increased markedly thereafter. Sensitization of PSC with culture duration was accompanied by increased expression of CD95 and TRAIL receptor 2 and no alterations of Flip expression or protein kinase B phosphorylation but was paralleled by the appearance of a COOH-terminal cleavage product of receptor-interacting protein. PSC apoptosis was also induced by PK-11195, a ligand of the peripheral benzodiazepine receptor. PSC apoptosis may be important in terminating the wound-healing response after pancreas injury and exhibits features distinct from apoptosis induction in hepatic stellate cells.     

Electron microscopic study of the differentiation of decidual cells. I. The ultrastructure of large decidual cells in the antimesometrial portion of the rat decidua

The large decidual cells (LDC) of the antimesometrial part of decidua in rats of 7-9 days of gestation were studied by electron microscope. The decidual tissue has an epithelium-like pattern of organization. The apical surface of LDC is facing the pericapillar space making numerous villi. Lateral surfaces of these cells maintain close contact with each other by means of zona adhaerens, gap junctions, spot desmosomes and simple junctions. Accumulation of electron dense granules measuring from 0.05 to 0.3 mkm is seen in the apical parts of LDC. The Golgi complex and rough endoplasmic reticulum are much developed. The material of rough endoplasmic reticulum is denser than the cell matrix. Disperse chromatin is seen in the nucleus, whereas the granular component is dominanting in the nucleolus. It is concluded that the LDC may have a high metabolic activity, and that the secretion is a mode of fulfilling specific functions of LDC.     

A specific gastrin receptor site in the rat stomach.

A specific receptor for gastrin I has been demonstrated in the rat stomach fundus. Specific binding of 125I-labelled gastrin I was localised to particles sedimenting between 250--20 000 X g. Saturation of binding sites occurred with a gastrin concentration of 10(-11) M in an assay system containing 0.6--1.7 mg/ml of homogenate protein. Gastrin binding was shown to be reversible, temperature- and pH-dependent, and susceptible to tryptic digestion. Electron microscopic and enzymatic studies showed the binding fraction to contain predominantly mitochondria. Preincubation of the homogenate with 10(-8) M cholecystokinin or secretin inhibited gastrin binding to a greater extent than an equimolar concentration of pentagastrin. Cimetidine, a histamine receptor antagonist, did not affect binding of gastrin to the receptor.     

Prostaglandin and thromboxane production by rat decidual microsomes

The preparation of a microsomal fraction from the decidual tissue of pregnant rat uteri is described. Incubation of such microsomes with a mixture of radiolabelled and cold arachidonic acid (51 micrometer) plus cofactors resulted in a 30% substrate conversion. Products were resolved into four peaks (A, B, C and D) by thin-layer chromatography. Combined gas-liquid chromatography-mass spectrometry and further thin-layer chromatography identified the products as PGF2 alpha (A); thromboxane B2 (B); a mixture of 6-OXO PGF1 alpha and PGE2 (C); PGD2 and PGE2 (D). PGE2 was the major product.