scholarly journals Redistribution of aquaporins 1 and 5 in the rat uterus is dependent on progesterone: a study with light and electron microscopy

Reproduction ◽  
2006 ◽  
Vol 131 (2) ◽  
pp. 369-378 ◽  
Author(s):  
Laura A Lindsay ◽  
Christopher R Murphy
Keyword(s):  
Protein Expression ◽  
Light And Electron Microscopy ◽  
Normal Pregnancy ◽  
Apical Plasma Membrane ◽  
Uterine Receptivity ◽  
Rat Uterus ◽  
Oestrogen Treatment ◽  
Aqp5 Expression ◽  
Circular Layer ◽  
Blastocyst Implantation

During early pregnancy in the rat there is a dramatic reduction in luminal fluid which is associated with uterine receptivity for blastocyst implantation. This study investigates the presence and distributional changes of several members of the aquaporin (AQP) family in the rat uterus in response to hormonal regime. An increase in apical AQP5 protein expression was found in response to progesterone alone or in combination with oestrogen, which is similar to that seen at the time of implantation. AQP1 was found in endothelial cells of the endometrium and in the inner circular layer of smooth muscle, with maximal protein expression seen after three doses of progesterone plus 8 hr of oestrogen treatment. These results, for the first time, show that the up-regulation of AQP5 in the apical plasma membrane of uterine epithelial cells and AQP1 in the inner circular layer of myometrium, is dependent on progesterone. Furthermore, unlike during normal pregnancy, there is no differential gradient of AQP5 expression between mesometrial and antimesometrial poles of the progesterone treated uterus. Hence it is suggested that the differential gradient of AQP5 is dependent on the presence of a blastocyst, in addition to the appropriate hormonal environment.

Changes in the fine structure of the apical plasma membrane of endometrial epithelial cells during implantation in the rat

1982 ◽  
Vol 55 (1) ◽  
pp. 1-12
Author(s):  
C.R. Murphy ◽  
J.G. Swift ◽  
T.M. Mukherjee ◽  
A.W. Rogers
Keyword(s):  
Plasma Membrane ◽  
Fine Structure ◽  
Epithelial Cells ◽  
Normal Pregnancy ◽  
Ovariectomized Rats ◽  
Apical Plasma Membrane ◽  
Embryonic Cells ◽  
Blastocyst Implantation ◽  
First Contact ◽  
Endometrial Epithelial Cells

In previous work we have shown that ovarian hormones, when injected into ovariectomized rats, alter the fine structure of the plasma membrane of endometrial epithelial cells. In this paper freeze-fractures have been used to study the apical plasma membrane of endometrial epithelial cells of rats during the period of blastocyst implantation of normal pregnancy. On day 1 of pregnancy there were 2354 +/− 114 intramembranous particles (IMPs) per micrometer2 of membrane. The particles were spherical and randomly distributed. On day 5 of pregnancy IMP density rose to 2899 +/− 289 per micrometer2 and some rod-shaped particles were also visible. By day 6 of pregnancy IMP density had risen to 4014 +/− 206 per micrometer2 and there were more rod-shaped IMPs than before. In addition, on day 6 IMPs were also present as rows of particles and some gap-junction-like arrays of particles were also seen. Our findings indicate that there are fine-structural alterations in the apical plasma membrane of endometrial epithelial cells, the site of first contact between maternal and embryonic cells, during the period of early pregnancy. The findings are discussed in the light of suggested mechanisms of blastocyst attachment to the uterine epithelium at implantation.

Actin crosslinking protein filamin A during early pregnancy in the rat uterus

2016 ◽  
Vol 28 (7) ◽  
pp. 960 ◽  
Author(s):  
Romanthi J. Madawala ◽  
Connie E. Poon ◽  
Samson N. Dowland ◽  
Christopher R. Murphy
Keyword(s):  
Actin Cytoskeleton ◽  
Early Pregnancy ◽  
Filamin A ◽  
Uterine Receptivity ◽  
Rat Uterus ◽  
Decidual Cells ◽  
Cell Layers ◽  
Important Regulator ◽  
Blastocyst Implantation ◽  
Major Transformation

During early pregnancy the endometrium undergoes a major transformation in order for it to become receptive to blastocyst implantation. The actin cytoskeleton and plasma membrane of luminal uterine epithelial cells (UECs) and the underlying stromal cells undergo dramatic remodelling to facilitate these changes. Filamin A (FLNA), a protein that crosslinks actin filaments and also mediates the anchorage of membrane proteins to the actin cytoskeleton, was investigated in the rat uterus at fertilisation (Day 1) and implantation (Day 6) to determine the role of FLNA in actin cytoskeletal remodelling of UECs and decidua during early pregnancy. Localisation of FLNA in UECs at the time of fertilisation was cytoplasmic, whilst at implantation it was distributed apically; its localisation is under the influence of progesterone. FLNA was also concentrated to the first two to three stromal cell layers at the time of fertilisation and shifted to the primary decidualisation zone at the time of implantation. This shift in localisation was found to be dependent on the decidualisation reaction. Protein abundance of the FLNA 280-kDa monomer and calpain-cleaved fragment (240 kDa) did not change during early pregnancy in UECs. Since major actin cytoskeletal remodelling occurs during early pregnancy in UECs and in decidual cells, the changing localisation of FLNA suggests that it may be an important regulator of cytoskeletal remodelling of these cells to allow uterine receptivity and decidualisation necessary for implantation in the rat.

scholarly journals Maternal Neutrophil Depletion Fails to Avert Systemic Lipopolysaccharide-Induced Early Pregnancy Defects in Mice

2021 ◽  
Vol 22 (15) ◽  
pp. 7932
Author(s):  
Sourav Panja ◽  
John T. Benjamin ◽  
Bibhash C. Paria
Keyword(s):  
Early Pregnancy ◽  
Normal Pregnancy ◽  
Mrna Levels ◽  
Interesting Approach ◽  
Blastocyst Implantation ◽  
Neutrophil Depletion ◽  
Underlying Mechanisms ◽  
Myd88 Signaling ◽  
Induced Defects ◽  
Dose Dependent

Maternal infection-induced early pregnancy complications arise from perturbation of the immune environment at the uterine early blastocyst implantation site (EBIS), yet the underlying mechanisms remain unclear. Here, we demonstrated in a mouse model that the progression of normal pregnancy from days 4 to 6 induced steady migration of leukocytes away from the uterine decidual stromal zone (DSZ) that surrounds the implanted blastocyst. Uterine macrophages were found to be CD206+ M2-polarized. While monocytes were nearly absent in the DSZ, DSZ cells were found to express monocyte marker protein Ly6C. Systemic endotoxic lipopolysaccharide (LPS) exposure on day 5 of pregnancy led to: (1) rapid (at 2 h) induction of neutrophil chemoattractants that promoted huge neutrophil infiltrations at the EBISs by 24 h; (2) rapid (at 2 h) elevation of mRNA levels of MyD88, but not Trif, modulated cytokines at the EBISs; and (3) dose-dependent EBIS defects by day 7 of pregnancy. Yet, elimination of maternal neutrophils using anti-Ly6G antibody prior to LPS exposure failed to avert LPS-induced EBIS defects allowing us to suggest that activation of Tlr4-MyD88 dependent inflammatory pathway is involved in LPS-induced defects at EBISs. Thus, blocking the activation of the Tlr4-MyD88 signaling pathway may be an interesting approach to prevent infection-induced pathology at EBISs.

PBA increases CFTR expression but at high doses inhibits Cl−secretion in Calu-3 airway epithelial cells

1999 ◽  
Vol 277 (4) ◽  
pp. L700-L708 ◽  
Author(s):  
Johannes Loffing ◽  
Bryan D. Moyer ◽  
Donna Reynolds ◽  
Bruce A. Stanton
Keyword(s):  
Protein Expression ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Airway Epithelial Cells ◽  
Cell Phenotype ◽  
Apical Plasma Membrane ◽  
Airway Epithelial ◽  
High Concentration ◽  
Cycle Enzyme ◽  
Cf Transmembrane Conductance Regulator

Sodium 4-phenylbutyrate (PBA), a short-chain fatty acid, has been approved to treat patients with urea cycle enzyme deficiencies and is being evaluated in the management of sickle cell disease, thalassemia, cancer, and cystic fibrosis (CF). Because relatively little is known about the effects of PBA on the expression and function of the wild-type CF transmembrane conductance regulator (wt CFTR), the goal of this study was to examine the effects of PBA and related compounds on wt CFTR-mediated Cl−secretion. To this end, we studied Calu-3 cells, a human airway cell line that expresses endogenous wt CFTR and has a serous cell phenotype. We report that chronic treatment of Calu-3 cells with a high concentration (5 mM) of PBA, sodium butyrate, or sodium valproate but not of sodium acetate reduced basal and 8-(4-chlorophenylthio)-cAMP-stimulated Cl−secretion. Paradoxically, PBA enhanced CFTR protein expression 6- to 10-fold and increased the intensity of CFTR staining in the apical plasma membrane. PBA also increased protein expression of Na+-K+-ATPase. PBA reduced CFTR Cl−currents across the apical membrane but had no effect on Na+-K+-ATPase activity in the basolateral membrane. Thus a high concentration of PBA (5 mM) reduces Cl−secretion by inhibiting CFTR Cl−currents across the apical membrane. In contrast, lower therapeutic concentrations of PBA (0.05–2 mM) had no effect on cAMP-stimulated Cl−secretion across Calu-3 cells. We conclude that PBA concentrations in the therapeutic range are unlikely to have a negative effect on Cl−secretion. However, concentrations >5 mM might reduce transepithelial Cl−secretion by serous cells in submucosal glands in individuals expressing wt CFTR.

scholarly journals Effect ofAtractylodes macrocephalaon Hypertonic Stress-Induced Water Channel Protein Expression in Renal Collecting Duct Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Yong Pyo Lee ◽  
Yun Jung Lee ◽  
So Min Lee ◽  
Jung Joo Yoon ◽  
Hye Yoom Kim ◽  
...  
Keyword(s):  
Protein Expression ◽  
Collecting Duct ◽  
Water Channel ◽  
Immunoblot Analysis ◽  
The Body ◽  
Apical Plasma Membrane ◽  
Diuretic Effect ◽  
Hypertonic Stress ◽  
Abnormal Accumulation ◽  
Atractylodes Macrocephala

Edema is a symptom that results from the abnormal accumulation of fluid in the body. The cause of edema is related to the level of aquaporin (AQP)2 protein expression, which regulates the reabsorption of water in the kidney. Edema is caused by overexpression of the AQP2 protein when the concentration of Na+in the blood increases. The rhizome ofAtractylodes macrocephalahas been used in traditional oriental medicine as a diuretic drug; however, the mechanism responsible for the diuretic effect of the aqueous extract fromA. macrocephalarhizomes (AAMs) has not yet been identified. We examined the effect of the AAM on the regulation of water channels in the mouse inner medullary collecting duct (mIMCD)-3 cells under hypertonic stress. Pretreatment of AAM attenuates a hypertonicity-induced increase in AQP2 expression as well as the trafficking of AQP2 to the apical plasma membrane. Tonicity-responsive enhancer binding protein (TonEBP) is a transcription factor known to play a central role in cellular homeostasis by regulating the expression of some proteins, including AQP2. Western immunoblot analysis demonstrated that the protein and mRNA expression levels of TonEBP also decrease after AAM treatment. These results suggest that the AAM has a diuretic effect by suppressing water reabsorption via the downregulation of the TonEBP-AQP2 signaling pathway.

PLASMA AND PITUITARY LUTEINIZING HORMONE CONCENTRATIONS AND PERIPHERAL PLASMA OESTRADIOL CONCENTRATION DURING EARLY PREGNANCY AND AFTER THE ADMINISTRATION OF PROGESTATIONAL STEROIDS IN THE RAT

1972 ◽  
Vol 53 (1) ◽  
pp. 31-35 ◽  
Author(s):  
K. BROWN-GRANT ◽  
C. S. CORKER ◽  
F. NAFTOLIN
Keyword(s):  
Luteinizing Hormone ◽  
Marked Decrease ◽  
Single Injection ◽  
Normal Pregnancy ◽  
Oestrous Cycle ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
Peripheral Plasma ◽  
Blastocyst Implantation ◽  
Plasma Oestradiol

SUMMARY Plasma luteinizing hormone (LH) concentrations were already lower on Day 2 of pregnancy than at the same time after the preceding ovulation in the non-pregnant rat, and fell progressively up to Day 16 of pregnancy. No evidence was obtained of any increase at the time when the ovulatory surge of LH would have occurred if the animal had not become pregnant. Pituitary LH concentration was lower in mated rats on the morning of Day 0 of pregnancy than in unmated controls on the morning of the day of oestrus. Subsequently it increased slowly to reach a level higher than at any stage of the oestrous cycle by Day 8 of pregnancy and remained high until at least Day 16 of pregnancy. Peripheral plasma oestradiol concentration increased late on Day 2 of pregnancy and was still raised on Day 4 but was never more than about one fourth of the peak concentration seen on the morning of prooestrus during the oestrous cycle. There were similar changes in plasma LH and oestradiol concentrations in the 48 h after a single injection of 2·5mg progesterone on the morning of the day of dioestrus, a procedure that delays ovulation by 1 or 2 days. Administration of a synthetic progestational compound (medroxyprogesterone acetate) to pregnant rats delayed blastocyst implantation and the delay was associated with a marked decrease in peripheral plasma LH to levels below those of normal pregnancy.

310. CAVEOLIN 1 AND 2 IN THE UTERUS DURING EARLY PREGNANCY

2010 ◽  
Vol 22 (9) ◽  
pp. 110
Author(s):  
R. J. Madawala ◽  
C. R. Murphy
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Early Pregnancy ◽  
Basolateral Membrane ◽  
Membrane Curvature ◽  
Major Protein ◽  
Uterine Epithelial Cells ◽  
Caveolin 1 ◽  
Blastocyst Implantation

Rat uterine epithelial cells undergo many changes during early pregnancy in order to become receptive to blastocyst implantation. These changes include basolateral folding and the presence of vesicles of various sizes which are at their greatest number during the pre-implantation period. The present study investigated the possible role that caveolin 1 and 2 plays in this remodelling specifically days 1, 3, 6, 7, and 9 of pregnancy. Caveolin is a major protein in omega shaped invaginations of the plasma membrane called caveolae that are considered to be specialised plasma membrane subdomains. Caveolae are rich in cholesterol, glycosphingolipids, and GPI anchored proteins and are involved in endocytosis and membrane curvature. Immunofluorescence microscopy has shown caveolin 1 and 2 on day 1 of pregnancy are localised to the cytoplasm of luminal uterine epithelial cells, and by day 6 of pregnancy (the time of implantation), it concentrates basally. By day 9 of pregnancy, expression of both caveolin 1 and 2 in luminal uterine epithelia is cytoplasmic as seen on day 1 of pregnancy. A corresponding increase in protein expression of caveolin 1 on day 6 of pregnancy in luminal uterine epithelia was observed. Interestingly however, caveolin 2 protein expression decreases at the time of implantation as found by western blot analysis. Both caveolin 1 and 2 were localised to blood vessels within the endometrium and myometrium and also the muscle of the myometrium in all days of pregnancy studied. In addition, both caveolin 1 and 2 were absent from glandular epithelium, which is interesting considering that they do not undergo the plasma membrane transformation. The localisation and expression of caveolin 1 and 2 in rat luminal uterine epithelium at the time of implantation suggest possible roles in trafficking of cholesterol and/or various proteins for either degradation or relocation. Caveolins may contribute to the morphology of the basolateral membrane seen on day 6 of pregnancy. All of which may play an important role during successful blastocyst implantation.

scholarly journals Progesterone supplementation extends uterine receptivity for blastocyst implantation in mice

Reproduction ◽  
2007 ◽  
Vol 133 (2) ◽  
pp. 487-493 ◽  
Author(s):  
Haengseok Song ◽  
Kyuyong Han ◽  
Hyunjung Lim
Keyword(s):  
Cell Proliferation ◽  
Embryo Transfer ◽  
Thymidine Incorporation ◽  
Expression Patterns ◽  
Physiological Changes ◽  
Uterine Receptivity ◽  
Blastocyst Implantation ◽  
Progesterone Supplementation

We previously showed that blastocyst can initiate implantation beyond the normal ‘window’ of uterine receptivity on day 5 of pregnancy and pseudopregnancy (PSP) in mice. In this study, we investigated whether uterine receptivity for blastocyst implantation can be further extended on day 6 of PSP and the role of progesterone (P4) on this event. Embryo transfers, experimentally induced decidualization,in situhybridization and [3H]thymidine incorporation were performed. Blastocysts initiate attachment reaction within 48 h when transferred on day 5, but not on day 6 of PSP. Likewise, decidualization reaction occurred on days 4 and 5 of PSP, but completely failed on day 6. However, P4supplementation partially retains uterine receptivity for blastocyst implantation and decidualization on day 6 of PSP. In addition, certain indicators of uterine receptivity, such as cell proliferation profile and expression patterns of implantation-related genes were similarly observed on days 4 and 5 of PSP, but not on day 6. Consistent with embryo transfer and decidualization, exogenous administration of P4partially restores these indicators on day 6 of PSP. We concluded that critical physiological changes occur between days 4 and 5 of PSP, leading to uterine non-receptivity on day 6, but P4is able to extend the uterine receptivity through day 6.

scholarly journals Extended uterine receptivity for blastocyst implantation and full-term fetal development in mice with vitrified-warmed ovarian tissue autotransplantation

2012 ◽  
Vol 11 (3) ◽  
pp. 123-128 ◽  
Author(s):  
Hiromichi Matsumoto ◽  
Kenji Ezoe ◽  
Akinori Mitsui ◽  
Emiko Fukui ◽  
Masanori Ochi ◽  
...  
Keyword(s):  
Fetal Development ◽  
Ovarian Tissue ◽  
Full Term ◽  
Uterine Receptivity ◽  
Blastocyst Implantation
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