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  1. Article ; Online: Porosomes in uterine epithelial cells: Ultrastructural identification and characterization during early pregnancy.

    Kalam, Sadaf N / Dowland, Samson / Lindsay, Laura / Murphy, Christopher R

    Journal of morphology

    2022  Volume 283, Issue 11, Page(s) 1381–1389

    Abstract: Porosomes are plasma membrane structures in secretory cells that allow transient docking and/or partial fusion of vesicles during which they release their content then disengage. This is referred to as "kiss and run" exocytosis. During early pregnancy, ... ...

    Abstract Porosomes are plasma membrane structures in secretory cells that allow transient docking and/or partial fusion of vesicles during which they release their content then disengage. This is referred to as "kiss and run" exocytosis. During early pregnancy, at the time of receptivity, there is a high level of vesicle activity in uterine epithelial cells (UECs). One of the secretory pathways for these vesicles could be via porosomes, which have yet to be identified in UECs. This study identified porosomes in the apical plasma membrane of UECs for the first time. These structures were present on days 1, 5.5, and 6 of early pregnancy, where they likely facilitate partial secretion via "kiss and run" exocytosis. The porosomes were measured and quantified on days 1, 5.5, and 6, which showed there are significantly more porosomes on day 5.5 (receptive) compared to day 1 (nonreceptive) of pregnancy. This increase in porosome numbers may reflect major morphological and molecular changes in the apical plasma membrane at this time such as increased cholesterol and soluble NSF attachment protein receptor proteins, as these are structural and functional components of the porosome complex assembly. Porosomes were observed in both resting (inactive) and dilated (active) states on days 1, 5.5, and 6 of early pregnancy. Porosomes on day 5.5 are significantly more active than on day 1 as demonstrated by the dilation of their base diameter. Further two-way ANOVA analysis of base diameter in resting and dilated states found a significant increase in porosome activity in day 5.5 compared to day 1. This study therefore indicates an increase in the number and activity of porosomes at the time of uterine receptivity in the rat, revealing a mechanism by which the UECs modify the uterine luminal environment at this time.
    MeSH term(s) Pregnancy ; Female ; Animals ; Rats ; Exocytosis ; Cell Membrane/metabolism ; Epithelial Cells ; Cholesterol/metabolism ; SNARE Proteins/metabolism
    Chemical Substances Cholesterol (97C5T2UQ7J) ; SNARE Proteins
    Language English
    Publishing date 2022-10-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 3084-3
    ISSN 1097-4687 ; 0022-2887 ; 0362-2525
    ISSN (online) 1097-4687
    ISSN 0022-2887 ; 0362-2525
    DOI 10.1002/jmor.21504
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Preschoolers Optimize the Timing of Their Conversational Turns Through Flexible Coordination of Language Comprehension and Production.

    Lindsay, Laura / Gambi, Chiara / Rabagliati, Hugh

    Psychological science

    2019  Volume 30, Issue 4, Page(s) 504–515

    Abstract: Conversation is the natural setting for language learning and use, and a key property of conversation is the smooth taking of turns. In adult conversations, delays between turns are minimal (typically 200 ms or less) because listeners display a striking ... ...

    Abstract Conversation is the natural setting for language learning and use, and a key property of conversation is the smooth taking of turns. In adult conversations, delays between turns are minimal (typically 200 ms or less) because listeners display a striking ability to predict what their partner will say, and they formulate a response before their partner's turn ends. Here, we tested how this ability to coordinate comprehension and production develops in preschool children. In an interactive paradigm, 106 children (ages 3-5 years) and 48 adults responded to questions that varied in predictability but were controlled for linguistic complexity. Using a novel distributional approach to data analysis, we found that when children can predict a question's ending, they leave shorter gaps before responding, suggesting that they can optimize the timing of their conversational turns like adults do. In line with a recent ethological theory of turn taking, this early competency helps explain how conversational contexts support language development.
    MeSH term(s) Adult ; Auditory Perception ; Child Language ; Child, Preschool ; Communication ; Comprehension ; Female ; Humans ; Interpersonal Relations ; Male
    Language English
    Publishing date 2019-02-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2022256-7
    ISSN 1467-9280 ; 0956-7976
    ISSN (online) 1467-9280
    ISSN 0956-7976
    DOI 10.1177/0956797618822802
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Membrane trafficking directed by VAMP2 and syntaxin 3 in uterine epithelial cells.

    Kalam, Sadaf N / Cole, Louise / Lindsay, Laura / Murphy, Christopher R

    Reproduction (Cambridge, England)

    2020  Volume 160, Issue 4, Page(s) 533–546

    Abstract: Luminal uterine epithelial cells (UEC) have a surge in vesicular activity during early uterine receptivity. It has been predicted these vesicles exit the UEC via exocytosis resulting in secretion and membrane trafficking. The present study investigated ... ...

    Abstract Luminal uterine epithelial cells (UEC) have a surge in vesicular activity during early uterine receptivity. It has been predicted these vesicles exit the UEC via exocytosis resulting in secretion and membrane trafficking. The present study investigated the changes in SNARE proteins VAMP2 (v-SNARE) and syntaxin 3 (t-SNARE) localisation and abundance in UECs during early pregnancy in the rat. We found VAMP2 and syntaxin 3 are significantly higher on day 5.5 compared to day 1 of pregnancy. On day 5.5, VAMP2 is perinuclear and syntaxin 3 is concentrated in the apical cytoplasm compared to a cytoplasmic localisation on day 1. This change in localisation and abundance show VAMP2 and syntaxin 3 are involved in vesicular movement and membrane trafficking in UECs during early pregnancy. This study also investigated the influence of cytoskeletal disruption of microtubules and actin filaments on VAMP2 and syntaxin 3 in UECs grown in vitro, since microtubules and actin influence vesicle trafficking. As expected, this study found disruption to microtubules with colchicine and actin with cytochalasin D impacted VAMP2 and syntaxin 3 localisation. These results suggest VAMP2 and syntaxin 3 are involved in the timely trafficking of vesicular membranes to the apical surface in UECs during early pregnancy, as are of microtubules and actin.
    MeSH term(s) Actins/metabolism ; Animals ; Cell Membrane/metabolism ; Cell Movement ; Cytoskeleton/metabolism ; Epithelial Cells/cytology ; Epithelial Cells/metabolism ; Exocytosis ; Female ; Pregnancy ; Protein Transport ; Qa-SNARE Proteins/metabolism ; Rats ; Rats, Wistar ; Uterus/cytology ; Uterus/metabolism ; Vesicle-Associated Membrane Protein 2/metabolism
    Chemical Substances Actins ; Qa-SNARE Proteins ; Vamp2 protein, rat ; Vesicle-Associated Membrane Protein 2
    Language English
    Publishing date 2020-07-21
    Publishing country England
    Document type Journal Article
    ZDB-ID 2034501-X
    ISSN 1741-7899 ; 1470-1626 ; 1476-3990
    ISSN (online) 1741-7899
    ISSN 1470-1626 ; 1476-3990
    DOI 10.1530/REP-20-0233
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Rab13 and Desmosome Redistribution in Uterine Epithelial Cells During Early Pregnancy.

    Lindsay, Laura A / Nasir, Reeja F / Dowland, Samson N / Madawala, Romanthi J / Murphy, Christopher R

    Reproductive sciences (Thousand Oaks, Calif.)

    2021  Volume 28, Issue 7, Page(s) 1981–1988

    Abstract: The luminal uterine epithelial cells are the first point of contact with the implanting blastocyst. Dramatic changes occur in the structure and function of these cells at the time of receptivity including changes in the lateral junctional complex. While ... ...

    Abstract The luminal uterine epithelial cells are the first point of contact with the implanting blastocyst. Dramatic changes occur in the structure and function of these cells at the time of receptivity including changes in the lateral junctional complex. While these morphological changes are important for uterine receptivity, currently there is no known mechanism of regulation of the lateral junctional complexes. Rab13, a member of the Rab (Ras-related in the brain) family of GTPases has a critical role in endosomal trafficking to the lateral plasma membrane and is involved in modulation of the tight junction in several cell types. The aim of this study is to investigate the role of Rab13 in changes to the lateral junctional complex at the time of receptivity. Immunofluorescence microscopy demonstrated no association between Rab13 and ZO-1 (a tight junction protein) or Rab13 and E-cadherin (an integral component of adherens junctions). Co-localisation was demonstrated between Rab 13 and desmoglein-2 at the time of fertilization and also at receptivity suggesting involvement of Rab13 in relocalisation of desmoglein-2 and formation of giant desmosomes in the apical part of the lateral plasma membrane at the time of uterine receptivity. We suggest that despite the loss of the adherens junction at the time of receptivity, the presently reported redistribution of desmosomes regulated by Rab13 allows the uterine epithelium to maintain structural integrity.
    MeSH term(s) Animals ; Cadherins/metabolism ; Desmosomes/metabolism ; Epithelial Cells/metabolism ; Female ; Pregnancy ; Rats ; Rats, Wistar ; Uterus/metabolism ; rab GTP-Binding Proteins/metabolism
    Chemical Substances Cadherins ; rab GTP-Binding Proteins (EC 3.6.5.2)
    Language English
    Publishing date 2021-02-01
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2276411-2
    ISSN 1933-7205 ; 1933-7191
    ISSN (online) 1933-7205
    ISSN 1933-7191
    DOI 10.1007/s43032-021-00478-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Change in distribution of cytoskeleton-associated proteins, lasp-1 and palladin, during uterine receptivity in the rat endometrium.

    Nicholson, Leigh / Lindsay, Laura / Murphy, Christopher R

    Reproduction, fertility, and development

    2018  Volume 30, Issue 11, Page(s) 1482–1490

    Abstract: The epithelium of the uterine lumen is the first point of contact with the blastocyst before implantation. To facilitate pregnancy, these uterine epithelial cells (UECs) undergo morphological changes specific to the receptive uterus. These changes ... ...

    Abstract The epithelium of the uterine lumen is the first point of contact with the blastocyst before implantation. To facilitate pregnancy, these uterine epithelial cells (UECs) undergo morphological changes specific to the receptive uterus. These changes include basal, lateral and apical alterations in the plasma membrane of UECs. This study looked at the cytoskeletal and focal adhesion-associated proteins, lasp-1 and palladin, in the uterus during early pregnancy in the rat. Two palladin isoforms, 140 kDa and 90 kDa, were analysed, with the migration-associated 140-kDa isoform increasing significantly at the time of implantation when compared with the time of fertilisation. Lasp-1 was similarly increased at this time, whilst also being located predominantly apically and laterally in the UECs, suggesting a role in the initial contact between the UECs and the blastocyst. This is the first study to investigate palladin and lasp-1 in the uterine luminal epithelium and suggests an importance for these cytoskeletal proteins in the morphological changes the UECs undergo for pregnancy to occur.
    MeSH term(s) Animals ; Cytoskeletal Proteins/genetics ; Cytoskeletal Proteins/metabolism ; Cytoskeleton/metabolism ; Embryo Implantation/physiology ; Endometrium/metabolism ; Epithelial Cells/metabolism ; Female ; Fertilization/physiology ; Microfilament Proteins/genetics ; Microfilament Proteins/metabolism ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Phosphoproteins/genetics ; Phosphoproteins/metabolism ; Pregnancy ; Rats
    Chemical Substances Cytoskeletal Proteins ; LOC103693936 ; Lasp1 protein, rat ; Microfilament Proteins ; Nerve Tissue Proteins ; Phosphoproteins
    Language English
    Publishing date 2018-05-05
    Publishing country Australia
    Document type Journal Article
    ZDB-ID 1019913-5
    ISSN 1448-5990 ; 1031-3613
    ISSN (online) 1448-5990
    ISSN 1031-3613
    DOI 10.1071/RD17530
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Altered immune environment in peritoneal endometriotic lesions: relationship to lesion appearance.

    Ersoy, Burcu / Hey-Cunningham, Alison J / Lindsay, Laura / Varol, Nesrin / Markham, Robert

    F&S science

    2020  Volume 2, Issue 2, Page(s) 207–218

    Abstract: Objective: To study the localization of and quantify different immune cell populations in red, black, and white peritoneal endometriotic lesions and compare immune cell densities between lesions and the surrounding tissue.: Design: Cross-sectional ... ...

    Abstract Objective: To study the localization of and quantify different immune cell populations in red, black, and white peritoneal endometriotic lesions and compare immune cell densities between lesions and the surrounding tissue.
    Design: Cross-sectional study.
    Setting: Teaching hospital, university research laboratory.
    Patient(s): Participants undergoing laparoscopic excision of endometriosis were recruited from gynecological operating theaters at Royal Prince Alfred Hospital, Sydney (n = 28).
    Intervention(s): Immunohistochemical staining for and quantification of dendritic cells (mature and immature), T cells (effector, cytotoxic, and regulatory), B cells, and macrophages in endometriotic peritoneal lesions and the surrounding tissue.
    Main outcome measure(s): Immune cell densities and aggregates were quantified.
    Result(s): Red and black lesions are significantly more likely to be surrounded by immune cell aggregates than white lesions (P=.036). In the tissue surrounding the peritoneal endometriotic lesions, there was a consistent pattern of greater and more variable density of immune cell populations for red lesions than black or white lesions and a range of significant positive correlations between densities of different immune populations (all P≤.004; not observed within the lesion stroma).
    Conclusion(s): There is a greater presence of immune cells in the tissue surrounding earlier/red and black lesions than older scarred white lesions, particularly in the form of immune cell aggregates, indicating an immunologic response in close proximity to the adjacent lesion. The relationship between densities of immune populations in the tissue surrounding the lesions suggests complementary recruitment and local interactions between cells. Categorizing immune cell populations in proximity to peritoneal endometriotic lesions may improve the understanding of lesion persistence and transition to older white appearances. Early (red) peritoneal endometriotic lesions are surrounded by a greater density of immune cells, including immune aggregates, than later (black or white) lesions. These immune cells may support lesion persistence.
    MeSH term(s) Cross-Sectional Studies ; Endometriosis/surgery ; Female ; Humans ; Laparoscopy ; Peritoneal Diseases/pathology ; Staining and Labeling
    Language English
    Publishing date 2020-11-20
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2666-335X
    ISSN (online) 2666-335X
    DOI 10.1016/j.xfss.2020.11.002
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  7. Article: Ovarian hyperstimulation affects fluid transporters in the uterus: a potential mechanism in uterine receptivity.

    Lindsay, Laura A / Murphy, Christopher R

    Reproduction, fertility, and development

    2014  Volume 26, Issue 7, Page(s) 982–990

    Abstract: Controlled ovarian hyperstimulation is commonly used in fertility treatment. Evidence suggests that this could alter the endometrial environment and influence implantation rate. However, the mechanisms underlying this disruption are unknown. A recently ... ...

    Abstract Controlled ovarian hyperstimulation is commonly used in fertility treatment. Evidence suggests that this could alter the endometrial environment and influence implantation rate. However, the mechanisms underlying this disruption are unknown. A recently developed rat ovarian hyperstimulation (OH) model found alterations in the localisation and expression of several molecules associated with implantation, as well as an increase in luminal fluid at the time of implantation. The present study investigated the effects of OH in rats on the expression of fluid-transporting molecules aquaporin 5 (AQP5) and claudin 4. The expression of these proteins was investigated in uterine luminal epithelial cells of rats undergoing OH and compared with normal pregnancy. There was a significant increase in AQP5 protein in OH rats at the time of implantation, along with a loss of the mesometrial staining gradient, which is thought to contribute to implantation position. At the same time, there was a significant decrease in claudin 4 protein. These results suggest that OH in rats causes a dysregulation in uterine fluid dynamics through modifications to fluid-transporting molecules, resulting in an unfavourable implantation environment for the blastocyst.
    MeSH term(s) Animals ; Aquaporin 5/analysis ; Claudin-4/analysis ; Cytoplasm/chemistry ; Embryo Implantation/drug effects ; Embryo Implantation/physiology ; Epithelial Cells/chemistry ; Female ; Gonadotropins, Equine/administration & dosage ; Ovulation Induction/adverse effects ; Ovulation Induction/methods ; Rats ; Rats, Wistar ; Tight Junctions/chemistry ; Uterus/chemistry ; Uterus/physiology ; Uterus/ultrastructure
    Chemical Substances Aquaporin 5 ; Claudin-4 ; Gonadotropins, Equine
    Language English
    Publishing date 2014-08
    Publishing country Australia
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1019913-5
    ISSN 1448-5990 ; 1031-3613
    ISSN (online) 1448-5990
    ISSN 1031-3613
    DOI 10.1071/RD12396
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Microtubules are reorganised and fragmented for uterine receptivity.

    Kalam, Sadaf N / Dowland, Samson / Lindsay, Laura / Murphy, Christopher R

    Cell and tissue research

    2018  Volume 374, Issue 3, Page(s) 667–677

    Abstract: For the development of uterine receptivity, many morphological and molecular changes occur in the apical surface of luminal uterine epithelial cells (UECs) including an increase in vesicular activity. Vesicular movements for exocytosis and endocytosis ... ...

    Abstract For the development of uterine receptivity, many morphological and molecular changes occur in the apical surface of luminal uterine epithelial cells (UECs) including an increase in vesicular activity. Vesicular movements for exocytosis and endocytosis are dependent on microtubules; however, changes in microtubules in UECs during early pregnancy have received little attention. β-tubulin, one of the main component of microtubules, is distributed throughout the cytoplasm of UECs on day 1 (non-receptive) of pregnancy in the rat. On day 5.5, β-tubulin is concentrated above the nuclei and by day 6 (receptive), β-tubulin is concentrated in a band-like fashion above the nucleus. Western blot analysis of isolated UECs found two bands (50 and 34 kDa) for β-tubulin in UECs during early pregnancy. The intensity of the 34 kDa band was significantly higher on day 6 compared to day 1. The increase in the 34 kDa band may be due to higher proteolytic activity associated with microtubule polymerisation during the receptive state. Transmission electron microscopy showed fragmented microtubules at the time of receptivity in UECs. This is the first study to show that microtubules are reorganised during uterine receptivity. This re-organisation likely facilitates vesicular movement and promotes the reorganisation of the apical plasma membrane for uterine receptivity.
    MeSH term(s) Animals ; Cell Separation ; Epithelial Cells/metabolism ; Female ; Microtubules/metabolism ; Microtubules/ultrastructure ; Pregnancy ; Rats, Wistar ; Tubulin/metabolism ; Uterus/cytology ; Uterus/metabolism
    Chemical Substances Tubulin
    Language English
    Publishing date 2018-07-20
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 125067-x
    ISSN 1432-0878 ; 0302-766X
    ISSN (online) 1432-0878
    ISSN 0302-766X
    DOI 10.1007/s00441-018-2887-x
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  9. Article ; Online: α-Parvin and β-parvin in the rat uterus during decidualisation and uterine receptivity.

    Nicholson, Leigh / Madawala, Romanthi / Lindsay, Laura / Murphy, Christopher R

    Histochemistry and cell biology

    2018  Volume 151, Issue 5, Page(s) 395–406

    Abstract: During early pregnancy, the uterine luminal epithelial cells (UECs) and endometrial stromal cells (ESCs) undergo morphological changes to enable blastocyst implantation. The present study investigates, for the first time, the cytoskeletal-associated ... ...

    Abstract During early pregnancy, the uterine luminal epithelial cells (UECs) and endometrial stromal cells (ESCs) undergo morphological changes to enable blastocyst implantation. The present study investigates, for the first time, the cytoskeletal-associated proteins and α-actinin superfamily members, α-parvin and β-parvin, during early pregnancy in the rat uterus. These two PARVA proteins are involved in cell adhesion, morphological changes and regulation of other cytoskeletal proteins, through binding with proteins such as actin and integrin-linked kinase. α-parvin is present in UECs at fertilisation and significantly decreases by the time of implantation. β-parvin acts in opposition; significantly increasing in both UECs and ESCs at the time of implantation, suggesting a role in the process of decidualisation. Additionally, the presence of a serine-8 residue-phosphorylated α-parvin, which is associated with cell morphology changes, was found in the nuclear region of both UECs and ESCs during implantation and decidualisation. We also show that the presence of both β-parvin and phosphorylated α-parvin in ESCs is dependent on decidualisation occurring. This study demonstrates that the changing balance and localisation of the two PARVA proteins are dependent on the time of uterine receptivity, suggesting a co-dependent role in the cytoskeletal re-organisation crucial to the changing conditions necessary for implantation and decidualisation.
    MeSH term(s) Actinin/metabolism ; Animals ; Female ; Male ; Pregnancy ; Rats ; Rats, Wistar ; Uterus/cytology ; Uterus/metabolism
    Chemical Substances Actinin (11003-00-2)
    Language English
    Publishing date 2018-12-05
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 1222930-1
    ISSN 1432-119X ; 0301-5564 ; 0948-6143
    ISSN (online) 1432-119X
    ISSN 0301-5564 ; 0948-6143
    DOI 10.1007/s00418-018-1761-y
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Uterine focal adhesions are retained at implantation after rat ovarian hyperstimulation.

    Lindsay, Laura A / Dowland, Samson N / Murphy, Christopher R

    Reproduction (Cambridge, England)

    2016  Volume 152, Issue 6, Page(s) 753–763

    Abstract: Controlled ovarian hyperstimulation is an essential component of IVF techniques to ensure proliferation and development of multiple ovarian follicles, but the effects of these hormones on the endometrium are largely unknown. During normal pregnancy in ... ...

    Abstract Controlled ovarian hyperstimulation is an essential component of IVF techniques to ensure proliferation and development of multiple ovarian follicles, but the effects of these hormones on the endometrium are largely unknown. During normal pregnancy in rats, there are significant changes in the basal plasma membrane of uterine epithelial cells (UECs) at the time of receptivity, including loss of focal adhesions. This enables the UECs to be removed from the implantation chamber surrounding the blastocyst, thus allowing invasion into the underlying stroma. This study investigated the influence of ovarian hyperstimulation (OH) on the basal plasma membrane of UECs during early pregnancy in the rat. Immunofluorescence results demonstrate the presence of paxillin, talin, integrin β1 and phosphorylated FAK (Y397FAK) in the basal portion of UECs at the time of implantation in OH pregnancy. TEM analysis demonstrated a flattened basal lamina and the presence of focal adhesions on the basal surface at this time in OH pregnancy. Significantly low full-length paxillin, high paxillin δ and integrin β1 were seen at the time of implantation in OH compared with those in normal pregnancy. The increase in paxillin δ suggests that these cells are less mobile, whereas the increase in integrin β1 and Y397FAK suggests the retention of a stable FA complex. Taken together with the increase in morphological focal adhesions, this represents a cell type that is stable and less easily removed for blastocyst implantation. This may be one mechanism explaining lower implantation rates after fresh embryo transfers compared with frozen cycles.
    MeSH term(s) Animals ; Cell Membrane/metabolism ; Embryo Implantation ; Female ; Focal Adhesions/metabolism ; Focal Adhesions/pathology ; Ovarian Hyperstimulation Syndrome/physiopathology ; Rats ; Rats, Wistar ; Uterus/metabolism ; Uterus/pathology
    Language English
    Publishing date 2016-12
    Publishing country England
    Document type Journal Article
    ZDB-ID 2034501-X
    ISSN 1741-7899 ; 1470-1626 ; 1476-3990
    ISSN (online) 1741-7899
    ISSN 1470-1626 ; 1476-3990
    DOI 10.1530/REP-16-0331
    Database MEDical Literature Analysis and Retrieval System OnLINE

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