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Compartmentalized gene expression profiling of receptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form

View Article: PubMed Central - PubMed

ABSTRACT

The complexity of endometrial receptivity at the molecular level needs to be explored in detail to improve the management of infertility. Here, differential expression of transcriptomes in receptive endometrial glands and stroma revealed Ectonucleotide Pyrophosphatase/Phosphodiesterase 3 (ENPP3) as a progesterone regulated factor and confirmed by various methods, both at mRNA and protein level. The involvement of ENPP3 in embryo attachment was tested in an in vitro model for human embryo implantation. Interestingly, there was high expression of ENPP3 mRNA in stroma but not protein. Presence of N-glycosylated ENPP3 in receptive phase uterine fluid in women confirms its regulation by progesterone and makes it possible to use in a non-invasive test of endometrial receptivity.

No MeSH data available.


Expression of ENPP3 in endometrium and uterine fluid.Western blot analysis of ENPP3 in receptive phase endometrial tissue lysates (A,B) and uterine fluid (C,D) by Wes showed good expression levels of ENPP3 in control samples (lanes 2–7). Antiprogestin treatment (A,C): lanes 8–13) showed no detectable levels of ENPP3, both in endometrial tissue and uterine fluid, confirming the regulation of ENPP3 protein by progesterone. (B,D) show semiquantitative analysis of immunodetectable ENPP3 by Wes, expressed in log2 AUC (area under the curve). Lane 1: protein molecular weight marker.
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f5: Expression of ENPP3 in endometrium and uterine fluid.Western blot analysis of ENPP3 in receptive phase endometrial tissue lysates (A,B) and uterine fluid (C,D) by Wes showed good expression levels of ENPP3 in control samples (lanes 2–7). Antiprogestin treatment (A,C): lanes 8–13) showed no detectable levels of ENPP3, both in endometrial tissue and uterine fluid, confirming the regulation of ENPP3 protein by progesterone. (B,D) show semiquantitative analysis of immunodetectable ENPP3 by Wes, expressed in log2 AUC (area under the curve). Lane 1: protein molecular weight marker.

Mentions: The uterine lavages from fertile women and the same women treated with a single dose of 200 mg of mifepristone on LH + 2 were collected on LH + 6/7 and tested for ENPP3 protein expression by the western blotting technique. A strong band was observed around 165 kd indicating glycosylated ENPP3, and showed a significant downregulation in P-inhibited group (AUC - control 17.85: mifepristone treatment: 9.47; p = 0.002) with a similar pattern to that of tissue ENPP3 (AUC -control 18.98: mifepristone treatment 11.94; p = 0.002). The expression of ENPP3 in endometrial tissue lysate was more abundant than in uterine fluid (Fig. 5).


Compartmentalized gene expression profiling of receptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form
Expression of ENPP3 in endometrium and uterine fluid.Western blot analysis of ENPP3 in receptive phase endometrial tissue lysates (A,B) and uterine fluid (C,D) by Wes showed good expression levels of ENPP3 in control samples (lanes 2–7). Antiprogestin treatment (A,C): lanes 8–13) showed no detectable levels of ENPP3, both in endometrial tissue and uterine fluid, confirming the regulation of ENPP3 protein by progesterone. (B,D) show semiquantitative analysis of immunodetectable ENPP3 by Wes, expressed in log2 AUC (area under the curve). Lane 1: protein molecular weight marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC5036034&req=5

f5: Expression of ENPP3 in endometrium and uterine fluid.Western blot analysis of ENPP3 in receptive phase endometrial tissue lysates (A,B) and uterine fluid (C,D) by Wes showed good expression levels of ENPP3 in control samples (lanes 2–7). Antiprogestin treatment (A,C): lanes 8–13) showed no detectable levels of ENPP3, both in endometrial tissue and uterine fluid, confirming the regulation of ENPP3 protein by progesterone. (B,D) show semiquantitative analysis of immunodetectable ENPP3 by Wes, expressed in log2 AUC (area under the curve). Lane 1: protein molecular weight marker.
Mentions: The uterine lavages from fertile women and the same women treated with a single dose of 200 mg of mifepristone on LH + 2 were collected on LH + 6/7 and tested for ENPP3 protein expression by the western blotting technique. A strong band was observed around 165 kd indicating glycosylated ENPP3, and showed a significant downregulation in P-inhibited group (AUC - control 17.85: mifepristone treatment: 9.47; p = 0.002) with a similar pattern to that of tissue ENPP3 (AUC -control 18.98: mifepristone treatment 11.94; p = 0.002). The expression of ENPP3 in endometrial tissue lysate was more abundant than in uterine fluid (Fig. 5).

View Article: PubMed Central - PubMed

ABSTRACT

The complexity of endometrial receptivity at the molecular level needs to be explored in detail to improve the management of infertility. Here, differential expression of transcriptomes in receptive endometrial glands and stroma revealed Ectonucleotide Pyrophosphatase/Phosphodiesterase 3 (ENPP3) as a progesterone regulated factor and confirmed by various methods, both at mRNA and protein level. The involvement of ENPP3 in embryo attachment was tested in an in vitro model for human embryo implantation. Interestingly, there was high expression of ENPP3 mRNA in stroma but not protein. Presence of N-glycosylated ENPP3 in receptive phase uterine fluid in women confirms its regulation by progesterone and makes it possible to use in a non-invasive test of endometrial receptivity.

No MeSH data available.