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

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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.


N-glycosylated ENPP3 in human endometrium.Deglycosylation of uterine fluid showed a shift of band from 165 kD to a single band at 110 kD, meaning that ENPP3 in the uterine fluid is present in its glycosylated form. The glycosylated (lanes 2–7) form of ENPP3 showed a band at 165 kD and the deglycosylated (lanes 8–13) form had a single band at 110 kD.
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f6: N-glycosylated ENPP3 in human endometrium.Deglycosylation of uterine fluid showed a shift of band from 165 kD to a single band at 110 kD, meaning that ENPP3 in the uterine fluid is present in its glycosylated form. The glycosylated (lanes 2–7) form of ENPP3 showed a band at 165 kD and the deglycosylated (lanes 8–13) form had a single band at 110 kD.

Mentions: The predicted molecular weight of ENPP3 is around 100 kd, but Western blot analysis showed a band at 165 kd, leading us to further investigate its glycosylation. We tested uterine fluid and endometrial tissue lysates after digesting with Peptide-N-Glycosidase F (PNGase F), which cleaves the glycoaminidase link between asparagine and N-acetylglucosamines. We observed a shift from 165 kD to 110 kD in the deglycosylated samples, confirming that ENPP3 is N-Glycosylated in the uterine fluid and endometria of healthy fertile women (Fig. 6).


Compartmentalized gene expression profiling of receptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form
N-glycosylated ENPP3 in human endometrium.Deglycosylation of uterine fluid showed a shift of band from 165 kD to a single band at 110 kD, meaning that ENPP3 in the uterine fluid is present in its glycosylated form. The glycosylated (lanes 2–7) form of ENPP3 showed a band at 165 kD and the deglycosylated (lanes 8–13) form had a single band at 110 kD.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036034&req=5

f6: N-glycosylated ENPP3 in human endometrium.Deglycosylation of uterine fluid showed a shift of band from 165 kD to a single band at 110 kD, meaning that ENPP3 in the uterine fluid is present in its glycosylated form. The glycosylated (lanes 2–7) form of ENPP3 showed a band at 165 kD and the deglycosylated (lanes 8–13) form had a single band at 110 kD.
Mentions: The predicted molecular weight of ENPP3 is around 100 kd, but Western blot analysis showed a band at 165 kd, leading us to further investigate its glycosylation. We tested uterine fluid and endometrial tissue lysates after digesting with Peptide-N-Glycosidase F (PNGase F), which cleaves the glycoaminidase link between asparagine and N-acetylglucosamines. We observed a shift from 165 kD to 110 kD in the deglycosylated samples, confirming that ENPP3 is N-Glycosylated in the uterine fluid and endometria of healthy fertile women (Fig. 6).

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.