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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.

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Regulation of ENPP3 by progesterone.The expression of ENPP3 was observed very specifically in the apical border of glands in P dominant mid-luteal phase as studied by immunohistochemistry (A,C,D) in women without mifepristone treatment (control). Downregulation of ENPP3 was observed in endometrial glands with the suppression of progesterone action by mifepristone treatment (B,C). In endometrium of women without mifepristone treatment, high level of ENPP3 was observed during P upregulated secretory phase, on comparing with proliferative phase of the menstrual cycle. Scale bar indicates 50 μm.
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f4: Regulation of ENPP3 by progesterone.The expression of ENPP3 was observed very specifically in the apical border of glands in P dominant mid-luteal phase as studied by immunohistochemistry (A,C,D) in women without mifepristone treatment (control). Downregulation of ENPP3 was observed in endometrial glands with the suppression of progesterone action by mifepristone treatment (B,C). In endometrium of women without mifepristone treatment, high level of ENPP3 was observed during P upregulated secretory phase, on comparing with proliferative phase of the menstrual cycle. Scale bar indicates 50 μm.

Mentions: Based on the literature review, microarray expression levels, and availability of antibodies, we studied the protein expression of stanniocalcin 1 (STC1); Cathepsin C (CTSC); Secrteoglobin family 2A member2 (SCGB2A2); SWI/SNF related matrix associated actin-dependent regulator of chromatin subfamily a member 1 (SMARCA1); high-mobility group nucleosome binding domain 5 (HMGN5); and B-cell CLL/lymphoma 11A (zinc finger protein) (BCL11A). Immunodetection of ENPP3 was observed in all the tissues from healthy fertile women and differed significantly between the groups. The findings were in line with the microarray results (Fig. 4A–C).


Compartmentalized gene expression profiling of receptive endometrium reveals progesterone regulated ENPP3 is differentially expressed and secreted in glycosylated form
Regulation of ENPP3 by progesterone.The expression of ENPP3 was observed very specifically in the apical border of glands in P dominant mid-luteal phase as studied by immunohistochemistry (A,C,D) in women without mifepristone treatment (control). Downregulation of ENPP3 was observed in endometrial glands with the suppression of progesterone action by mifepristone treatment (B,C). In endometrium of women without mifepristone treatment, high level of ENPP3 was observed during P upregulated secretory phase, on comparing with proliferative phase of the menstrual cycle. Scale bar indicates 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Regulation of ENPP3 by progesterone.The expression of ENPP3 was observed very specifically in the apical border of glands in P dominant mid-luteal phase as studied by immunohistochemistry (A,C,D) in women without mifepristone treatment (control). Downregulation of ENPP3 was observed in endometrial glands with the suppression of progesterone action by mifepristone treatment (B,C). In endometrium of women without mifepristone treatment, high level of ENPP3 was observed during P upregulated secretory phase, on comparing with proliferative phase of the menstrual cycle. Scale bar indicates 50 μm.
Mentions: Based on the literature review, microarray expression levels, and availability of antibodies, we studied the protein expression of stanniocalcin 1 (STC1); Cathepsin C (CTSC); Secrteoglobin family 2A member2 (SCGB2A2); SWI/SNF related matrix associated actin-dependent regulator of chromatin subfamily a member 1 (SMARCA1); high-mobility group nucleosome binding domain 5 (HMGN5); and B-cell CLL/lymphoma 11A (zinc finger protein) (BCL11A). Immunodetection of ENPP3 was observed in all the tissues from healthy fertile women and differed significantly between the groups. The findings were in line with the microarray results (Fig. 4A–C).

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.


Related in: MedlinePlus