Detection of the pan neuronal marker PGP9.5 by immuno-histochemistry and quantitative PCR in eutopic endometrium from women with and without endometriosis

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PGP9.5 mRNA was elevated in the proliferative phase of endometriosis patients with pain, and PGP9.5 protein was present in endometriosis patients' endometrium but not controls, with no correlation between gene and protein levels.

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The study assessed neuroendocrine and endometriosis-associated product PGP9.5 at both mRNA and protein levels in eutopic endometrium from 29 women undergoing laparoscopy (16 with endometriosis, 13 controls), using qPCR and immuno-histochemistry on samples taken by curettage, and examined associations with pain symptoms and menstrual cycle phase. PGP9.5 mRNA did not differ overall between women with and without endometriosis, but was higher in women reporting pain (not statistically significant) and was significantly higher in the proliferative phase among endometriosis patients with pain and in the first half of the cycle in endometriosis patients versus controls. Immuno-histochemistry showed PGP9.5 immunoreactivity in 13/16 endometriosis patients and none in women without endometriosis, while nerve fibre density per mm² was similar in endometriosis patients regardless of pain status; the paper emphasizes lack of a direct gene–protein correspondence. This paper is centrally about endometriosis — specifically, detecting and comparing PGP9.5/neuronal marker expression and eutopic endometrial nerve fibres in women with vs without endometriosis.

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Abstract

PurposeTo assess endometrial gene as well as protein expression of neuroendocrine and supposedly endometriosis-associated product PGP9.5 and pain symptoms in women with endometriosis and controls undergoing laparoscopy, using molecular biological and immuno-histochemical approaches in the same patients.MethodsBiopsy of eutopic endometrium from 29 patients by sharp curettage, and preparation of paraffin blocks. Determination of PGP9.5 gene expression and protein abundance using qPCR and immuno-histochemistry.ResultsqPCR; The PGP9.5 mRNA expression level between women with (N = 16) and without (N = 13) endometriosis was not different, regardless of pain symptoms or menstrual cycle phase. PGP9.5 expression was higher in women who reported pain compared to those who did not; however, this association was not statistically significant. The expression of PGP9.5 mRNA was higher in women with endometriosis and pain during the proliferative than in the secretory phase (P = 0.03). Furthermore, in the first half of the cycle, the abundance of the PGP9.5 transcript was also significantly higher in endometriosis patients compared to those without (P = 0.03). Immuno-histochemistry; Thirteen of the 16 endometriosis patients showed positive PGP9.5 immuno-reactivity in the endometrium, whereas no such signal was observed in women without endometriosis. The absolute number of nerve fibres per mm(2) in women with endometriosis was similar, regardless of the pain symptoms.ConclusionsPGP9.5 mRNA expression is increased in the proliferative phase of endometriotic women with pain. The presence of nerve fibres was demonstrated by a PGP9.5 protein signal in immuno-histochemistry and restricted to patients with endometriosis. Based on these results, however, there did not appear to be a direct association between the gene expression and protein abundance in women with and without endometriosis or those that experienced pain.
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Abstract

Purpose To assess endometrial gene as well as protein expression of neuroendocrine and supposedly endometriosis-associated product PGP9.5 and pain symptoms in women with endometriosis and controls undergoing laparoscopy, using molecular biological and immuno-histochemical approaches in the same patients.

Methods

Biopsy of eutopic endometrium from 29 patients by sharp curettage, and preparation of paraffin blocks. Determination of PGP9.5 gene expression and protein abundance using qPCR and immuno-histochemistry.

Results

qPCR; The PGP9.5 mRNA expression level between women with (N = 16) and without (N = 13) endometriosis was not different, regardless of pain symptoms or menstrual cycle phase. PGP9.5 expression was higher in women who reported pain compared to those who did not; however, this association was not statistically significant. The expression of PGP9.5 mRNA was higher in women with endometriosis and pain during the proliferative than in the secretory phase (P = 0.03). Furthermore, in the first half of the cycle, the abundance of the PGP9.5 transcript was also significantly higher in endometriosis patients compared to those without (P = 0.03). Immuno-histochemistry; Thirteen of the 16 endometriosis patients showed positive PGP9.5 immuno-reactivity in the endometrium, whereas no such signal was observed in women without endometriosis. The absolute number of nerve fibres per mm2 in women with endometriosis was similar, regardless of the pain symptoms.

Conclusions

PGP9.5 mRNA expression is increased in the proliferative phase of endometriotic women with pain. The presence of nerve fibres was demonstrated by a PGP9.5 protein signal in immuno-histochemistry and restricted to patients with endometriosis. Based on these results, however, there did not appear to be a direct association between the gene expression and protein abundance in women with and without endometriosis or those that experienced pain. Similar content being viewed by others

References

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Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Zevallos, H.BV., McKinnon, B., Tokushige, N. et al. Detection of the pan neuronal marker PGP9.5 by immuno-histochemistry and quantitative PCR in eutopic endometrium from women with and without endometriosis. Arch Gynecol Obstet 291, 85–91 (2015). https://doi.org/10.1007/s00404-014-3379-1 Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s00404-014-3379-1

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Condition tags

endometriosis

MeSH descriptors

Endometriosis Endometrium Pain Ubiquitin Thiolesterase Biomarkers Biomarkers Biopsy Endometriosis Endometrium Female Humans Immunohistochemistry Laparoscopy Middle Aged Nerve Fibers Nerve Fibers Pain Polymerase Chain Reaction RNA, Messenger RNA, Messenger

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