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Generation and Characterization of Rat Uterus Organoids from Rat Endometrial Epithelial Stem Cells
Published on: August 2, 2024
Estrogen-dependent up-regulation of TRPA1 and TRPV1 receptor proteins in the rat endometrium
Krisztina Pohóczky1, József Kun2, Bálint Szalontai3
1Department of Pharmacology and PharmacotherapyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryJanos Szentagothai Research CentreUniversity of Pécs, Ifjúság Street 20, H-7624 Pécs, HungaryDepartments of PathologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryDepartment of PhysiologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryDepartment of Pathophysiology and GerontologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryMTA-PTE NAP B Chronic Pain Research GroupHungary, Szigeti Street 12, H-7624 Pécs, HungaryInstitute of Animal ScienceCentre for Agricultural and Applied Economic Sciences, University of Debrecen, PO Box 36, H-4015 Debrecen, Hungary Department of Pharmacology and PharmacotherapyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryJanos Szentagothai Research CentreUniversity of Pécs, Ifjúság Street 20, H-7624 Pécs, HungaryDepartments of PathologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryDepartment of PhysiologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryDepartment of Pathophysiology and GerontologyUniversity of Pécs Medical School, Szigeti Street 12, H-7624 Pécs, HungaryMTA-PTE NAP B Chronic Pain Research GroupHungary, Szigeti Street 12, H-7624 Pécs, HungaryInstitute of Animal ScienceCentre for Agricultural and Applied Economic Sciences, University of Debrecen, PO Box 36, H-4015 Debrecen, Hungary.
Abstract:
Transient receptor potential ankyrin 1 (TRPA1) and vanilloid 1 (TRPV1) receptors expressed predominantly in sensory nerves are activated by inflammatory stimuli and mediate inflammation and pain. Although they have been shown in the human endometrium, their regulation and function are unknown. Therefore, we investigated their estrogen- and progesterone-dependent alterations in the rat endometrium in comparison with the estrogen-regulated inflammatory cytokine macrophage migration inhibitory factor (MIF). Four-week-old (sexually immature) and four-month-old (sexually mature) female rats were treated with the non-selective estrogen receptor (ER) agonist diethylstilboestrol (DES), progesterone and their combination, or ovariectomized. RT-PCR and immunohistochemistry were performed to determine mRNA and protein expression levels respectively. Channel function was investigated with ratiometric [Ca(2+)]i measurement in cultured primary rat endometrial cells. Both TRP receptors and MIF were detected in the endometrium at mRNA and protein levels, and their localizations were similar. Immunostaining was observed in the immature epithelium, while stromal, glandular and epithelial positivity were observed in adults. Functionally active TRP receptor proteins were shown in endometrial cells by activation-induced calcium influx. In adults, Trpa1 and Trpv1 mRNA levels were significantly up-regulated after DES treatment. TRPA1 increased after every treatment, but TRPV1 remained unchanged following the combined treatment and ovariectomy. In immature rats, DES treatment resulted in increased mRNA expression of both channels and elevated TRPV1 immunopositivity. MIF expression changed in parallel with TRPA1/TRPV1 in most cases. DES up-regulated Trpa1, Trpv1 and Mif mRNA levels in endometrial cell cultures, but 17β-oestradiol having ERα-selective potency increased only the expression of Trpv1. We provide the first evidence for TRPA1/TRPV1 expression and their estrogen-induced up-regulation in the rat endometrium in correlation with the MIF.
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