Difference in expression between AQP1 and AQP5 in porcine endometrium and myometrium in response to steroid hormones, oxytocin, arachidonic acid, forskolin and cAMP during the mid-luteal phase of the estrous cycle and luteolysis

Abstract

BackgroundRecently, we demonstrated in vitro that AQP1 and AQP5 in the porcine uterus are regulated by steroid hormones (P4, E2), arachidonic acid (AA), forskolin (FSK) and cAMP during the estrous cycle. However, the potential of the porcine separated uterine tissues, the endometrium and myometrium, to express these AQPs remains unknown. Thus, in this study, the responses of AQP1 and AQP5 to P4, E2 oxytocin (OT), AA, FSK and cAMP in the porcine endometrium and myometrium were examined during the mid-luteal phase of the estrous cycle and luteolysis.MethodsReal-time PCR and western blot analysis.ResultsProgesterone up-regulated the expression of AQP1/AQP5 mRNAs and proteins in the endometrium and myometrium, especially during luteolysis. Similarly, E2 also stimulated the expression of both AQPs, but only in the endometrium. AA led to the upregulation of AQP1/AQP5 in the endometrium during luteolysis. In turn, OT increased the expression of AQP1/AQP5 mRNAs and proteins in the myometrium during mid-luteal phase. Moreover, a stimulatory effect of forskolin and cAMP on the expression of AQP1/AQP5 mRNAs and proteins in the endometrium and myometrium dominated during luteolysis, but during the mid-luteal phase their influence on the expression of these AQPs was differentiated depending on the type of tissue and the incubation duration.ConclusionsThese results seem to indicate that uterine tissues; endometrium and myometrium, exhibit their own AQP expression profiles in response to examined factors. Moreover, the responses of AQP1/AQP5 at mRNA and protein levels to the studied factors in the endometrium and myometrium are more pronounced during luteolysis. This suggests that the above effects of the studied factors are connected with morphological and physiological changes taking place in the pig uterus during the estrous cycle.

The In Vitro Effect of Steroid Hormones, Arachidonic Acid, and Kinases Inhibitors on Aquaporin 1, 2, 5, and 7 Gene Expression in the Porcine Uterine Luminal Epithelial Cells during the Estrous Cycle

“…Our experiments confirmed the AQP1 and AQP5 gene expression in the porcine uterine luminal epithelial cells [11,[25][26][27], a novel finding was that AQP2 and AQP7 are also expressed in these cells. In addition, in this study, we investigated the effects of the estradiol, progesterone, arachidonic acid, H89 (selective PKA inhibitor), and PD98059 (MAPK inhibitor) on the AQP1, AQP2, AQP5, and AQP7 gene expression in the uterine epithelial cells.…”

“…In the transcellular pathway, aquaporins are mainly responsible for a large amount of transport of water, which is driven by the osmotic gradient [6]. It has been already shown that several factors, including hormones, regulate uterine transepithelial water transport via modulating the expression level of AQPs [6,[25][26][27]40]. As shown in our results, treatment with steroid hormones (E 2 and P 4 ) resulted in a predominantly stimulatory effect on AQP1 and AQP2 mRNA expression.…”

“…In our recent study, we demonstrated the endometrial and myometrial expression of AQP1, 5, and 9 during different phases of the estrous cycle and pregnancy, which indicates that steroid hormones, arachidonic acid (AA), oxytocin (OT), forskolin (FSK), and cyclic adenosine monophosphate (cAMP) participate in controlling the distribution of AQP1 and 5 in the uterus [11,[25][26][27]. Thus, following our previous results, the aim of this study was to (1) examine the gene expression of AQP1, 2, 5, and 7 in the porcine uterine epithelial cells during the periovulatory period; (2) determine the effect of E 2 , P 4 , and AA on the expression of studied AQPs in these cells; (3) find out whether the examined factors affect the PKA and MAPK signaling pathways during the periovulatory period.…”

Pituitary Gonadotropins, Prolactin and Growth Hormone Differentially Regulate AQP1 Expression in the Porcine Ovarian Follicular Cells

“…They stimulate follicular cell proliferation and, together with FSH, initiate the formation of LH receptors in granulosa cells, mainly in the mural, progesterone and androgen production in theca interna cells [ 37 , 38 ]. A thorough discussion regarding the influence of steroid hormones on aquaporins in the female reproductive system has been already presented in our previous papers [ 39 , 40 , 41 ]. In turn, very interesting results on transgenic mice deficient in AQP proteins showed different reproductive phenomena in both males and females [ 42 , 43 , 44 , 45 ].…”

Seminal Plasma Modulates miRNA Expression by Sow Genital Tract Lining Explants

“…These findings suggest that these secreted miRNAs could also modulate the maternal intrauterine environment. To the best of our knowledge, this is the first study examining miRNA expression profiles in the culture media harvested after exposure of different segments of the sow genital tract (explants from uterus, UTJ and isthmus) to SP (from SRF, post-SRF and EE) for 16 h. Oviduct and uterine explants have been cultured from several mammalian species (including pigs), and widely used as a standardized model to evaluate in vitro female genital tract responses to different treatments, due to the fact that the epithelium is exposed, as in the in vivo situation [ 44 , 45 , 46 , 47 , 48 ]. Although this ex vivo model may not completely reflect the changes that could take place in vivo, it provides reliable and useful data that can contribute to a better understanding about the role played by SP on the secretion of miRNAs by female genital tract.…”