Expression and regulation of prostaglandin transporters, ATP-binding cassette, subfamily C, member 1 and 9, and solute carrier organic anion transporter family, member 2A1 and 5A1 in the uterine endometrium during the estrous cycle and pregnancy in pigs원문보기
Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic an...
Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.
Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.
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제안 방법
Data from real-time RT-PCR for ABCC1, ABCC9, SLCO4C1, and SLCO5A1 expression during the estrous cycle and pregnancy were subjected to least squares analysis of variance using the general linear models procedures of SAS (Cary, NC, USA). As sources of variation, the model included day, pregnancy status (cyclic or pregnant, D12 and D15 post-estrus), and their interactions, to evaluate steady-state levels of ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs, and effects of treatment and animal, to evaluate effects of steroid hormones on ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs. Preplanned orthogonal contrasts (control vs E2; control vs P4; E2 vs E2+P4; E2+P4 vs E2+P4+ICI; and E2+P4 vs E2+P4+RU) were used to test for effects of treatments in the explant cultures.
Explant tissues were then collected and total RNA or protein was extracted for real-time RT-PCR and immunoblot analysis, respectively, to determine the expression levels of ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs. These experiments were conducted using endometrium on D12 of the estrous cycle, and treatments were performed in triplicate using tissues obtained from three gilts.
대상 데이터
This study was supported by the Next Generation BioGreen 21 Program (#PJ01119103), Rural Development Administration, Republic of Korea.
데이터처리
Data from real-time RT-PCR for ABCC1, ABCC9, SLCO4C1, and SLCO5A1 expression during the estrous cycle and pregnancy were subjected to least squares analysis of variance using the general linear models procedures of SAS (Cary, NC, USA). As sources of variation, the model included day, pregnancy status (cyclic or pregnant, D12 and D15 post-estrus), and their interactions, to evaluate steady-state levels of ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs, and effects of treatment and animal, to evaluate effects of steroid hormones on ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs.
Preplanned orthogonal contrasts (control vs E2; control vs P4; E2 vs E2+P4; E2+P4 vs E2+P4+ICI; and E2+P4 vs E2+P4+RU) were used to test for effects of treatments in the explant cultures. Data from real-time RT-PCR performed to assess the effect of the day of pregnancy (D12, D15, D30, D60, D90, and D114) in the endometrium, effect of the day of pregnancy (D30, D60, D90, and D114) in chorioallantoic tissue, on the expression of ABCC1, ABCC9, SLCO4C1, and SLCO5A1, and data from IL1B dose-response studies were analyzed by least squares regression analysis. Data are presented as least squares means with SEM.
이론/모형
The levels of expression of ABCC1, ABCC9, SLCO4C1, and SLCO5A1 genes in endometrial and chorioallantoic tissues were analyzed by real-time RT-PCR using the Applied Biosystems StepOnePlus System (Applied Biosystems, Foster City, CA, USA) and SYBR Green method. cDNAs were synthesized from 4 μg each of the total RNA isolated from different uterine endometrial tissues.
성능/효과
Results are reported as expression relative to that detected on D12 of the estrous cycle or that detected in control explant tissues after normalization of the transcript amount to the endogenous ribosomal protein L7 (RPL7) control by the 2–ΔΔCT method as previously described [19].
However, to our knowledge, expression of SLCO4C1 and SLCO5A1 in placental tissues has not been reported so far. Results of this study showed that conceptus tissues during early pregnancy and chorioallantoic tissues during mid to late pregnancy expressed ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs, and levels of ABCC1 and ABCC9 mRNAs in chorioallantoic membrane changed during mid to late pregnancy. Interestingly, SLCO5A1 mRNA, which was expressed at the highest level in the uterine endometrium on D12 of pregnancy, was detectable in conceptus tissues on D12 of pregnancy, but not on D15 of pregnancy.
Results of this study showed that levels of ABCC1 and SLCO2A1 mRNA were significantly higher on D12 of pregnancy than the estrous cycle and levels of ABCC9 and SLCO4C1 mRNA were higher during pregnancy than during the estrous cycle. In pigs, conceptus produces estrogen and IL1B2 during the implantation period [4].
In summary, this study showed that i) ABCC1, ABCC9, SLCO4C1, and SLCO5A1 genes are expressed in pregnancy stage and cell type specific manner in the uterine endometrium, ii) their expression during the implantation period is regulated by E2 or IL1B in the endometrial tissues, and iii) conceptuses during early pregnancy and chorioallantoic tissues during mid to late pregnancy also express ABCC1, ABCC9, SLCO4C1, and SLCO5A1. Our findings suggest that ABCC1, ABCC9, SLCO4C1, and SLCO5A1 play an important role in PG transport at the maternal-fetal interface for the establishment and maintenance of pregnancy in pigs.
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