University of Veterinary Medicine, Department of Obstetrics and Reproduction, István str 2., H-1078 Budapest, Hungary.
University of Veterinary Medicine, Department of Obstetrics and Reproduction, István str 2., H-1078 Budapest, Hungary.
Reprod Biol. 2020 Mar;20(1):9-13. doi: 10.1016/j.repbio.2020.01.005. Epub 2020 Jan 18.
Pituitary adenylate cyclase activating polypeptide (PACAP) was originally isolated as a hypothalamic neuropeptide stimulating adenylate cyclase activity. Besides its neuroprotective effects, numerous data proved its role in reproductive processes. However, there are limited data on its role in preimplantation embryo development and implantation. Our aim was to analyse the mRNA expression of Adcyap1 (coding region of PACAP) and Hbegf [coding region of HB-EGF (Heparin-binding EGF-like growth factor)] in embryos and pregnant uterus to investigate the possible correlation between them. Eight-week-old BDF1 mice were superovulated and subsequently mated overnight or left in their cage after hCG treatment. Day4 embryos were flushed from mated females. After morphological analysis, Adcyap1 and Hbegf gene expression of embryos and uterine tissues was assessed with qPCR. Our results showed significantly higher Adcyap1 and Hbegf mRNA levels in females producing embryos compared to non-mated ones. Robust elevation of Adcyap1 and slight elevation of Hbegf were detected in females with blastocyst embryos compared with non-blastocysts. We found low rate of Hbegf mRNA expression in uncompacted embryos, whereas morulae and blastocysts expressed high amounts of Hbegf. However, we did not find detectable Adcyap1 mRNA in embryos. Strong correlation was found between uterine tissue and embryonic Hbegf levels, slight correlation between uterine Adcyap1 and Hbegf levels. Uterine tissue Adcyap1 and embryonic Hbegf showed no correlation. In summary, our present data show, for the first time, the correlation between PACAP and HB-EGF mRNA expression suggesting that PACAP might play a role during the peri-implantation period of early mouse embryo development.
垂体腺苷酸环化酶激活肽 (PACAP) 最初是作为一种刺激腺苷酸环化酶活性的下丘脑神经肽被分离出来的。除了其神经保护作用外，大量数据还证明了它在生殖过程中的作用。然而，关于其在胚胎着床前发育和着床中的作用的数据有限。我们的目的是分析 Adcyap1（PACAP 编码区）和 Hbegf[编码区的 HB-EGF（肝素结合 EGF 样生长因子）]在胚胎和妊娠子宫中的 mRNA 表达，以研究它们之间的可能相关性。8 周龄的 BDF1 小鼠被超排卵，随后在 hCG 处理后与雄鼠过夜交配或留在笼子里。从交配后的雌性小鼠中冲洗出第 4 天的胚胎。在形态分析后，使用 qPCR 评估胚胎和子宫组织中 Adcyap1 和 Hbegf 基因的表达。我们的结果表明，与未交配的雌性相比，产生胚胎的雌性的 Adcyap1 和 Hbegf mRNA 水平显著升高。与非囊胚相比，具有囊胚胚胎的雌性的 Adcyap1 和 Hbegf 水平明显升高。我们发现未压缩胚胎的 Hbegf mRNA 表达率较低，而桑葚胚和囊胚表达大量的 Hbegf。然而，我们在胚胎中没有检测到可检测的 Adcyap1 mRNA。在子宫组织和胚胎 Hbegf 水平之间发现了很强的相关性，在子宫组织 Adcyap1 和 Hbegf 水平之间发现了轻微的相关性。子宫组织 Adcyap1 和胚胎 Hbegf 之间没有相关性。总之，我们目前的数据首次显示了 PACAP 和 HB-EGF mRNA 表达之间的相关性，表明 PACAP 可能在早期小鼠胚胎发育的着床期发挥作用。
