Diel de Amorim Mariana, Klein Claudia, Foster Robert, Dong Lynn, Lopez-Rodriguez Maria Fernanda, Card Claire
Department of Clinical Sciences, Cornell University, 930 Campus Rd, Ithaca, NY 14853, USA.
Department of Veterinary Clinical and Diagnostic Science, University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4, Canada.
Animals (Basel). 2022 Mar 22;12(7):799. doi: 10.3390/ani12070799.
Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is an enzyme that metabolizes oxytocin in serum and tissues. The presence of oxytocin/neurophysin I (OXT), oxytocin and LNPEP and their relationship to other genes is unknown in the equine conceptus. Our objective was to characterize gene expression of LNPEP and OXT on D8, 10, 12, 14, 15, 16 and 21 conceptuses in relationship to other genes. Immunohistochemistry, western blot and liquid chromatography with tandem mass spectrometry (LC-MS/MS) were used for identification of oxytocin and LNPEP in D15, 16 and 18 conceptuses. LNPEP was increased at D15 compared to D10, was immunolocalized in the equine trophectoderm and endoderm, and protein was confirmed by LC-MS/MS. Maximal abundance of OXT was at D21, and lowest on D12 and D14, but no protein was identified. OXTR abundance was highest on D14 and D21. LNPEP was correlated with PTGFR and PTGES on D12 and D14-D15, and high expression of PTGES, PTGS2 was found on D14, D15 and D21; PTGFR was found on D8 and D12-21. LNPEP may have a role in prostaglandin regulation and conceptus fixation by decreasing the availability of oxytocin. Further investigation on the role embryonic LNPEP during pregnancy is warranted.
亮氨酰和胱氨酰氨肽酶(LNPEP/催产素酶)是一种在血清和组织中代谢催产素的酶。在马的孕体中,催产素/神经垂体素I(OXT)、催产素和LNPEP的存在及其与其他基因的关系尚不清楚。我们的目的是表征孕体在第8、10、12、14、15、16和21天时LNPEP和OXT的基因表达,并研究其与其他基因的关系。采用免疫组织化学、蛋白质印迹法和液相色谱-串联质谱法(LC-MS/MS)对第15、16和18天孕体中的催产素和LNPEP进行鉴定。与第10天相比,第15天时LNPEP增加,其免疫定位在马的滋养外胚层和内胚层,并且通过LC-MS/MS证实了该蛋白的存在。OXT的丰度在第21天时最高,在第12天和第14天时最低,但未鉴定出蛋白质。OXTR的丰度在第14天和第21天时最高。在第12天以及第14 - 15天时,LNPEP与PTGFR和PTGES相关,在第14、15和21天时发现PTGES、PTGS2高表达;在第8天以及第12 - 21天时发现PTGFR。LNPEP可能通过降低催产素的可用性在前列腺素调节和孕体着床中发挥作用。有必要进一步研究胚胎LNPEP在妊娠期间的作用。