Lenis Yasser Y, Johnson Gregory A, Wang Xiaoqiu, Tang Wendy W, Dunlap Kathrin A, Satterfield M Carey, Wu Guoyao, Hansen Thomas R, Bazer Fuller W
1Department of Animal Science, Texas A&M University, College Station, TX 77843-2471 USA.
2Center for Animal Biotechnology and Genomics, Texas A&M University, College Station, TX 77843 USA.
J Anim Sci Biotechnol. 2018 Jan 24;9:10. doi: 10.1186/s40104-017-0225-x. eCollection 2018.
Polyamines stimulate DNA transcription and mRNA translation for protein synthesis in trophectoderm cells, as well as proliferation and migration of cells; therefore, they are essential for development and survival of conceptuses (embryo/fetus and placenta). The ovine conceptus produces polyamines via classical and non-classical pathways. In the classical pathway, arginine (Arg) is transformed into ornithine, which is then decarboxylated by ornithine decarboxylase (ODC1) to produce putrescine which is the substrate for the production of spermidine and spermine. In the non-classical pathway, Arg is converted to agmatine (Agm) by arginine decarboxylase (ADC), and Agm is converted to putrescine by agmatinase (AGMAT).
Morpholino antisense oligonucleotides (MAOs) were designed and synthesized to inhibit translational initiation of the mRNAs for ODC1 and ADC, in ovine conceptuses.
The morphologies of MAO control, MAO-ODC1, and MAO-ADC conceptuses were normal. Double knockdown of and (MAO-ODC1:ADC) resulted in two phenotypes of conceptuses; 33% of conceptuses appeared to be morphologically and functionally normal (phenotype a) and 67% of the conceptuses presented an abnormal morphology and functionality (phenotype b). Furthermore, MAO-ODC1:ADC (a) conceptuses had greater tissue concentrations of Agm, putrescine, and spermidine than MAO control conceptuses, while MAO-ODC1:ADC (b) conceptuses only had greater tissue concentrations of Agm . Uterine flushes from ewes with MAO-ODC1:ADC (a) had greater amounts of arginine, aspartate, tyrosine, citrulline, lysine, phenylalanine, isoleucine, leucine, and glutamine, while uterine flushes of ewes with MAO-ODC1:ADC (b) conceptuses had lower amount of putrescine, spermidine, spermine, alanine, aspartate, glutamine, tyrosine, phenylalanine, isoleucine, leucine, and lysine.
The double-knockdown of translation of and mRNAs was most detrimental to conceptus development and their production of interferon tau (IFNT). Agm, polyamines, amino acids, and adequate secretion of IFNT are critical for establishment and maintenance of pregnancy during the peri-implantation period of gestation in sheep.
多胺可刺激滋养外胚层细胞中DNA转录和用于蛋白质合成的mRNA翻译，以及细胞的增殖和迁移；因此，它们对于孕体（胚胎/胎儿和胎盘）的发育和存活至关重要。绵羊孕体通过经典途径和非经典途径产生多胺。在经典途径中，精氨酸（Arg）转化为鸟氨酸，然后鸟氨酸脱羧酶（ODC1）将其脱羧生成腐胺，腐胺是生成亚精胺和精胺的底物。在非经典途径中，精氨酸脱羧酶（ADC）将精氨酸转化为胍丁胺（Agm），胍丁胺酶（AGMAT）将胍丁胺转化为腐胺。
设计并合成吗啉代反义寡核苷酸（MAO），以抑制绵羊孕体中ODC1和ADC的mRNA的翻译起始。
MAO对照、MAO-ODC1和MAO-ADC孕体的形态正常。ODC1和ADC的双重敲低（MAO-ODC1:ADC）导致孕体出现两种表型；33%的孕体在形态和功能上似乎正常（表型a），67%的孕体呈现异常的形态和功能（表型b）。此外，MAO-ODC1:ADC（a）孕体的组织中胍丁胺、腐胺和亚精胺的浓度高于MAO对照孕体，而MAO-ODC1:ADC（b）孕体的组织中仅胍丁胺浓度较高。来自怀有MAO-ODC1:ADC（a）孕体的母羊子宫冲洗液中精氨酸、天冬氨酸、酪氨酸、瓜氨酸、赖氨酸、苯丙氨酸、异亮氨酸、亮氨酸和谷氨酰胺的含量较高，而怀有MAO-ODC1:ADC（b）孕体的母羊子宫冲洗液中腐胺、亚精胺、精胺、丙氨酸、天冬氨酸、谷氨酰胺、酪氨酸丶苯丙氨酸、异亮氨酸、亮氨酸和赖氨酸的含量较低。
ODC1和ADC的mRNA翻译的双重敲低对孕体发育及其干扰素τ（IFNT）的产生最为不利。胍丁胺、多胺、氨基酸以及IFNT的充分分泌对于绵羊妊娠植入前期妊娠的建立和维持至关重要。
