Kwon Jeongwoo, Park Shuha, Seong Min-Jung, Choi Inchul, Kim Nam-Hyung
Department of Animal Sciences, Chungbuk National University, Naesudong-ro, Seowon-gu, Cheongju-si, Chungcheongbuk-do, 28864, Republic of Korea.
Department of Animal and Dairy Sciences, College of Agriculture and Life Sciences, Chungnam National University, Korea, 34134, Republic of Korea.
Reprod Fertil Dev. 2019 Jan;31(2):412-419. doi: 10.1071/RD18098.
Cytoplasmic polyadenylation element binding protein (CPEB) is an RNA-binding protein that promotes elongation of poly(A) tails and regulates mRNA translation. CPEB depletion in mammary epithelium is known to disrupt tight-junction (TJ) assembly via mislocalisation of tight junction protein 1 (TJP1), but the role of CPEB in the biological functions associated with TJs has not yet been studied. The objective of this study was to investigate the roles of CPEB2 during porcine parthenote development. CPEB2 was detected in both the nuclei and apical cytoplasm at the 4- and 8-cell stages and was localised to cell-cell contact after the initiation of the morula stage. Its depletion led to retarded blastocyst formation caused by impaired TJ assembly. Moreover, transcription of TJ-associated genes, including TJP1, Coxsackie virus and adenovirus receptor (CXADR) and occludin (OCLN), was not affected, but the corresponding proteins were not properly localised at the apical cell membrane in morulae, suggesting that CPEB2 confers mRNA stability or determines subcellular localisation for translation. Remarkably reduced relative levels of TJP1 transcripts bearing the 3'-untranslated region were noted, indicating that CPEB2 mediates TJP1 mRNA stability. In conclusion, our findings demonstrate that because of its regulation of TJP1, CPEB2 is required for TJ assembly during porcine blastocyst development.
细胞质聚腺苷酸化元件结合蛋白(CPEB)是一种RNA结合蛋白,可促进多聚腺苷酸(poly(A))尾巴的延长并调节mRNA翻译。已知乳腺上皮细胞中CPEB的缺失会通过紧密连接蛋白1(TJP1)的错误定位破坏紧密连接(TJ)组装,但CPEB在与TJ相关的生物学功能中的作用尚未得到研究。本研究的目的是探讨CPEB2在猪孤雌胚发育过程中的作用。在4细胞和8细胞阶段,CPEB2在细胞核和顶端细胞质中均有检测到,在桑葚胚阶段开始后定位于细胞间接触部位。其缺失导致TJ组装受损,从而导致囊胚形成延迟。此外,TJ相关基因的转录,包括TJP1、柯萨奇病毒和腺病毒受体(CXADR)以及闭合蛋白(OCLN),并未受到影响,但相应蛋白在桑葚胚中未正确定位于顶端细胞膜,这表明CPEB2赋予mRNA稳定性或决定翻译的亚细胞定位。值得注意 的是,携带3'-非翻译区的TJP1转录本的相对水平显著降低,表明CPEB2介导TJP1 mRNA的稳定性。总之,我们的研究结果表明,由于其对TJP1的调节作用,CPEB2是猪囊胚发育过程中TJ组装所必需的。