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首页> 外文期刊>Nucleic Acids Research >Coordinate control of cell cycle regulatory genes in zebrafish development tested by cyclin D1 knockdown with morpholino phosphorodiamidates and hydroxyprolyl-phosphono peptide nucleic acids.
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Coordinate control of cell cycle regulatory genes in zebrafish development tested by cyclin D1 knockdown with morpholino phosphorodiamidates and hydroxyprolyl-phosphono peptide nucleic acids.

机译：斑马鱼发育中细胞周期调控基因的协调控制，通过用吗啉代磷酸二酰胺酯和羟脯氨酰基膦酰基肽核酸进行的细胞周期蛋白D1敲除测试。

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During early zebrafish (Danio rerio) development zygotic transcription does not begin until the mid-blastula transition (MBT) 3 h after fertilization. MBT demarcates transition from synchronous short cell cycles of S and M phases exclusively to full cycles encompassing G1 and G2 phases. Transcriptional profiling and RT-PCR analyses during these phases enabled us to determine that this shift corresponds to decreased transcript levels of S/M phase cell cycle control genes (e.g. ccna2, ccnb1, ccnb2 and ccne) and increased transcript levels of ccnd1, encoding cyclin D1, and orthologs of p21 (p21-like) and retinoblastoma (Rb-like 1). To investigate the regulation of this process further, the translation of ccnd1 mRNA, a G1/S checkpoint control element, was impaired by microinjection of ccnd1-specific morpholino phosphorodiamidate (MO) 20mer or hydroxyprolyl-phosphono peptide nucleic acid (HypNA-pPNA) 16mer antisense oligonucleotides. The resulting downregulation of cyclin D1 protein resulted in microophthalmia and microcephaly, but not lethality. The phenotypes were not seen with 3-mismatch MO 20mers or 1-mismatch HypNA-pPNA 16mers, and were rescued by an exogenous ccnd1 mRNA construct with five mismatches. Collectively, these results indicate that transcription of key molecular determinants of asynchronous cell cycle control in zebrafish embryos commences at MBT and that the reduction of cyclin D1 expression compromises zebrafish eye and head development.
机译：在早期斑马鱼（Danio rerio）发育期间，直到受精后3 h囊胚中期过渡（MBT）才开始合子转录。 MBT专门划定了从S和M相的同步短单元格周期过渡到包含G1和G2相的完整周期的过渡。在这些阶段的转录谱分析和RT-PCR分析使我们能够确定这种转变与S / M期细胞周期控制基因（例如ccna2，ccnb1，ccnb2和ccne）的转录水平降低以及编码cyclin的ccnd1转录水平升高有关D1，以及p21（p21样）和成视网膜细胞瘤（Rb样1）的直系同源物。为了进一步研究该过程的调控，通过显微注射ccnd1特异性吗啉代二氨基磷酸二酰胺（MO）20mer或羟脯氨酰基膦酰基肽核酸（HypNA-pPNA）16mer来破坏G1 / S检查点控制元件ccnd1 mRNA的翻译。反义寡核苷酸。细胞周期蛋白D1蛋白的下调导致小眼症和小头畸形，但没有致死性。该表型在3-mismatch MO 20mers或1-mismatch HypNA-pPNA 16mers中未见，并由具有五个错配的外源ccnd1 mRNA构建物挽救。总的来说，这些结果表明斑马鱼胚胎中异步细胞周期控制的关键分子决定子的转录始于MBT，而细胞周期蛋白D1表达的降低会损害斑马鱼的眼睛和头部发育。

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