Mesoglein Expression During Aurelia aurita Life Cycle

A. V. Kotova, A. Naiden, Alex, er Shumeev, T. Shaposhnikova, O. Podgornaya, L. Adonin
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Abstract

Cnidarian is thought to possess two tissue layers: endoderm (gastroderm) and ectoderm, which are separated by the layer of extracellular matrix (ECM) called mesoglea. Aurelia aurita complex life cycle consists of several stages including alternating generations of sexual adult stage medusa and asexual stage polyp (scyphistoma). The main difference between polyp and medusa is the degree of the ECM (mesoglea) development. The new protein “mesoglein” was determined as one of the main components of mesoglea. Mesoglein is the component of the mesoglea “elastic” fibers. Previously we found that according to reverse transcription PCR mesoglein is expressed in the mature medusa exclusively in the mesogleal cells. The aim of the current work was to find out at what stage of development the specific mesoglein expression occurs. Mesogleins’ expression have been checked by PCR with specific primers on the template of transcriptomes’ cDNA from different stages; by mesogleins’ antibody staining on SDSPAGE and on paraffin sections; by histochemistry staining. In A.aurita life cycle mesoglein synthesis begins at scyphystoma polyp stage at RNA and protein level in mesogleal cells (Mc) which separated at this stage as the cell type. Mesoglein stored in granules both in Mc of adult medusa and polyp on its’ way to ECM elastic fibers. Our data perfectly correspond to the recently reported de novo transcriptome assembled from Illumina RNA-Seq data generated from six stages throughout the Aurelia life cycle.
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小红花生命周期中中胶蛋白的表达
刺胞动物被认为具有两层组织:内胚层(胃胚层)和外胚层,它们被称为中胶层的细胞外基质层(ECM)分开。auria aurita复杂的生命周期包括有性成虫期水母和无性期息肉(鞘瘤)交替的几个阶段。水螅和水母的主要区别在于中胶层的发育程度。新蛋白“中胶蛋白”被确定为中胶层的主要成分之一。中胶蛋白是中胶“弹性”纤维的组成部分。在此之前,我们通过反转录PCR发现,中胶蛋白在成熟水母中只在中胶膜细胞中表达。当前工作的目的是找出特定的中胶蛋白表达发生在发育的哪个阶段。在不同时期的转录组cDNA模板上,用特异引物PCR检测中胶蛋白的表达;在SDSPAGE和石蜡切片上进行中胶蛋白抗体染色;通过组织化学染色。在aurita的生命周期中,中胶蛋白的合成开始于鞘瘤息肉期,中胶细胞(Mc)的RNA和蛋白质水平在此阶段作为细胞类型分离。中胶蛋白在成体水母和水螅体中以颗粒形式储存于鞘膜弹性纤维中。我们的数据与最近报道的从头转录组完全一致,这些转录组是由Illumina RNA-Seq数据组装而成的,这些数据来自整个Aurelia生命周期的六个阶段。
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