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Effects of Lactobacillus rhamnosus on zebrafish oocyte maturation: an FTIR imaging and biochemical analysis

机译:鼠李糖乳杆菌对斑马鱼卵母细胞成熟的影响:FTIR成像和生化分析

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The aim of this study was to verify the effects of probiotic Lactobacillus rhamnosus on zebrafish oocyte maturation using FPA (focal plane array) FTIR imaging together with specific biochemical assays (SDS–PAGE, real-time PCR and enzymatic assay). Oocyte growth is prevalently due to a vitellogenic process which consists of the hepatic synthesis of vitellogenin and its selective uptake during maturation. The administration of L. rhamnosus IMC 501 for 10 days induced chemical changes to oocyte composition, promoting the maturation process. Some interesting biochemical features, linked to protein secondary structure (amide I band) and to phospholipidic and glucidic patterns, were detailed by vibrational analysis. The spectroscopic results were supported by the early increase of the lysosomal enzyme involved in the final oocyte maturation, the cathepsin L. This enzyme increases during follicle maturation, with the highest levels in class IV oocytes. In treated females, class III oocytes showed higher cathepsin L gene expression and enzymatic activity, with levels comparable to class IV oocytes isolated from controls; this can be related to the proteolytic cleavage of the higher molecular mass yolk protein components, as evidenced by SDS–PAGE.
机译:这项研究的目的是使用FPA(焦平面阵列)FTIR成像以及特定的生化测定(SDS-PAGE,实时PCR和酶促测定)来验证益生菌鼠李糖乳杆菌对斑马鱼卵母细胞成熟的影响。卵母细胞的生长主要归因于卵黄形成过程,该过程由卵黄原蛋白的肝脏合成及其在成熟过程中的选择性摄取组成。给予鼠李糖乳杆菌IMC 501 10天后,卵母细胞组成发生化学变化,从而促进了成熟过程。通过振动分析详细介绍了一些有趣的生化特征,这些特征与蛋白质的二级结构(酰胺I谱带)以及磷脂和糖质模式有关。参与最终卵母细胞成熟的溶酶体酶(组织蛋白酶L)的早期增加支持了光谱结果。该酶在卵泡成熟过程中增加,在IV类卵母细胞中含量最高。在接受治疗的雌性中,III类卵母细胞显示出较高的组织蛋白酶L基因表达和酶活性,其水平与从对照中分离的IV类卵母细胞相当。如SDS-PAGE所示,这可能与更高分子量的蛋黄蛋白成分的蛋白水解裂解有关。

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