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海洋无脊椎动物卵母细胞质量评估:荧光光谱法的新方法。

Oocyte quality assessment in marine invertebrates: a novel approach by fluorescence spectroscopy.

机构信息

Department of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Naples, NA, Italy.

Department of Sciences, University of Basilicata, Potenza, PZ, Italy.

出版信息

Biol Res. 2022 Nov 12;55(1):34. doi: 10.1186/s40659-022-00403-4.

Abstract

BACKGROUND

The assessment of oocyte quality is, nowadays, a major challenge in aquaculture, oocyte cryopreservation, and environmental science. Oocyte quality is a determining factor in fertilization and embryo development; however, there is still a lack of rapid and sensitive cellular markers for its assessment. Currently, its estimation is predominantly based on morphological analysis, which is subjective and does not consistently reflect the developmental competence of the oocytes. Despite several recent studies investigating molecular markers related to oocyte quality, methods currently available for their determination pose various technical challenges and limitations. In this study, we developed a novel approach based on fluorescence spectroscopy to assess different intrinsic physiological parameters that can be employed to evaluate egg quality in marine invertebrates that are widely used as animal models such as sea urchins and mussels.

RESULTS

Different physiological parameters, such as viability, mitochondrial activity, intracellular ROS levels, plasma membrane lipid peroxidation, and intracellular pH, for egg quality evaluation have been successfully assessed in sea urchins and mussels by using specific fluorescent dyes and detecting the fluorescent signals in eggs through fluorescence spectroscopy.

CONCLUSIONS

Based on our findings, we propose these physiological markers as useful predictors of egg quality in marine invertebrates; they can be estimated rapidly, selectively, and sensitively by employing this novel approach, which, due to the speed of analysis, the low cost, and easy use can be considered a powerful analytical tool for the egg quality assessment.

摘要

背景

如今,卵子质量评估是水产养殖、卵子冷冻保存和环境科学领域的一项重大挑战。卵子质量是受精和胚胎发育的决定因素;然而,仍然缺乏快速和敏感的细胞标记物来评估它。目前,其评估主要基于形态分析,这种方法具有主观性,并且不能始终如一地反映卵子的发育能力。尽管最近有几项研究调查了与卵子质量相关的分子标记物,但目前用于确定这些标记物的方法存在各种技术挑战和限制。在这项研究中,我们开发了一种基于荧光光谱的新方法,用于评估不同的内在生理参数,这些参数可用于评估广泛用作动物模型的海洋无脊椎动物(如海胆和贻贝)的卵子质量。

结果

通过使用特定的荧光染料并通过荧光光谱检测卵子中的荧光信号,成功评估了海胆和贻贝中不同的生理参数,如活力、线粒体活性、细胞内 ROS 水平、质膜脂质过氧化和细胞内 pH 值,用于评估卵子质量。

结论

基于我们的发现,我们提出这些生理标记物作为海洋无脊椎动物卵子质量的有用预测指标;通过采用这种新方法,可以快速、选择性和敏感地估计它们,由于分析速度快、成本低且易于使用,因此可以被认为是评估卵子质量的有力分析工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a09/9652798/c078c7088396/40659_2022_403_Fig1_HTML.jpg

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