小鼠卵母细胞质量的光谱研究

Spectral Study on Mouse Oocyte Quality

卵母细胞质量是影响哺乳动物体外受精能力、胚胎体外发育潜能以及妊娠成功与否的关键因素,显著影响着人类的辅助生殖、动物的育种、品种改良及保存濒危物种等工作,对其质量的研究具有重要的应用价值。目前常规的质量评估手段主要从形态学角度、从生化分析角度展开。但传统的基于卵母细胞形态甄选评估其发育质量的方法,其结果的可靠性较强地依赖于技术人员的经验,具有主观性较强的特点;新型的基于生物学技术检测生化指标的方法虽能弥补基于形态检测方法的不足但仍存在侵入式研究、生物操作步骤繁琐复杂、实验耗时长,影响卵母细胞后续发育的缺点。众所周知,生物组织异变往往伴随着内部生化组分的改变,而生化组分的改变往往表现出光谱的差异。鉴于此,利用紫外-可见分光光度计和自改建多光谱成像系统研究卵母细胞的光谱,并分析了未成熟与成熟卵母细胞、新鲜成熟与排卵后老化卵母细胞的光谱差别。研究发现,(1)与新鲜成熟卵母细胞的紫外-可见光谱(190~1 100 nm)相比,未成熟与排卵后老化卵母细胞的光谱均出现数个峰位的增减变化。未成熟卵母细胞与新鲜成熟卵母细胞的紫外-可见光谱相比,缺失了205和579 nm两个峰位以及894~941 nm的波段,增加了593 nm处的峰位;排卵后老化卵母细胞与新鲜成熟卵母细胞的紫外-可见光谱相比,缺失了205、 445和579 nm的三个峰位以及846~941 nm的波段,也增加了593 nm处的峰位。(2)可见光波段的多光谱成像数据显示,与新鲜成熟卵母细胞相比,未成熟卵母细胞各生物结构在451和467 nm处的透射光谱强度下降,排卵后老化卵母细胞各生物结构在425、 431、 571和669 nm处的透射光谱强度降低。综上可见(特别是新鲜成熟与老化卵母细胞之间在多光谱成像中展现的光谱差别),光谱检测在甄别卵母细胞质量方面展现出了巨大的可行性。

Oocyte quality is the key factor to affect the ability of in-vitro fertilization,the potential of embryo development in vitro and the success of pregnancy of mammals,there by significantly affecting the assisted reproduction of human beings,the breeding of animals,the improvement of varieties and the preservation of endangered species.The research on oocyte quality has important application value.Currently,the common quality assessment methods are mainly performed from the morphology and biochemistry analysis perspective.However,the traditional method of evaluating the development quality of oocytes based on morphological selection has the characteristics of strong subjectivity,and the reliability of the results strongly depends on technicians’experience.Although the new method of detecting biochemical indexes based on biological technology can make up for the shortcomings of morphological detection methods,there are still some shortcomings of invasive research,complicated biological operation steps,long experimental time-consuming and affecting the subsequent development of oocytes.As we all know,Abnormal changes in biological tissue are often accompanied by changes in its internal biochemical components,which often shows spectral differences.Thus,this paper used ultraviolet-visible(UV-Vis)spectrophotometer and self-modified multispectral imaging system to study the spectra of oocytes,and the spectral data were analyzed between immature and mature oocytes,fresh mature and postovulatory aged oocytes.It was found that(1)compared with the UV-Vis spectra(190~1100 nm)of fresh matured oocytes,the spectra of immature oocytes and aged oocytes showed several peak changes.The UV-Vis spectra of immature oocytes missed two peaks of 205 and 579 nm and the band of 894~941 nm and added the peak of 593nm compared with that of fresh matured oocytes.For the UV-Vis spectra of the aged oocytes,three peaks of 205,445 and 579 nm and the band of 846 to 941 nm disappeared and the peak at 593 nm also increased.(2)The visible-band multispectral imaging data showed that compared with fresh matured oocytes,the transmission spectrum intensity at various biological structures of immature oocytes decreased at 451 and 467 nm,and the corresponding data of aged oocytes decreased at 425,431,571 and 669 nm.In summary(specially,the spectral differences between fresh matured and aged oocytes in multispectral imaging),it is thus clear that spectral detection shows great feasibility in identifying oocyte quality.