【摘要】：光學成像與圖像處理結合是認識生物結構空間構象的重要技術手段。而生物結構的空間構象對認識其生理功能,發(fā)育機制有主要意義�；诿庖邿晒夥椒�,本文采用雙光子熒光顯微成像技術,原位且立體地獲取生物樣品圖像。進一步,本文通過圖像三維分割、三維重構及數(shù)字化建模,量化表征生物結構的構象信息。采用該方法,本文研究了卵母細胞紡錘體和染色體的空間構象。染色體的空間構象與染色體的密度,基因轉錄活躍程度,以及細胞核的功能有重要關聯(lián)。紡錘體的空間構象與染色體的排列與分離以及細胞分裂有關,其完整性對細胞能否正常繁殖有重要作用。卵母細胞中紡錘體和染色體的構象研究對評估卵母細胞質量,分析生殖發(fā)育問題有重要意義(如出生缺陷問題)。基于體外培養(yǎng)技術,本文首先研究了氨甲喋呤對卵母細胞紡錘體和染色體的影響。氨甲喋呤是一種抗葉酸代謝類藥物,具有抑制細胞增殖的功能,廣泛用于臨床醫(yī)治惡性腫瘤、類風濕性關節(jié)炎及異位妊娠等疾病。許多研究證明,氨甲喋呤對快速分裂的體細胞很強的副作用。然而,其對女性生殖細胞的影響尚不明確。本文研究發(fā)現(xiàn)與對照組(84%)相比,生發(fā)泡破裂比率在氨甲喋呤組下降至73%(p0.05)。氨甲喋呤組的第一極體排出率(53%)也明顯低于對照組(63%;p0.05)。氨甲喋呤組染色體異常排布比率和紡錘體形態(tài)異常比率分別為60%和49%,均明顯高于對照組情況(24%,17%;p0.05)。以上結果提示氨甲喋呤可影響卵母細胞的成熟質量。隨后,本文又研究了激素超排對卵母細胞質量的影響。激素超數(shù)排卵作為一種非自然排卵方式,在臨床輔助生殖及基礎科學研究中應用廣泛。但此種排卵方式對卵母細胞成熟質量的影響仍不明確。我們研究發(fā)現(xiàn),激素超排造成染色體空間排布異常,但不影響染色體的體積、表面積等幾何構型。此結果可能提示激素超排干擾了染色體的運動與遷移功能。本論文建立了一套數(shù)字化表征紡錘體和染色體空間構象的方法,研究結果對認識生殖發(fā)育問題有重要意義。
[Abstract]:The combination of optical imaging and image processing is an important technique for understanding the spatial conformation of biological structures. The spatial conformation of biological structure is of great significance in understanding its physiological function and developmental mechanism. Based on the immunofluorescence method, two-photon fluorescence microscopic imaging technique is used to obtain in situ and stereoscopic images of biological samples. Furthermore, the conformational information of biological structure is quantitatively represented by 3D segmentation, 3D reconstruction and digital modeling. By using this method, the spatial conformation of oocyte spindles and chromosomes was studied. The spatial conformation of chromosomes is closely related to the density of chromosomes, the degree of transcriptional activity of genes, and the function of nuclei. The spatial conformation of the spindle is related to the arrangement and separation of chromosomes and cell division, and its integrity plays an important role in the normal reproduction of cells. The study of the conformation of spindle and chromosome in oocytes is of great significance in evaluating the quality of oocytes and analyzing reproductive development problems (such as birth defects). Based on in vitro culture, the effects of methotrexate on oocyte spindle and chromosome were studied. Methotrexate is a kind of anti-folic acid metabolite with the function of inhibiting cell proliferation and is widely used in clinical treatment of malignant tumor rheumatoid arthritis and ectopic pregnancy and other diseases. Many studies have shown that methotrexate has a strong side effect on rapidly dividing somatic cells. However, its effect on female germ cells is unclear. Compared with the control group (84%), the rupture rate of foaming decreased to 73% (p0.05) in methotrexate group. The first polar body excretion rate (53%) in methotrexate group was significantly lower than that in control group (63% p0.05). The chromosomal abnormal arrangement ratio and spindle morphology abnormality rate of methotrexate group were 60% and 49%, respectively, which were significantly higher than those in the control group (24%, 1717%, p0.05). These results suggest that methotrexate may affect the maturation quality of oocytes. Then, the effect of hormone superovulation on oocyte quality was studied. Hormone superovulation, as an unnatural ovulation, is widely used in clinical assisted reproduction and basic scientific research. However, the effect of ovulation on the maturation quality of oocytes is still unclear. We found that hormone superovulation causes abnormal spatial distribution of chromosomes, but does not affect the geometric configuration of chromosomes, such as volume and surface area. These results may suggest that hormone superovulation interferes with the movement and migration of chromosomes. In this paper, a method of digitally characterizing the spatial conformation of spindles and chromosomes is established, and the results are of great significance to the understanding of reproductive development.
【學位授予單位】：沈陽師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q343;TP391.41

【參考文獻】

相關期刊論文 前10條

1 朱越;高萬榮;郭英呈;;提高全場光學相干層析系統(tǒng)成像質量的方法[J];光學學報;2015年05期

2 李驚亞;鮑曉菁;;中國新生兒缺陷調查[J];決策探索(上半月);2013年05期

3 尚彥霞;解世朋;張銘連;周平坤;;DNA損傷反應性蛋白參與中心體調控[J];中國生物化學與分子生物學報;2011年11期

4 張興會;張世偉;宋先忱;郭丹;豆興堂;薛冰;常青;;幼齡母羊超數(shù)排卵及卵母細胞成熟效果分析[J];中國畜牧獸醫(yī);2011年11期

5 張眉芳;;衛(wèi)生部發(fā)布《中國婦幼衛(wèi)生事業(yè)發(fā)展報告(2011)》[J];醫(yī)學與社會;2011年10期

6 張恒慶;朱立南;杜洋;賈俊玲;譚琨;;藜ISSR-PCR反應體系的優(yōu)化[J];沈陽師范大學學報(自然科學版);2011年02期

7 毛萌;朱軍;;出生缺陷監(jiān)測研究現(xiàn)狀[J];實用兒科臨床雜志;2009年11期

8 路佩瑤;張春香;岳文斌;;超數(shù)排卵在哺乳動物中的應用[J];山西農業(yè)科學;2009年05期

9 朱曉輝;朱忠勇;;相差顯微鏡的原理、結構和臨床應用[J];臨床檢驗雜志;2007年04期

10 李奇慧,朱明學,張黎明,董兆君;硫芥誘導小鼠染色體畸變和微核形成作用的研究[J];第三軍醫(yī)大學學報;2004年19期

相關博士學位論文 前4條

1 袁一峰;SUMO-1在小鼠卵母細胞減數(shù)分裂中的作用及其分子機制研究[D];華中農業(yè)大學;2014年

2 趙鳳英;染色體三維空間構象的熒光量化表征[D];清華大學;2014年

3 田寧;雙光子成像量化觀測小鼠卵母細胞染色體三維構象[D];清華大學;2013年

4 鄧仕超;光學顯微鏡圖像處理技術及應用研究[D];天津大學;2010年

相關碩士學位論文 前5條

1 吳銀玲;小鼠超數(shù)排卵后胚胎質量評價[D];河北大學;2016年

2 朱曉萌;全場光學相干層析術高分辨率成像研究[D];南京理工大學;2014年

3 楊尋;3D-FISH圖像的量化處理和統(tǒng)計分析[D];清華大學;2013年

4 曹秀霞;哈克尼西棉細胞質雄性不育恢復基因SSR標記定位及利用[D];中國農業(yè)科學院;2012年

5 國玉蕊;采用偏振光顯微鏡分析透明帶在受精過程中的作用[D];重慶醫(yī)科大學;2010年

，

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