本文選題：預(yù)定視網(wǎng)膜色素上皮 + 預(yù)定神經(jīng)視網(wǎng)膜��； 參考：《蘭州大學(xué)》2012年博士論文

【摘要】：脊椎動(dòng)物視網(wǎng)膜的發(fā)育是一個(gè)復(fù)雜的形態(tài)學(xué)和分子生物學(xué)過(guò)程。在由視杯向成熟的視網(wǎng)膜的發(fā)育過(guò)程中,存在著細(xì)胞分化和轉(zhuǎn)分化現(xiàn)象,但是在視泡發(fā)育成預(yù)定視網(wǎng)膜色素上皮(PRPE)和預(yù)定神經(jīng)視網(wǎng)膜(PNR)的過(guò)程中,相關(guān)的研究比較少,并且其在形態(tài)變化中所涉及的生物學(xué)機(jī)制還不清楚。本實(shí)驗(yàn)就是通過(guò)形態(tài)學(xué)和免疫組織化學(xué)實(shí)驗(yàn),研究視泡發(fā)育過(guò)程中細(xì)胞凋亡和分子分化的機(jī)制。 本實(shí)驗(yàn)以花背蟾蜍為材料,通過(guò)蘇木精-伊紅(HE)染色觀察視泡發(fā)育過(guò)程的組織形態(tài)學(xué)變化；然后通過(guò)TUNEL染色法觀察視泡發(fā)育過(guò)程中預(yù)定視網(wǎng)膜色素上皮和預(yù)定神經(jīng)視網(wǎng)膜中的細(xì)胞凋亡(apoptosis)。最后通過(guò)星形膠質(zhì)細(xì)胞膠質(zhì)原纖維酸性蛋白(GFAP)、神經(jīng)絲蛋白(RT97)和色素合成過(guò)程的催化酶—酪氨酸酶(Tyrosinase, Tyr)的免疫組織化學(xué)染色,研究視泡發(fā)育過(guò)程中的分子分化。 形態(tài)學(xué)染色結(jié)果發(fā)現(xiàn),在整個(gè)視泡發(fā)育時(shí)期,預(yù)定神經(jīng)視網(wǎng)膜和預(yù)定視網(wǎng)膜色素上皮組織結(jié)構(gòu)變化明顯。間腦兩側(cè)向外突出,形成視泡。然后視泡向上突起,最終與表皮外胚層接觸。預(yù)定視網(wǎng)膜色素上皮的厚度逐漸變薄,而預(yù)定神經(jīng)視網(wǎng)膜的厚度逐漸增加。同時(shí),發(fā)現(xiàn)細(xì)胞凋亡存在于整個(gè)視泡發(fā)育過(guò)程,但是凋亡細(xì)胞的密度隨著視泡發(fā)育進(jìn)程而逐漸降低。在同一發(fā)育時(shí)期內(nèi),腹部和前部預(yù)定神經(jīng)視網(wǎng)膜中凋亡細(xì)胞的密度在不同的發(fā)育模式中相對(duì)較高(P0.01)；背腹軸發(fā)育模式的視網(wǎng)膜凋亡細(xì)胞密度大于前后軸發(fā)育模式,而且預(yù)定視網(wǎng)膜色素上皮的凋亡細(xì)胞密度大于預(yù)定神經(jīng)視網(wǎng)膜中的凋亡細(xì)胞密度(P0.01)。 胚胎期的視網(wǎng)膜免疫組織化學(xué)染色發(fā)現(xiàn),GFAP和RT97表達(dá)水平的變化趨勢(shì)接近一致。視泡發(fā)育初期,GFAP和RT97在預(yù)定視網(wǎng)膜色素上皮中的表達(dá)水平很高,相反在預(yù)定神經(jīng)視網(wǎng)膜表達(dá)非常少。視泡發(fā)育末期,GFAP和RT97在預(yù)定神經(jīng)視網(wǎng)膜表達(dá)水平提高,而在預(yù)定視網(wǎng)膜色素上皮中降至最低值。在絕大多數(shù)發(fā)育時(shí)期,GFAP和RT97在腹部和前部PNR中的表達(dá)水平在不同的發(fā)育模式中相對(duì)比較高(P0.01)。GFAP和RT97抗體在視網(wǎng)膜前后軸發(fā)育模式中的表達(dá)水平要高于背腹軸發(fā)育模式中的表達(dá)水平(P0.01)。在視泡發(fā)育初期,Tyr在預(yù)定視網(wǎng)膜色素上皮的表達(dá)水平很低,而在預(yù)定神經(jīng)視網(wǎng)膜的表達(dá)水平卻很高。在隨后的發(fā)育時(shí)期內(nèi),Tyr在預(yù)定視網(wǎng)膜色素上皮的表達(dá)水平逐漸提高,而在預(yù)定神經(jīng)視網(wǎng)膜的表達(dá)水平降至最低值。在背腹軸發(fā)育模式中,Tyr在腹部預(yù)定視網(wǎng)膜色素上皮的表達(dá)水平相對(duì)較高(P0.01)。在視泡發(fā)育前期,Tyr在前部預(yù)定視網(wǎng)膜色素上皮的表達(dá)水平相對(duì)較高(P0.01);但是到了發(fā)育后期,Tyr在后部預(yù)定視網(wǎng)膜色素上皮的表達(dá)水平相對(duì)較高(P0.01)。與視泡形成初期相比,GFAP、RT97和Tyr在視網(wǎng)膜中的表達(dá)水平變化明顯(P0.01)。 本實(shí)驗(yàn)結(jié)果說(shuō)明細(xì)胞凋亡是兩棲動(dòng)物視泡發(fā)育過(guò)程中的一種普遍生物學(xué)現(xiàn)象,它存在于各個(gè)發(fā)育階段,并形成了空間和時(shí)間的變化模式。細(xì)胞凋亡現(xiàn)象在背腹軸發(fā)育模式中比較明顯,因此細(xì)胞凋亡的作用可能在視泡的背腹軸發(fā)育模式中更為重要。在同一發(fā)育時(shí)期,預(yù)定視網(wǎng)膜色素上皮中的凋亡細(xì)胞的密度要高于預(yù)定神經(jīng)視網(wǎng)膜中的凋亡的細(xì)胞密度,而且預(yù)定視網(wǎng)膜色素上皮的厚度變的比預(yù)定神經(jīng)視網(wǎng)膜薄,因此細(xì)胞凋亡可能是引起眼發(fā)育早期形態(tài)分化的原因之一。 GFAP、RT97和Tyr表達(dá)的差異性,說(shuō)明在視泡發(fā)育過(guò)程中,發(fā)生了分子分化。預(yù)定神經(jīng)視網(wǎng)膜和預(yù)定視網(wǎng)膜色素上皮來(lái)源于相同的組織,因此在眼發(fā)育早期,GFAP、RT97和Tyr同時(shí)表達(dá),但是它們的表達(dá)水平不同。數(shù)據(jù)顯示這三種蛋白不同的表達(dá)水平可能和預(yù)定神經(jīng)視網(wǎng)膜和預(yù)定視網(wǎng)膜色素上皮在視泡中不同的位置有關(guān)。這三種抗體的表達(dá)在視泡發(fā)育5-6小時(shí)后有個(gè)交叉點(diǎn),此時(shí)視泡正好與外胚層接觸。從分子分化的觀點(diǎn)上認(rèn)為,特定的分子分化起始于視泡和外胚層的接觸后,這三種抗體的表達(dá)顯示出視網(wǎng)膜發(fā)育過(guò)程中的不同細(xì)胞類型的分化。在這個(gè)交叉點(diǎn)后,神經(jīng)元、神經(jīng)膠質(zhì)細(xì)胞和色素細(xì)胞根據(jù)它們將來(lái)的功能而不是在視網(wǎng)膜中的位置,開(kāi)始表達(dá)一些特異性的蛋白質(zhì)。
[Abstract]:The development of vertebrate retina is a complex morphological and molecular biological process . In the course of development of visual cup to mature retina , there is a phenomenon of cell differentiation and differentiation , but in the course of development of retinal pigment epithelium ( PRPE ) and pre - defined nerve retina ( pnr ) , the related research is less , and its biological mechanism involved in morphological change is not clear . This experiment is to study the mechanism of cell apoptosis and molecular differentiation in the process of cell development through morphological and immunohistochemical experiments .

In this experiment , the morphological changes of the visual cell were observed by HE staining .
The apoptosis of the retinal pigment epithelium and the pre - determined retinal pigment epithelium was observed by TUNEL staining . Finally , the molecular differentiation in the process of cell development was studied by immunohistochemical staining of GFAP , RT97 and catalytic enzyme - tyrosinase ( Tyr ) in the process of pigment synthesis .

At the same time , the density of apoptotic cells was relatively high in different developmental patterns ( P0.01 ) .
The apoptotic cell density of the retinal pigment epithelium was greater than that in the pre - and posterior - axis development model , and the density of apoptotic cells was greater than that in the pre - determined retinal pigment epithelium ( P0.01 ) .

The expression levels of GFAP and RT97 in the pre - determined retinal pigment epithelium were higher than those in the development model of the dorsal axises ( P0.01 ) .

The results indicate that apoptosis is a common biological phenomenon in the development of cell apoptosis , which exists in various developmental stages and forms a pattern of spatial and temporal changes . Apoptosis is more important in the development of dorsal axises .

The difference between GFAP , RT97 and Tyr indicates that molecular differentiation occurs during the development of the cell . The expression levels of these three antibodies may be related to the different positions of the predetermined nerve retina and the predetermined retinal pigment epithelium in the development of the eye . The data show that the expression levels of these three proteins may be related to the different positions of the predetermined nerve retina and the predetermined retinal pigment epithelium in the cell . The expression of these three antibodies is shown to be the differentiation of different cell types in the development of the retina . After this crossing point , the neurons , glia and pigment cells begin to express some specific proteins based on their future function , rather than in the retina .

【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R774.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 葛瑞昌 ,馮伯森 ,仝允栩;花背蟾蜍(Bufo naddei Strauch)的早期胚胎發(fā)育及分期[J];蘭州大學(xué)學(xué)報(bào)(自然科學(xué)版);1982年04期

2 程欣,沈偉哉,鄭達(dá)人;細(xì)胞凋亡研究進(jìn)展[J];解剖學(xué)雜志;1998年04期

3 趙振芳;花背蟾蜍胚胎發(fā)育的初步觀察[J];動(dòng)物學(xué)雜志;1991年02期

4 許紅霞,糜漫天,朱俊東,周永,郎海濱,徐朝霞,陳卡;谷氨酸興奮毒性及�；撬岣深A(yù)對(duì)大鼠視網(wǎng)膜GFAP表達(dá)的影響[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2005年11期

相關(guān)碩士學(xué)位論文 前1條

1 韓亞鵬;花背蟾蜍（Bufo raddei）眼視泡的形態(tài)發(fā)生和發(fā)育過(guò)程中細(xì)胞凋亡的研究[D];蘭州大學(xué);2007年

，

本文編號(hào)：1882429