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

【摘要】：脊椎動物視網(wǎng)膜的發(fā)育是一個復雜的形態(tài)學和分子生物學過程。在由視杯向成熟的視網(wǎng)膜的發(fā)育過程中,存在著細胞分化和轉(zhuǎn)分化現(xiàn)象,但是在視泡發(fā)育成預定視網(wǎng)膜色素上皮(PRPE)和預定神經(jīng)視網(wǎng)膜(PNR)的過程中,相關的研究比較少,并且其在形態(tài)變化中所涉及的生物學機制還不清楚。本實驗就是通過形態(tài)學和免疫組織化學實驗,研究視泡發(fā)育過程中細胞凋亡和分子分化的機制。 本實驗以花背蟾蜍為材料,通過蘇木精-伊紅(HE)染色觀察視泡發(fā)育過程的組織形態(tài)學變化；然后通過TUNEL染色法觀察視泡發(fā)育過程中預定視網(wǎng)膜色素上皮和預定神經(jīng)視網(wǎng)膜中的細胞凋亡(apoptosis)。最后通過星形膠質(zhì)細胞膠質(zhì)原纖維酸性蛋白(GFAP)、神經(jīng)絲蛋白(RT97)和色素合成過程的催化酶—酪氨酸酶(Tyrosinase, Tyr)的免疫組織化學染色,研究視泡發(fā)育過程中的分子分化。 形態(tài)學染色結(jié)果發(fā)現(xiàn),在整個視泡發(fā)育時期,預定神經(jīng)視網(wǎng)膜和預定視網(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。預定視網(wǎng)膜色素上皮的厚度逐漸變薄,而預定神經(jīng)視網(wǎng)膜的厚度逐漸增加。同時,發(fā)現(xiàn)細胞凋亡存在于整個視泡發(fā)育過程,但是凋亡細胞的密度隨著視泡發(fā)育進程而逐漸降低。在同一發(fā)育時期內(nèi),腹部和前部預定神經(jīng)視網(wǎng)膜中凋亡細胞的密度在不同的發(fā)育模式中相對較高(P0.01)；背腹軸發(fā)育模式的視網(wǎng)膜凋亡細胞密度大于前后軸發(fā)育模式,而且預定視網(wǎng)膜色素上皮的凋亡細胞密度大于預定神經(jīng)視網(wǎng)膜中的凋亡細胞密度(P0.01)。 胚胎期的視網(wǎng)膜免疫組織化學染色發(fā)現(xiàn),GFAP和RT97表達水平的變化趨勢接近一致。視泡發(fā)育初期,GFAP和RT97在預定視網(wǎng)膜色素上皮中的表達水平很高,相反在預定神經(jīng)視網(wǎng)膜表達非常少。視泡發(fā)育末期,GFAP和RT97在預定神經(jīng)視網(wǎng)膜表達水平提高,而在預定視網(wǎng)膜色素上皮中降至最低值。在絕大多數(shù)發(fā)育時期,GFAP和RT97在腹部和前部PNR中的表達水平在不同的發(fā)育模式中相對比較高(P0.01)。GFAP和RT97抗體在視網(wǎng)膜前后軸發(fā)育模式中的表達水平要高于背腹軸發(fā)育模式中的表達水平(P0.01)。在視泡發(fā)育初期,Tyr在預定視網(wǎng)膜色素上皮的表達水平很低,而在預定神經(jīng)視網(wǎng)膜的表達水平卻很高。在隨后的發(fā)育時期內(nèi),Tyr在預定視網(wǎng)膜色素上皮的表達水平逐漸提高,而在預定神經(jīng)視網(wǎng)膜的表達水平降至最低值。在背腹軸發(fā)育模式中,Tyr在腹部預定視網(wǎng)膜色素上皮的表達水平相對較高(P0.01)。在視泡發(fā)育前期,Tyr在前部預定視網(wǎng)膜色素上皮的表達水平相對較高(P0.01);但是到了發(fā)育后期,Tyr在后部預定視網(wǎng)膜色素上皮的表達水平相對較高(P0.01)。與視泡形成初期相比,GFAP、RT97和Tyr在視網(wǎng)膜中的表達水平變化明顯(P0.01)。 本實驗結(jié)果說明細胞凋亡是兩棲動物視泡發(fā)育過程中的一種普遍生物學現(xiàn)象,它存在于各個發(fā)育階段,并形成了空間和時間的變化模式。細胞凋亡現(xiàn)象在背腹軸發(fā)育模式中比較明顯,因此細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)育時期,預定視網(wǎng)膜色素上皮中的凋亡細胞的密度要高于預定神經(jīng)視網(wǎng)膜中的凋亡的細胞密度,而且預定視網(wǎng)膜色素上皮的厚度變的比預定神經(jīng)視網(wǎng)膜薄,因此細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表達的差異性,說明在視泡發(fā)育過程中,發(fā)生了分子分化。預定神經(jīng)視網(wǎng)膜和預定視網(wǎng)膜色素上皮來源于相同的組織,因此在眼發(fā)育早期,GFAP、RT97和Tyr同時表達,但是它們的表達水平不同。數(shù)據(jù)顯示這三種蛋白不同的表達水平可能和預定神經(jīng)視網(wǎng)膜和預定視網(wǎng)膜色素上皮在視泡中不同的位置有關。這三種抗體的表達在視泡發(fā)育5-6小時后有個交叉點,此時視泡正好與外胚層接觸。從分子分化的觀點上認為,特定的分子分化起始于視泡和外胚層的接觸后,這三種抗體的表達顯示出視網(wǎng)膜發(fā)育過程中的不同細胞類型的分化。在這個交叉點后,神經(jīng)元、神經(jīng)膠質(zhì)細胞和色素細胞根據(jù)它們將來的功能而不是在視網(wǎng)膜中的位置,開始表達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 .

【學位授予單位】：蘭州大學
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R774.1

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本文編號：1882429