【摘要】：花背蟾蜍(Bufo Raddei Strauch)是本實(shí)驗(yàn)室研究眼睛發(fā)育常用的兩棲類模式動(dòng)物。本文利用常規(guī)組織學(xué)技術(shù)、透射電鏡技術(shù)、掃描電鏡技術(shù)和免疫組織化學(xué)技術(shù)對(duì)花背蟾蜍視網(wǎng)膜進(jìn)行了如下研究：(1)視網(wǎng)膜形態(tài)結(jié)構(gòu)及胚后形態(tài)發(fā)育過程研究,(2)視網(wǎng)膜生理活動(dòng)與角膜誘導(dǎo)關(guān)系研究,(3)不同齡期蟾蜍視網(wǎng)膜增殖細(xì)胞分裂能力研究。 利用光鏡技術(shù)和電鏡技術(shù)對(duì)正常成體花背蟾蜍視網(wǎng)膜形態(tài)進(jìn)行了觀察,研究結(jié)果顯示,光鏡下花背蟾蜍視網(wǎng)膜具有脊椎動(dòng)物視網(wǎng)膜典型的十層結(jié)構(gòu)。透射電鏡下觀察花背蟾蜍視網(wǎng)膜結(jié)構(gòu)特點(diǎn)為：視網(wǎng)膜色素上皮細(xì)胞胞質(zhì)內(nèi)存在獨(dú)特的巨大吞噬體；在光感受器細(xì)胞內(nèi)節(jié)可觀察到橢球體與肌樣體結(jié)構(gòu),視錐細(xì)胞內(nèi)節(jié)未觀察到起濾色作用脂滴；光感受器細(xì)胞外節(jié)周圍可見輔助外節(jié)和花萼狀突起斷面；光感受器細(xì)胞的內(nèi)突部分與第二級(jí)神經(jīng)元形成突觸帶。掃描電鏡對(duì)視網(wǎng)膜色素上皮觀察可見排列整齊的色素細(xì)胞突起。視網(wǎng)膜斷面觀察可看到大量的視桿細(xì)胞和少數(shù)視錐細(xì)胞。視桿細(xì)胞外、內(nèi)節(jié)表面分布明顯的小葉間溝。圍繞在光感受器細(xì)胞內(nèi)節(jié)周圍可見Muller細(xì)胞突起形成的外界膜。利用GFAP抗體對(duì)視網(wǎng)膜切片進(jìn)行免疫組化染色發(fā)現(xiàn),Muller細(xì)胞是花背蟾蜍視網(wǎng)膜中唯一的膠質(zhì)細(xì)胞,視野中可見它沿著眼球半徑呈單層放射狀排列的橫斷面,Muller細(xì)胞主干終末膨大形成的圓錐型終足分布于內(nèi)界膜。 利用半薄切片技術(shù)及電鏡技術(shù)對(duì)花背蟾蜍26-39期(胚后發(fā)育期)蝌蚪視網(wǎng)膜發(fā)育過程觀察發(fā)現(xiàn),視網(wǎng)膜分層區(qū)域逐漸從中央向邊緣區(qū)擴(kuò)展。在26、27、28和35期蝌蚪視網(wǎng)膜的邊緣存在一逐漸閉合的裂孔結(jié)構(gòu)。透射電鏡觀察可見到26至39期色素細(xì)胞色素顆粒逐漸增加,突起逐漸伸長(zhǎng)。同時(shí)可觀察到光感受器細(xì)胞外節(jié)內(nèi)的膜盤增多,視錐細(xì)胞內(nèi)節(jié)特有的含脂質(zhì)色素脂滴逐漸形成過程。 用移植皮膚將蝌蚪角膜替換,移植皮膚在一定因素誘導(dǎo)下透明成為角膜即角膜誘導(dǎo)。利用眶內(nèi)注射河豚毒素阻斷蝌蚪視神經(jīng)生理活動(dòng),觀察不同時(shí)間點(diǎn)移植角膜中黑素色顆粒數(shù)量變化發(fā)現(xiàn),實(shí)驗(yàn)組角膜與對(duì)照組角膜黑素色顆粒都隨著時(shí)間延續(xù)逐漸減少,但實(shí)驗(yàn)組角膜透明過程顯著慢于對(duì)照組,兩者差異顯著(p0.05)。此結(jié)果說(shuō)明視網(wǎng)膜生理活動(dòng)是誘導(dǎo)角膜透明的直接因素之一。 為了觀察花背蟾蜍視網(wǎng)膜內(nèi)神經(jīng)細(xì)胞增殖能力與蟾蜍年齡關(guān)系,利用Skeletochronology技術(shù)對(duì)花背蟾蜍年齡進(jìn)行了鑒定,并對(duì)其群體年齡結(jié)構(gòu)和生長(zhǎng)模式進(jìn)行了分析,分析結(jié)果顯示實(shí)驗(yàn)中所用的成體花背蟾蜍年齡可達(dá)到6齡。利用BrdU標(biāo)記分裂細(xì)胞的方法,對(duì)花背蟾蜍30、35、39期蝌蚪、變態(tài)后幼體,以及1齡、2齡、3齡成體視網(wǎng)膜增殖細(xì)胞進(jìn)行顯示表明,隨著花背蟾蜍生長(zhǎng)時(shí)間延續(xù),其視網(wǎng)膜中具有分裂能力的神經(jīng)細(xì)胞逐漸減少,2齡期視網(wǎng)膜中幾乎沒有增殖細(xì)胞存在。由此可以推論無(wú)尾兩棲動(dòng)物可能是視網(wǎng)膜神經(jīng)細(xì)胞由終生保持分裂能力到短期保持分裂能力的過渡動(dòng)物。 本研究從以上3方面對(duì)花背蟾蜍視網(wǎng)膜進(jìn)行觀察研究,在視網(wǎng)膜形態(tài)結(jié)構(gòu)方面觀察到色素上皮細(xì)胞中存在著巨大吞噬體、在胚后發(fā)育中觀察到的視網(wǎng)膜裂孔、以及神經(jīng)細(xì)胞分裂能力與年齡的關(guān)系、視網(wǎng)膜生理活動(dòng)與角膜誘導(dǎo)關(guān)系等,為今后進(jìn)一步進(jìn)行相關(guān)研究奠定了研究基礎(chǔ)。
[Abstract]:Bufo Raddei Strauch (Bufo Raddei Strauch) is one of the most commonly used animal models for the development of eye development in our laboratory. In this paper, the retinal morphology and morphology development were studied by using conventional histological techniques, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and immunohistochemistry. (2) Study on the relationship between retinal physiology activity and cornea induction, and (3) study on cell division ability of toad retina at different age. The morphology of toad retina was observed by light microscopy and electron microscopy. The results showed that the retinas of toad retina was the typical ten in the retina under light microscope. The structure of toad retina was observed under transmission electron microscope (TEM). An auxiliary outer section and a flower-shaped protrusion section are visible around the outer section of the photoreceptor cell; the inner and second stage neurons of the photoreceptor cell are formed Synaptic band. Scanning electron microscope showed the visible arrangement of pigment fine in retinal pigment epithelium. The observation of retinal cross-section can see a large number of rod cells and a few eyes. In addition to the cells of the cone, the surface distribution of the inner section was significantly smaller than that of the rod cells. An inter-leaf groove formed by a Muller cell protrusion that is visible around the inside of a photoreceptor cell. It was found by immunohistochemical staining of retinal sections with anti-inflammatory antibodies, which was the only glial cell in the retina of Bufo gartoads, and it was seen in the field of view that it was arranged radially along the radius of the eyeball. In cross-section, the cone-type final foot formed by the terminal enlargement of the main stem of Muller cells was distributed in Using semi-thin slice technique and electron microscope technique to observe the development of tadpole's retina during the 26-39 period (embryonic development stage), the retinal delamination area was gradually observed. In the 26, 27, 28, and 35 tadpoles, there was a gradual increase in the edge of the tadpoles retina. According to transmission electron microscope, the pigment particles of 26 to 39 pigment cells were gradually increased. At the same time, it can be observed that the film disc in the outer section of photoreceptor cells is increased, and the specific lipid-containing pigment grease is peculiar to the inside of the cone cell. The process is gradually formed. The tadpole cornea is replaced by transplanting the skin, and the transplanted skin is induced by a certain factor. Using intraorbital injection of dodotoxin to block tadpole's optic nerve physiological activity, the changes of the number of black pigment particles in the cornea were observed in different time points, and the corneal and black pigment particles in the experimental group and the control group were observed. As time continues to decrease, the corneal transparency in the experimental group is significantly slower than that in the control group, both The difference was significant (p0.05). This result shows that the retinal physiology activity is induced One of the direct factors of corneal transparency was to observe the relationship between the proliferation of nerve cells and the age of toad in toad retina. The age structure and growth model of the body were analyzed, and the results showed that By using BrdU, the cells were displayed in 30, 35, 39 tadpoles, perverted larvae, and 1, 2, and 3-year-old retinal proliferative cells. It is shown that with the duration of the growth time of the toad toad, the nerve cells with splitting ability in the retina are gradually decreased, and the age of 2 years There is almost no proliferating cell in the retina of the retina. Thus, it is possible to deduce that the non-proliferative cells may be retinal nerve cells that remain divided by life for life. From the above three aspects, we observed the retinal morphology, observed the presence of giant phagocytes in the epithelial cells of the pigment epithelium, and observed the retina after the embryo development. The relationship between the splitting ability and age of nerve cells, the relationship between retinal physiology activity and cornea induction, etc.
【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】：2011
【分類號(hào)】：R774.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周立志,宋榆鈞;長(zhǎng)春花背蟾蜍春夏季種群生態(tài)研究[J];動(dòng)物學(xué)雜志;1997年06期

2 周立志,宋榆鈞;花背蟾蜍繁殖種群食物多樣性及營(yíng)養(yǎng)生態(tài)位的研究[J];動(dòng)物學(xué)雜志;1998年02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 郭玫,陳廣文,翟心慧;牛蛙視網(wǎng)膜誘導(dǎo)型一氧化氮合酶免疫組化定位[J];動(dòng)物學(xué)雜志;2002年01期

5 汪曉鵬,仝允栩;花背蟾蜍角膜早期發(fā)育中膠原合成、分泌和分布的研究[J];動(dòng)物學(xué)報(bào);1992年03期

6 王子仁,仝允栩;花背蟾蜍蝌蚪變態(tài)期角膜發(fā)育的研究[J];動(dòng)物學(xué)報(bào);1989年04期

7 李俊鳳,吳奇久;黑斑蛙光感受器計(jì)數(shù)和分類——掃描電鏡研究[J];動(dòng)物學(xué)報(bào);1989年02期

8 馮伯森,孫穎,仝允栩;花背蟾蜍角膜早期形態(tài)發(fā)生中膠原合成的放射自顯影研究[J];動(dòng)物學(xué)報(bào);1990年01期

9 王子仁,R.L.麥耶;河豚毒素對(duì)金魚再生的視網(wǎng)膜頂蓋投射模式的影響[J];動(dòng)物學(xué)報(bào);1995年03期

10 高建民,張彥定;崇安髭蟾視網(wǎng)膜的組織結(jié)構(gòu)[J];動(dòng)物學(xué)報(bào);1996年03期

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