本文選題：斑馬魚(yú)　切入點(diǎn)：Gfi1.2　出處：《北京協(xié)和醫(yī)學(xué)院》2009年博士論文　論文類型：學(xué)位論文

【摘要】：目的：建立以斑馬魚(yú)為模式動(dòng)物的內(nèi)耳損傷模型,研究Gfi1在斑馬魚(yú)內(nèi)耳發(fā)育不同時(shí)間段的表達(dá)譜,以及耳毒性藥物對(duì)內(nèi)耳毛細(xì)胞損傷的發(fā)生機(jī)制。 材料和方法：(1).選用發(fā)育至4.3hpf、12hpf、24hpf、48hpf的斑馬魚(yú)胚胎,運(yùn)用原位雜交技術(shù)觀察Gfi1.2基因在各時(shí)期胚胎中的表達(dá)情況。(2)選用48hpf的斑馬魚(yú)胚胎,分別用0.25mM.0.50mM、0.75mM、1.00mM、1.50mM的順鉑溶液培養(yǎng)8小時(shí),用原位雜交技術(shù)觀察Gfil.2在經(jīng)過(guò)各種處理后胚胎中的表達(dá)情況。(3)選用48hpf的斑馬魚(yú)胚胎,用1.00mM的順鉑溶液培養(yǎng)4小時(shí)、8小時(shí)、12小時(shí),用原位雜交技術(shù)觀察Gfi1.2在經(jīng)過(guò)各種處理后胚胎中的表達(dá)情況。 結(jié)果：(1)Gfi1.2基因在發(fā)育至4.3hpf、12hpf、24hpf、48hpf的斑馬魚(yú)胚胎中均有明確表達(dá),在48hpf的胚胎的內(nèi)耳中有明顯的表達(dá)。(2)用不同濃度順鉑處理的斑馬魚(yú)胚胎經(jīng)過(guò)原位雜交,發(fā)現(xiàn)用越高濃度處理的胚胎,Gfil.2的表達(dá)越弱。(3)用相同濃度的順鉑處理的斑馬魚(yú)胚胎,處理時(shí)間越長(zhǎng),Gfil.2的表達(dá)越弱。 結(jié)論：(1)斑馬魚(yú)是一種新興的研究脊椎動(dòng)物發(fā)育的模式生物,由于它具有發(fā)育周期短、胚胎透明內(nèi)耳易于觀察、價(jià)格相對(duì)低廉等優(yōu)點(diǎn),因而成為一種研究耳聾基因及內(nèi)耳發(fā)育的優(yōu)秀模型。本次實(shí)驗(yàn)證明,用斑馬魚(yú)作為研究藥物對(duì)內(nèi)耳毛細(xì)胞損害的模型也是可行的。(2)Gfi1是一種轉(zhuǎn)錄抑制因子,它在小鼠內(nèi)耳毛細(xì)胞發(fā)育的過(guò)程中扮演著重要的角色,實(shí)驗(yàn)證明它的同源基因Gfil.2在斑馬魚(yú)內(nèi)耳中也有持續(xù)表達(dá),而且可以作為毛細(xì)胞損害的標(biāo)記物。(3)順鉑對(duì)內(nèi)耳毛細(xì)胞的損害具有濃度依賴性和時(shí)間依賴性。
[Abstract]:Aim: to establish an animal model of inner ear injury in zebrafish, and to study the expression profile of Gfi1 at different stages of development of zebrafish inner ear and the mechanism of ototoxic drugs on the damage of inner ear hair cells. Materials and methods zebrafish embryos developed to 4.3hpfn12hpfhfhfhfan24hpfan48hpf were used to observe the expression of Gfi1.2 gene in different stages of embryos by in situ hybridization. The zebrafish embryos of 48hpf were cultured in 0.25mM.0.50mM0.75mM 0.75mMU 1.50mm cisplatin solution for 8 hours, respectively. In situ hybridization technique was used to observe the expression of Gfil.2 in the embryos treated with various treatments. The zebrafish embryos with 48hpf were cultured in 1.00mm cisplatin solution for 4 hours, 8 hours and 12 hours, respectively. The expression of Gfi1.2 in embryos treated with in situ hybridization was observed. Results the Gfi1.2 gene was expressed in all zebrafish embryos which developed to 4.3hpfhpfl12hpfan24hpfan48hpf, and obviously expressed in the inner ear of 48hpf embryos) zebrafish embryos treated with different concentrations of cisplatin were hybridized in situ, and the zebrafish embryos with different concentrations of cisplatin were hybridized in situ, and the expression of Gfi1.2 gene was observed in the inner ear of 48hpf embryos. It was found that the weaker the expression of Gfil.2 was in the embryos treated with higher concentration of Cisplatin, the weaker the expression of Gfil.2 in the zebrafish embryos treated with the same concentration of cisplatin was. Conclusion zebrafish is a new model organism for studying the development of vertebrates, because it has the advantages of short developmental cycle, easy observation of transparent inner ear of embryo, and relatively low price. Therefore, it has become an excellent model to study the deafness gene and the development of the inner ear. This experiment proved that it is feasible to use zebrafish as a drug to study the damage of hair cells in the inner ear. It plays an important role in the development of mouse inner ear hair cells, and it has been proved that its homologous gene Gfil.2 is also expressed continuously in zebrafish inner ear. Cisplatin can be used as a marker of hair cell damage in a concentration-and time-dependent manner.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R764.3;R-332

【共引文獻(xiàn)】

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本文編號(hào)：1585394