【摘要】： 在大腦發(fā)育過(guò)程中，細(xì)胞外的各種因子引導(dǎo)著神經(jīng)生長(zhǎng)錐向著最終的目的地延伸，從而形成精密而復(fù)雜的神經(jīng)網(wǎng)絡(luò)。但是其中的分子機(jī)制仍然有許多還不清楚。Semaphorin家族是一個(gè)重要的神經(jīng)導(dǎo)向因子家族，其中semaphorin-3F(Sema3F)是這個(gè)家族重要的成員，它在神經(jīng)系統(tǒng)發(fā)育過(guò)程中發(fā)揮重要作用。近來(lái)又發(fā)現(xiàn)Sema3F基因突變和腫瘤的形成密切相關(guān)，這使得進(jìn)一步研究這個(gè)導(dǎo)向分子的功能和信號(hào)轉(zhuǎn)導(dǎo)機(jī)理變得更有意義。 為了研究該分子在小腦發(fā)育中的作用，我們運(yùn)用了生長(zhǎng)錐塌縮實(shí)驗(yàn)，生長(zhǎng)錐轉(zhuǎn)向分析，以及小腦組織塊共培養(yǎng)方法，來(lái)觀察Sema3F對(duì)培養(yǎng)的大鼠小腦顆粒神經(jīng)元的軸突生長(zhǎng)和細(xì)胞遷移過(guò)程中的作用。并進(jìn)一步運(yùn)用藥理學(xué)實(shí)驗(yàn)來(lái)檢測(cè)該分子作用的下游信號(hào)機(jī)制。 我們發(fā)現(xiàn)Sema3F的高親和力受體neuropilin-2(Npn2)在小腦顆粒細(xì)胞中有高表達(dá)。在生長(zhǎng)錐轉(zhuǎn)向?qū)嶒?yàn)中，一個(gè)細(xì)胞外的Sema3F梯度能使培養(yǎng)的小腦顆粒神經(jīng)元的軸突發(fā)生Npn2依賴的吸引性的轉(zhuǎn)向。而且，在組織塊共培養(yǎng)中，，分泌Sema3F的細(xì)胞團(tuán)能使小腦顆粒細(xì)胞偏向它遷移。我們發(fā)現(xiàn)Sema3F的吸引性作用在cGMP信號(hào)通路被抑制時(shí)受阻斷，在細(xì)胞內(nèi)cGMP水平增加時(shí)被削弱。進(jìn)而在生長(zhǎng)錐塌縮實(shí)驗(yàn)中，我們發(fā)現(xiàn)Sema3F能減弱因抑制基礎(chǔ)水平的cGMP而引起的顆粒細(xì)胞生長(zhǎng)錐的塌縮反應(yīng)。 總而言之，本研究發(fā)現(xiàn)Sema3F通過(guò)激活cGMP對(duì)小腦顆粒細(xì)胞的生長(zhǎng)錐延伸和細(xì)胞遷移產(chǎn)生吸引性的導(dǎo)向作用。
[Abstract]:During the development of the brain, various extracellular factors lead the nerve growth cone to the final destination to form a sophisticated and complex neural network. However, many of the molecular mechanisms are still unclear. Semaphorin family is an important neuro-guiding factor family, in which semaphorin-3F (Sema3F) is an important member of this family, which plays an important role in the development of nervous system. Recently, it has been found that the mutation of Sema3F gene is closely related to tumor formation, which makes it more meaningful to further study the function and signal transduction mechanism of this guiding molecule. To study the role of the molecule in the development of the cerebellum, we used the growth cone collapse test, the growth cone shift analysis, and the co-culture of the cerebellar tissue mass. To observe the role of Sema3F in axonal growth and cell migration of cultured rat cerebellar granular neurons. Further pharmacological experiments were used to detect the downstream signaling mechanism of the molecule. We found that neuropilin-2 (Npn2), a high affinity receptor of Sema3F, is highly expressed in cerebellar granulosa cells. In the growth cone turning experiment, an extracellular Sema3F gradient can induce the Npn2 dependent attractiveness of the axons of cultured cerebellar granular neurons. Moreover, in tissue mass co-culture, the Sema3F secreting cell mass causes the cerebellar granulosa cells to migrate in favor of it. We found that the attraction of Sema3F was blocked when the cGMP signaling pathway was inhibited and weakened when the intracellular cGMP level increased. Furthermore, in the growth cone collapse experiment, we found that Sema3F can attenuate the collapse of granulosa cell growth cone caused by inhibition of cGMP at basic level. In conclusion, we found that Sema3F can induce the growth cone extension and cell migration of cerebellar granulosa cells by activating cGMP.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R33

【共引文獻(xiàn)】

相關(guān)期刊論文 前2條

1 黃倩;周雪;丁艷;彭謹(jǐn);趙偉;章為;;針刺對(duì)備用背根節(jié)神經(jīng)元NP-1表達(dá)的影響[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2006年03期

2 溫華知;江洪;魯志兵;胡笑容;吳兵;徐盛開;黃從新;;大鼠心肌梗死后梗死周邊區(qū)Sema 3A的變化及其對(duì)心室顫動(dòng)閾值的影響[J];中國(guó)心臟起搏與心電生理雜志;2009年05期

相關(guān)碩士學(xué)位論文 前3條

1 李佳多;Sema3A對(duì)小鼠骨髓來(lái)源的DC細(xì)胞表面分子B7-H3表達(dá)影響的實(shí)驗(yàn)研究[D];華中科技大學(xué);2011年

2 丁艷;部分去背根后脊髓內(nèi)Sema3A、NP-1及備用背根節(jié)內(nèi)NP-1表達(dá)的變化[D];四川大學(xué);2006年

3 黃倩;針刺對(duì)部分去背根貓DRG、脊髓背角中NP-1及Sema3A表達(dá)的影響[D];四川大學(xué);2006年

本文編號(hào)：2284031