本文選題：先天性靜止性夜盲 + 多電極陣列��； 參考：《第四軍醫(yī)大學(xué)》2013年博士論文

【摘要】：先天性靜止性夜盲(congenital stationary night blindness, CSNB)是一種非進(jìn)展性、以視桿視覺信號(hào)系統(tǒng)功能障礙為主的遺傳性視網(wǎng)膜病。我們實(shí)驗(yàn)室發(fā)現(xiàn)的Cacna1f基因自然突變CSNB大鼠,其電生理及行為學(xué)證明其克服了基因工程誘導(dǎo)突變動(dòng)物模式與臨床實(shí)際差異較大的缺陷,因此可以作為研究CSNB發(fā)病機(jī)制及視網(wǎng)膜信號(hào)通路之間相互關(guān)系的一種有效模型。Cav1.4型鈣通道為L-型鈣通道的亞型之一,其主要的功能單位a1f亞基由Cacnalf基因編碼。目前發(fā)現(xiàn)Cacna1f基因多種位點(diǎn)突變可損傷視桿和視錐信號(hào)傳導(dǎo)通路,并與性染色體遺傳的不完全型靜止性夜盲密切相關(guān)。研究已證實(shí)Cacna1f基因突變導(dǎo)致的α1f亞基缺陷可使光感受器細(xì)胞帶狀突觸數(shù)量、形態(tài)以及功能發(fā)生異常,從而引發(fā)光感受器細(xì)胞與二級(jí)神經(jīng)元之間視覺信號(hào)傳導(dǎo)障礙。 近期形態(tài)學(xué)研究發(fā)現(xiàn),自然突變CSNB大鼠視網(wǎng)膜雙極細(xì)胞所發(fā)出的異常突觸可伸及外核層外部,外叢狀層厚度減少,水平細(xì)胞數(shù)目減少,而視網(wǎng)膜內(nèi)層的超微結(jié)構(gòu)則相對(duì)正常完整。上述視網(wǎng)膜外層發(fā)生的形態(tài)學(xué)變化很可能影響視網(wǎng)膜內(nèi)層的視覺信號(hào)通路的傳導(dǎo)整合功能。然而,CSNB動(dòng)物視網(wǎng)膜外層所發(fā)生的形態(tài)學(xué)變化及神經(jīng)遞質(zhì)釋放功能障礙對(duì)視網(wǎng)膜最內(nèi)層的神經(jīng)節(jié)細(xì)胞會(huì)產(chǎn)生何種病理學(xué)效應(yīng),至今未見諸于報(bào)道。 多電極陣列系統(tǒng)已廣泛應(yīng)用于視網(wǎng)膜神經(jīng)元電生理活動(dòng)記錄等實(shí)驗(yàn)研究,由于其具備完整性、非侵入性的優(yōu)點(diǎn),因而此電生理實(shí)驗(yàn)技術(shù)彌補(bǔ)了傳統(tǒng)的活體外記錄與復(fù)雜的活體內(nèi)記錄的先天缺陷,目前已經(jīng)被應(yīng)用于記錄監(jiān)測(cè)光感受器細(xì)胞、神經(jīng)節(jié)細(xì)胞等視網(wǎng)膜神經(jīng)元的生理活動(dòng)。本研究采用多電極陣列記錄系統(tǒng)和先天性自然突變大鼠動(dòng)物模型,根據(jù)其視網(wǎng)膜內(nèi)層信號(hào)通路的視覺電生理信號(hào)所發(fā)生的特征性改變來證明其信號(hào)通路發(fā)生的重塑,并以此為基礎(chǔ)闡明通路重塑的發(fā)生機(jī)制,為進(jìn)一步制訂治療方案及采取針對(duì)性的視覺干預(yù)保護(hù)措施提供依據(jù)。 材料與方法 實(shí)驗(yàn)采用近交系CSNB大鼠(SPF級(jí),F23,8-10周齡)及正常同齡對(duì)照Wt大鼠,將大鼠暗適應(yīng)12-15小時(shí),深度麻醉后處死大鼠摘取眼球,除去眼前節(jié)和視網(wǎng)膜色素上皮層,制備視網(wǎng)膜鋪片,并將其神經(jīng)節(jié)細(xì)胞面向下貼附于多電極陣列記錄芯片,并使其適應(yīng)生物記錄芯片中的灌流環(huán)境。藥理學(xué)實(shí)驗(yàn)環(huán)節(jié)中,實(shí)驗(yàn)中所涉及的藥品經(jīng)活性灌注系統(tǒng)進(jìn)入灌流液。電腦刺激器控制的發(fā)光二級(jí)管所產(chǎn)生的白光經(jīng)透鏡系統(tǒng)匯聚形成光斑,投射到生物芯片灌流槽之中的視網(wǎng)膜鋪片上,給予其同步全野光刺激。多電極陣列系統(tǒng)電極所采集到的神經(jīng)元電信號(hào)經(jīng)過放大器過濾并放大,存儲(chǔ)于電腦系統(tǒng)供后續(xù)離線軟件分析處理。單電極所采集的電生理信號(hào)進(jìn)行放電分析時(shí),其檢測(cè)閾值設(shè)定為平均幅值的四倍標(biāo)準(zhǔn)差。然后應(yīng)用Offline Sorter將備選放電波形進(jìn)行處理,采取其前三個(gè)主要組成部分應(yīng)用NeuroExplore軟件進(jìn)行后續(xù)分析：使用K-均值分解類聚算法識(shí)別集簇放電；分析單個(gè)記錄單元的放電序列間距直方圖來確定其放電的規(guī)則性及峰電位分布,并結(jié)合單個(gè)記錄單元的圍刺激時(shí)間柱形圖來判斷放電細(xì)胞的種類。電刺激實(shí)驗(yàn)環(huán)節(jié)中,位于陣列中央的28號(hào)電極所產(chǎn)生的雙階平衡電流刺激序列誘發(fā)視網(wǎng)膜神經(jīng)元電生理反應(yīng),并為陣列中其它電極采集記錄,應(yīng)用相同軟件進(jìn)行分析處理。 結(jié)果 1.野生型SD大鼠與CSNB大鼠視網(wǎng)膜光刺激誘導(dǎo)場(chǎng)電位 施加光刺激程序后,多電極陣列記錄系統(tǒng)可從Wt大鼠視網(wǎng)膜鋪片采集到典型的光誘發(fā)場(chǎng)電位波形,其a波峰值為0.196±0.038mv(n-20)。應(yīng)用ON-型雙極細(xì)胞mGluR6受體的特異性拮抗劑APB (100μM/L)不能阻斷a-波卻可阻斷b-波。CSNB大鼠視網(wǎng)膜鋪片記錄中,光誘發(fā)場(chǎng)電位a-波峰值顯著減小(0.078±0.026mv, n-20, P0.01), b-波缺如。 2.野生型SD大鼠與CSNB大鼠視網(wǎng)膜電刺激放電反應(yīng) 施加雙階平衡電流刺激后,在野生型大鼠和CSNB大鼠電刺激后原始波形中均可發(fā)現(xiàn)兩種類型電刺激反應(yīng)：短潛伏期反應(yīng)和長潛伏期反應(yīng)。按照神經(jīng)節(jié)細(xì)胞與電流刺激電極的間距將其分為14組：(100μm組,141μm組,200μm組,223μm組,283μm組,300μm組,316μm組,361μm組,387μm組,400μm組,424pm組,447μm組,500μmm組和566μm組)。結(jié)果發(fā)現(xiàn)神經(jīng)節(jié)細(xì)胞的電刺激反應(yīng)強(qiáng)度隨其與位于陣列中央的電流刺激電極間的距離增加而降低；在所分組中,CSNB大鼠視網(wǎng)膜的長潛伏期反應(yīng)的放電數(shù)目均較野生型大鼠減少(P0.01)。 3.野生型SD大鼠與CSNB大鼠視網(wǎng)膜自發(fā)性放電反應(yīng) 多電極陣列記錄系統(tǒng)從每個(gè)視網(wǎng)膜鋪片中可同時(shí)記錄到來自30個(gè)-90個(gè)神經(jīng)節(jié)細(xì)胞的自發(fā)放電。分析發(fā)現(xiàn)CSNB大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的自發(fā)放電頻率(9.93±4.14Hz,n=386)顯著高于野生型大鼠(2.610.73Hz,n=295, P0.01)。從累積自發(fā)放電頻率曲線圖可以觀察到,CSNB大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的自發(fā)放電頻率曲線較野生型大鼠斜率降低且發(fā)生右移。 4.野生型SD大鼠與CSNB大鼠視網(wǎng)膜自發(fā)性放電神經(jīng)節(jié)細(xì)胞的類型 將產(chǎn)生自發(fā)性放電反應(yīng)的神經(jīng)節(jié)細(xì)胞歸類為四種類型：爆發(fā)型放電細(xì)胞,常規(guī)型放電細(xì)胞,非常規(guī)型放電細(xì)胞,混合型放電細(xì)胞。CSNB大鼠視網(wǎng)膜爆發(fā)型放電細(xì)胞和混合型放電細(xì)胞占所有自發(fā)放電神經(jīng)節(jié)細(xì)胞的比重高于野生型大鼠(Wt：爆發(fā)型15.2%,混合型：24.7%,n-20；CSNB:爆發(fā)型：24.3%,混合型：31.6%,n-20,P0.01)而非常規(guī)型放電細(xì)胞占所有自發(fā)放電神經(jīng)節(jié)細(xì)胞的比重低于野生型大鼠(Wt:51.4%, CSNB:36.8%; P0.01)。 CSNB大鼠視網(wǎng)膜混合型放電神經(jīng)節(jié)細(xì)胞的放電總數(shù)(246553個(gè),n-20)顯著高于Wt大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞混合混合型放電細(xì)胞的放電總數(shù)(112089,n=20,P0.01)。 CSNB大鼠和Wt大鼠視網(wǎng)膜其它三種神經(jīng)節(jié)細(xì)胞的放電總數(shù)無顯著差異(P0.05)。 5.野生型SD大鼠與CSNB大鼠視網(wǎng)膜光誘導(dǎo)放電反應(yīng) 按照對(duì)光反應(yīng)的放電特征可將來自CSNB大鼠和Wt大鼠視網(wǎng)膜的神經(jīng)節(jié)細(xì)胞分為六大類：1)給光型(ON-型)；2)撤光型(OFF-型)；3)給-撤光型(ON-OFF型)；4)持續(xù)給光型(sustained ON型)；5)持續(xù)給-撤光型(sustained ON-OFF型)；6)延遲撤光型(delayed OFF型)。CSNB大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞光誘導(dǎo)放電反應(yīng)的平均頻率(16.40±9.97Hz,n-340)較野生型大鼠顯著降低(38.6211.78Hz,n=373, P0.01)。與Wt大鼠視網(wǎng)膜相比,CSNB大鼠視網(wǎng)膜的暗適應(yīng)自發(fā)放電(背景噪聲)頻率較高而光誘導(dǎo)放電反應(yīng)(視覺信號(hào))頻率較低,因此其信號(hào)/噪聲比(S-N ration)減小。 6.野生型SD大鼠與CSNB大鼠視網(wǎng)膜自發(fā)放電的互相關(guān)活動(dòng) 在野生型大鼠視網(wǎng)膜中,觀察到三種特征性神經(jīng)節(jié)細(xì)胞互相關(guān)活動(dòng)類型：短時(shí)程互相關(guān)活動(dòng)、中時(shí)程互相關(guān)活動(dòng)和長時(shí)程互相關(guān)活動(dòng)。而在CSNB大鼠視網(wǎng)膜記錄數(shù)據(jù)中只觀察到短時(shí)程互相關(guān)活動(dòng)和中時(shí)程互相關(guān)活動(dòng),長時(shí)程互相關(guān)活動(dòng)消缺失(n=20)。應(yīng)用100uM18p-GA可明顯降低CSNB大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的自發(fā)性放電頻率,而其對(duì)野生型大鼠視網(wǎng)膜的放電頻率無明顯影響(n-5)。 7. CSNB大鼠視網(wǎng)膜視覺信號(hào)傳導(dǎo)通路的重塑 按照單個(gè)神經(jīng)節(jié)細(xì)胞ON-型放電反應(yīng)頻率占其光誘導(dǎo)放電反應(yīng)總頻率的比例,將其歸為三類：1)ON反應(yīng)主導(dǎo)型神經(jīng)節(jié)細(xì)胞(比率0.5)；2)OFF反應(yīng)主導(dǎo)型神經(jīng)節(jié)細(xì)胞(比率0.5)；3)均衡反應(yīng)型神經(jīng)節(jié)細(xì)胞(比率=0.5)。較野生型大鼠,CSNB大鼠視網(wǎng)膜中的OFF反應(yīng)主導(dǎo)型神經(jīng)節(jié)細(xì)胞數(shù)占光誘導(dǎo)放電神經(jīng)節(jié)細(xì)胞總數(shù)的比例增加(Wt:60.6%; CSNB:78.1%),而ON反應(yīng)主導(dǎo)型神經(jīng)節(jié)細(xì)胞數(shù)所占的比例減小(Wt:31.5%; CSNB:18.2%)。對(duì)照兩種大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞光誘導(dǎo)ON-型和OFF-型反應(yīng)的潛伏期發(fā)現(xiàn)：CSNB大鼠的ON-型和OFF-型反應(yīng)潛伏期均較野生型大鼠顯著延長(P0.05)。應(yīng)用用CNQX+PDA可明顯阻斷CSNB大鼠和野生型大鼠視網(wǎng)膜光誘導(dǎo)OFF-型反應(yīng)：在兩種視網(wǎng)膜中,撤光型神經(jīng)節(jié)細(xì)胞(OFF RGCs)和給光-撤光型神經(jīng)節(jié)細(xì)胞(ON-OFF RGCs)的OFF反應(yīng)均可被阻斷劑消除,而延遲撤光型神經(jīng)節(jié)細(xì)胞(delayed OFF RGCs)和持續(xù)給光-撤光型神經(jīng)節(jié)細(xì)胞的OFF反應(yīng)無顯著變化。施加藥物雞尾酒后,在CSNB大鼠視網(wǎng)膜中,光誘導(dǎo)神經(jīng)節(jié)細(xì)胞放電反應(yīng)的平均頻率降低而ON-型反應(yīng)平均頻率增加(P0.01),且ON-型反應(yīng)的潛伏期顯著減小(P0.05)；Wt大鼠視網(wǎng)膜中的光誘導(dǎo)神經(jīng)節(jié)細(xì)胞放電反應(yīng)的平均頻率降低,但ON-型反應(yīng)的平均頻率和潛伏期無顯著變化(P0.05)。 結(jié)論 1.在視網(wǎng)膜光誘發(fā)反應(yīng)場(chǎng)電位記錄中,CSNB大鼠視網(wǎng)膜光誘發(fā)場(chǎng)電位仍可記錄到a-波,盡管其峰值顯著減小,說明CSNB大鼠光感受器細(xì)胞至少保存了部分活性和功能；其光誘發(fā)場(chǎng)電位波形中b-波缺失表明CSNB大鼠視網(wǎng)膜視覺信號(hào)傳導(dǎo)在第一級(jí)神經(jīng)元與第二級(jí)神經(jīng)元之間發(fā)生嚴(yán)重障礙。 2.在電刺激放電反應(yīng)記錄中,CSNB大鼠視網(wǎng)膜的長潛伏期反應(yīng)的放電頻率較野生型大鼠降低,表明Cacnalf基因突變后來源于視網(wǎng)膜外層的間接突觸刺激驅(qū)動(dòng)減少。 3. CSNB大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞自發(fā)放電的頻率升高表明：雖然CSNB大鼠視網(wǎng)膜外層視覺信號(hào)傳導(dǎo)發(fā)生障礙且光感受器細(xì)胞功能存在缺陷,但其視網(wǎng)膜內(nèi)層的自發(fā)性電生理活動(dòng)亢進(jìn)。 4. CSNB大鼠視網(wǎng)膜混合型放電細(xì)胞占所有自發(fā)放電神經(jīng)節(jié)細(xì)胞的比重升高,且混合型放電細(xì)胞的放電總數(shù)顯著增加,因此可以判定活躍的混合型放電細(xì)胞是CSNB大鼠視網(wǎng)膜自發(fā)性電生理活動(dòng)亢進(jìn)的主要原因。 5.信號(hào)/噪聲比下降表明CSNB大鼠視網(wǎng)膜分離有效視覺信號(hào)與背景噪聲的反應(yīng)能力降低,其神經(jīng)節(jié)細(xì)胞處理編碼視覺信號(hào)的有效度和可信度發(fā)生缺陷,且直接影響到視網(wǎng)膜視覺信號(hào)傳通路的效率。 6. CSNB大鼠視網(wǎng)膜記錄數(shù)據(jù)中長時(shí)程互相關(guān)活動(dòng)消缺失,可能是其視網(wǎng)膜L-型鈣通道失功能導(dǎo)致化學(xué)遞質(zhì)釋放障礙導(dǎo)致視網(wǎng)膜外層的多突觸環(huán)路信息輸入減少或消失所造成。隙連接在CSNB大鼠視網(wǎng)膜中神經(jīng)元協(xié)同放電活動(dòng)中發(fā)揮重要作用,且其表達(dá)水平或活性功能發(fā)生上調(diào)。 7. CSNB大鼠視網(wǎng)膜光感受器細(xì)胞和ON-型雙極細(xì)胞發(fā)生視覺信號(hào)傳導(dǎo)障礙后,其下游的視覺信號(hào)傳導(dǎo)通路確實(shí)發(fā)生了重塑：光誘導(dǎo)放電反應(yīng)頻率降低主要?dú)w因于其ON-型放電反應(yīng)減弱,OFF通路在內(nèi)層視網(wǎng)膜視覺信號(hào)傳導(dǎo)中占據(jù)主導(dǎo)地位,而ON通路與神經(jīng)節(jié)細(xì)胞之間發(fā)生了某種程度的分離和障礙。 綜上所述,CSNB大鼠視網(wǎng)膜外層神經(jīng)遞質(zhì)釋放障礙與內(nèi)層視覺信號(hào)傳導(dǎo)通路重塑之間聯(lián)系緊密,明確其視覺信號(hào)傳導(dǎo)通路重塑的機(jī)制和時(shí)機(jī)可為下一步制訂視覺保護(hù)干預(yù)措施、改進(jìn)完善治療方案提供依據(jù)。
[Abstract]:The congenital stationary night blindness ( CSNB ) is a non - progressive , inherited retinal disease based on visual signal system dysfunction .

The morphological changes in the outer layer of the retina are likely to affect the conduction integration function of the retinal inner layer . However , the morphological changes in the outer layer of the retina can affect the conduction integration function of the retinal inner layer . However , the morphological changes in the outer layer of the retina and the pathological effects on the ganglion cells in the innermost layer of the retina can be influenced by the morphological changes of the outer layer of the retina .

The multi - electrode array system has been widely used in the electrophysiological activity recording of retinal neurons . Because of its integrity and non - invasive advantage , the electrophysiological experiment has been applied to the physiological activities of retinal neurons such as photoreceptor cells and ganglion cells .

Materials and Methods

In the experiment of pharmacology , the electric signals collected by the electrodes of the multi - electrode array system are filtered and amplified by the lens system , and then applied to the perfusion environment in the bio - recording chip . The electric signals collected by the electrodes of the computer stimulator are filtered and amplified by the lens system .
In the electrical stimulation experiment , the electrophysiologic response of the retinal neurons was induced by the double - order equilibrium current stimulation sequence generated by the electrode 28 at the center of the array , and the recordings were collected for the other electrodes in the array and analyzed by the same software .

Results

1 . Retina induced field potential of wild - type SD rats and CSNB rats

The peak of a - wave was 0.196 鹵 0.038mv ( n - 20 ) . The peak of a - wave was significantly decreased ( 0.078 鹵 0.026mv , n - 20 , P0.01 ) , b - wave was absent .

2 . Electrical stimulation of electroretinogram in rats with wild - type SD rats and CSNB rats

Two types of electrical stimulation responses were found in the original waveform following the electrical stimulation of wild - type rats and CSNB rats following the application of the double - order equilibrium current stimulation : short latency response and long latency response . They were divided into 14 groups according to the distance between ganglion cells and current stimulation electrodes : ( 100 渭m , 14渭m , 200渭m , 22渭m , 36渭m , 38渭m , 400渭m , 424pm , 447 渭m , 500 渭m and 566 渭m ) . The results showed that the electrical stimulation response intensity of ganglion cells decreased with increasing distance from the current stimulating electrodes located in the center of the array ;
In the group , the number of discharges in the long incubation period of the retina of the CSNB rats was reduced compared with that of the wild type rats ( P0.01 ) .

3 . Spontaneous discharge of retinal spontaneous discharges of wild - type SD rats and CSNB rats

It was found that the spontaneous discharge frequency ( 9.93 鹵 4.14Hz , n = 386 ) of retinal ganglion cells in CSNB rats was significantly higher than that of wild type rats ( 2.610 . 73Hz , n = 295 , P0.01 ) . From the cumulative spontaneous discharge frequency graph , the spontaneous discharge frequency curve of the retinal ganglion cells of the CSNB rats was observed to be lower and shifted to the right compared with the wild - type rats .

4 . Type of spontaneous discharges of retinal ganglion cells from wild - type SD rats and CSNB rats

There were four types of ganglion cells producing spontaneous discharge reaction : burst type discharge cells , conventional type discharge cells , non - conventional type discharge cells and mixed type discharge cells . The proportion of the retinal burst type discharge cells and mixed type discharge cells in the CSNB rat was higher than that of the wild type rats ( Wt : explosion type 15.2 % , mixed type : 24.7 % , n - 20 ;
Compared with wild type rats ( Wt : 51.4 % , CSNB : 36.8 % , P0.01 ) . The total number of discharge cells ( 246,53 , n - 20 ) in the retinal ganglion cells of CSNB rats was significantly higher than that of the hybrid discharge cells ( 112089 , n = 20 , P0.01 ) . There was no significant difference in the total number of discharges in the retinal ganglion cells of CSNB rats and Wt rat retina ( P0.05 ) .

5 . Photoinduced discharges of retinal light induced by wild - type SD rats and CSNB rats

The retinal ganglion cells from CSNB rats and Wt rat retina can be divided into six categories : 1 ) to light type ( ON - type ) according to the discharge characteristics of the light reaction ;
2 ) Withdrawal type ( OFF - type ) ;
3 ) A - OFF type ( ON - OFF type ) ;
4 ) sustained - to - light type ( sustained ON type ) ;
5 ) sustained on - off type ( sustained ON - OFF type ) ;
6 ) Delayed OFF type . The average frequency ( 16.40 鹵 9.97Hz , n - 340 ) of retinal ganglion cells in CSNB rats was significantly lower than that of wild - type rats ( 38.6211 . 78Hz , n = 373 , P0.01 ) . Compared with the retina of Wt rat , the frequency of spontaneous discharge ( background noise ) in the retina of CSNB rats is high and the frequency of light - induced discharge reaction ( visual signal ) is low , so its signal / noise ratio ( S - N ration ) is reduced .

6 . Cross - correlation of spontaneous discharges between wild - type SD rats and CSNB rats

In the retina of wild type rats , three kinds of characteristic ganglia cell cross - correlation activities were observed : short - term cross - correlation activities , medium - duration cross - correlation activities and long - range cross - correlation activities . In the CSNB rat retinal record data only short - term cross - correlation and medium - time cross - correlation were observed , and the long - range cross - correlation activities were eliminated ( n = 20 ) .

7 . Reconstruction of retinal visual signal transduction pathways in CSNB rats

According to the ratio of the frequency of ON - type discharge reaction of individual ganglion cells to the total frequency of light - induced discharge reaction , they were classified into three groups : 1 ) ON reaction - dominant ganglion cells ( ratio 0.5 ) ;
2 ) OFF reaction leading ganglion cells ( ratio 0.5 ) ;
3 ) Compared with wild type rats , the proportion of the dominant ganglion cells in the retina of the CSNB rat was increased ( Wt : 60.6 % ; CSNB : 78.1 % ) , while the proportion of the dominant ganglion cells in the ON reaction was decreased ( Wt : 31.5 % ; CSNB : 18.2 % ) . The latency of the light - induced ON - type and OFF - type responses in both rat retinal ganglion cells was significantly prolonged compared with that of wild - type rats ( P0.05 ) .
The average frequency of light - induced ganglion cell discharge in the retina of Wt rat was decreased , but the average frequency and latency of the ON - type reaction did not change significantly ( P0.05 ) .

Conclusion

1 . In the field potential recording of retinal light - induced reaction , the retinal light - induced field potential of CSNB rats could still be recorded to a - wave , although its peak decreased significantly , indicating that CSNB rat photoreceptor cells had at least partial activity and function ;
The absence of b - wave in the light - induced field potential waveform indicates that the retinal visual signal transduction of CSNB rats is a serious obstacle between the first - level neurons and the second - level neurons .

2 . In the recording of electrical stimulation , the response to long latency of the retina of CSNB rats was lower than that of wild type rats , suggesting that the indirect synaptic stimulation of the outer retina of CSNB gene was decreased after gene mutation .

3 . The frequency of spontaneous discharge of retinal ganglion cells in CSNB rats showed that although the visual signal transduction of the retinal outer layer of CSNB rats was impaired and the function of photoreceptor cells was defective , the spontaneous electrophysiological activity of the retina inner layer was too high .

4 . CSNB rat retinal hybrid discharge cells accounted for a significant increase in the specific gravity of all spontaneous discharge ganglion cells , and the total number of discharge cells in the hybrid discharge cells was significantly increased , so it was possible to determine that the active hybrid discharge cells were the main reason for the spontaneous electrophysiologic activity of the retina of CSNB rats .

5 . The reduction of signal / noise ratio indicates that the response ability of retinal detachment effective visual signal and background noise in CSNB rats is reduced , and the effective degree and credibility of retinal ganglion cell processing coded visual signal are defective , and the efficiency of retinal visual signal transmission path is directly affected .

6 . The deletion of long - range cross - correlation activities in the retinal record data of CSNB rats may be caused by the loss of the retinal L - type calcium channel , which leads to the reduction or disappearance of the multi - synaptic ring information input in the outer layer of the retina .

7 . After the visual signal transduction of the retinal photoreceptor cells and the ON - type bipolar cells in the CSNB rats , the visual signal transduction pathways downstream of the cells were remolded : the decrease in the frequency of the photoinduced discharge was mainly attributed to the decrease of the ON - type discharge reaction , the OFF pathway dominated the visual signal transduction of the inner retina , while the ON pathway and the ganglion cells had some degree of separation and disorder .

In conclusion , the mechanism and timing of visual signal transduction pathway remodeling in the retinal outer layer of CSNB rats are closely related to the reconstruction of the inner layer visual signal transduction pathway , and the mechanism and timing of the reconstruction of the visual signal transduction pathway can be defined as the basis for the next step of developing visual protection interventions and improving the treatment scheme .
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R85

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