本文選題：M型乙酰膽堿受體 + 前庭終器��； 參考：《華中科技大學(xué)》2011年博士論文

【摘要】：第一部分M型乙酰膽堿受體蛋白在豚鼠前庭中的表達的實驗研究 [目的]mAChRs在中樞和外周神經(jīng)系統(tǒng)中廣泛表達,并在調(diào)節(jié)細胞活動和功能中起著重要的作用。乙酰膽堿(acetylcholine, ACh)是哺乳動物主要的內(nèi)耳傳出神經(jīng)遞質(zhì),在外周前庭系統(tǒng)中存在各種毒菌堿型乙酰膽堿受體(muscarinic acetylcholine receptor, mAChR)和煙堿型乙酰膽堿受體(nicotinic acetylcholine receptor, nAChR)。目前的研究表明M1-M5型乙酰膽堿受體在哺乳動物前庭終器中均有表達,但這些受體蛋白在豚鼠前庭中的表達尚無報道,本實驗研究編碼M型乙酰膽堿受體(muscarinic acetylcholine receptor, mAChR)的蛋白在豚鼠前庭終器中的表達。 [方法]從豚鼠內(nèi)耳中分離出前庭組織分別提取半規(guī)管(水平半規(guī)管,垂直半規(guī)管,后半規(guī)管),球囊,橢圓囊蛋白,應(yīng)用western blot法檢測M1-M5 mAChR蛋白在以上前庭終器中的表達。 [結(jié)果]在豚鼠前庭終器(半規(guī)管,橢圓囊,球囊)中均可檢測到M1-M5 mAChR蛋白的表達。 [結(jié)論]本研究運用western blot的方法成功的檢測出M1-M5 mAChR蛋白在豚鼠前庭終器(半規(guī)管,橢圓囊,球囊)中的表達,由此為M1-M5 mAChR在豚鼠前庭中的存在提供證據(jù),并證實M1-M5 mAChR也可能在介導(dǎo)前庭毛細胞膽堿能傳出過程中起到重要的作用,進而為前庭疾病機制闡明及前庭疾病的治療提供理論基礎(chǔ)。 第二部分M型乙酰膽堿受體在豚鼠前庭終器中的表達的實驗研究 [目的]目前研究證實乙酰膽堿(acetylcholine, ACh)在外周前庭系統(tǒng)中廣泛表達,在哺乳動物內(nèi)耳中存在各種毒菌堿型乙酰膽堿受體(muscarinic acetylcholine receptor, mAChR)和煙堿型乙酰膽堿受體(nicotinic acetylcholine receptor, nAChR),是哺乳動物內(nèi)耳的主要傳出神經(jīng)遞質(zhì)。目前的研究表明M1-M5型乙酰膽堿受體在哺乳動物前庭終器中均有表達,但這些受體在豚鼠前庭組織中的表達尚無報道,本實驗研究編碼M1-M5型乙酰膽堿受體(muscarinic acetylcholine receptor, mAChR)的mRNA在豚鼠前庭終器(半規(guī)管,橢圓囊,球囊)中的表達。 [方法]從豚鼠內(nèi)耳中分離出前庭組織并提取RNA,采用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(reverse transcription polymerase chain reaction, RT-PCR)法檢測編碼M1-M5mAChR的mRNA是否在豚鼠前庭組織中有表達。 [結(jié)果]在豚鼠前庭終器中可檢測到編碼M1-M5 mAChR基因的mRNA的表達,經(jīng)測序證實其與預(yù)計擴增的cDNA序列一致。 [結(jié)論]本研究運用RT-PCR的方法成功的檢測出編碼M1-M5 mAChR基因的mRNA在豚鼠前庭終器中的表達,由此為M1-M5 mAChR在豚鼠VHCsⅡ中的存在提供證據(jù),并證實M1-M5 mAChR也可能在介導(dǎo)前庭毛細胞膽堿能傳出過程中起到重要的作用,進而為前庭疾病機制闡明及前庭疾病的治療提供理論基礎(chǔ)。 第三部分m型乙酰膽堿受體在豚鼠前庭毛細胞中的表達的實驗研究 [目的]目前研究證實乙酰膽堿(acetylcholine, ACh)在外周前庭系統(tǒng)中廣泛表達,在哺乳動物內(nèi)耳中存在各種毒菌堿型乙酰膽堿受體(muscarinic acetylcholine receptor, mAChR)和煙堿型乙酰膽堿受體(nicotinic acetylcholine receptor, nAChR),是哺乳動物內(nèi)耳的主要傳出神經(jīng)遞質(zhì)。目前的研究表明M1-M5型乙酰膽堿受體在哺乳動物前庭終器中均有表達,但這些受體在豚鼠前庭毛細胞中的表達國內(nèi)外尚無報道,本實驗研究編碼M1-M5型乙酰膽堿受體(M1-M5 muscarinic acetylcholine receptor, M1-M5 mAChR)的mRNA在豚鼠前庭終器及Ⅱ型前庭毛細胞(vestibular hair cell, VHC)中的表達。 [方法]從豚鼠前庭組織中分離出Ⅱ型前庭毛細胞,在顯微操作儀器下應(yīng)用玻璃電極捕獲單個Ⅱ型前庭毛細胞,應(yīng)用單細胞RT-PCR(single cell reverse transcription polymerase chain reaction, SC-RT-PCR)法檢測編碼M1-M5 mAChR的mRNA是否在豚鼠Ⅱ型前庭毛細胞中有表達,從而明確M1-M5 mAChR在前庭組織中的細胞定位。 [結(jié)果]編碼M1-M5 mAChR基因的mRNA在豚鼠Ⅱ型前庭毛細胞中均可被檢測到,經(jīng)測序證實其與預(yù)計擴增的cDNA序列一致。 [結(jié)論]本研究運用單細胞RT-PCR的方法檢測編碼M1-M5 mAChR的mRNA在豚鼠VHCsⅡ中的表達,由此為M1-M5 mAChR在豚鼠VHCsⅡ中的存在提供證據(jù),并證實M1-M5mAChR也可能在介導(dǎo)前庭毛細胞膽堿能傳出過程中起到重要的作用,進而為前庭疾病機制闡明及前庭疾病的治療提供理論基礎(chǔ)。
[Abstract]:Part one: Experimental Study on the expression of M acetylcholine receptor protein in the vestibule of guinea pigs
[Objective]mAChRs is widely expressed in the central and peripheral nervous systems and plays an important role in regulating cell activity and function. Acetylcholine (ACh) is the main mammalian inner ear efferent neurotransmitter, and there are various virulent alkali acetylcholine receptors (muscarinic acetylcholine receptor) in the peripheral vestibule system. MAChR) and nicotinic acetylcholine receptor (nicotinic acetylcholine receptor, nAChR). Current studies have shown that M1-M5 type acetylcholine receptors are expressed in mammalian vestibule terminals, but the expression of these receptor proteins in the vestibule of guinea pigs is not reported. This experiment studies the encoding of M acetylcholine receptor (muscarinic acetylcholine). The expression of receptor, mAChR protein in the vestibular end organ of guinea pigs.
[Methods] the semicircular canals (horizontal semicircular canals, vertical semicircular canals, posterior semicircular canals), balloon and oval cysts were isolated from the inner ear of guinea pigs and detected the expression of M1-M5 mAChR protein in the above vestibular end organs by Western blot method.
[results] the expression of M1-M5 mAChR protein could be detected in the vestibular terminal organs (semicircular canal, utricle and balloon) of guinea pigs.
[Conclusion] this study successfully detected the expression of M1-M5 mAChR protein in the vestibular terminal (semicircular canals, oval sac and balloon) of guinea pigs by Western blot, which provided evidence for the existence of M1-M5 mAChR in the vestibule of guinea pigs, and confirmed that M1-M5 mAChR may also play an important role in mediating the cholinergic efferent of the vestibule hair cells. This will provide a theoretical basis for elucidating the mechanism of vestibular diseases and the treatment of vestibular diseases.
The second part is the experimental study of the expression of M acetylcholine receptor in the vestibular end organ of guinea pigs.
[Objective] the present study confirms that acetylcholine (ACh) is widely expressed in the peripheral vestibule system. There are various kinds of muscarinic acetylcholine receptor (mAChR) and nicotinic acetylcholine receptor (nicotinic acetylcholine receptor, nAChR) in mammalian inner ear, which are the inner ear of mammalian. The present study showed that the M1-M5 acetylcholine receptor was expressed in the mammalian vestibular end organ, but the expression of these receptors in the vestibule tissues of the guinea pig was not reported. This experiment studied the mRNA of the M1-M5 type acetylcholine receptor (muscarinic acetylcholine receptor, mAChR) in the vestibule end organ of the guinea pig. The expression in the tube, the oval sac, the balloon.
[Methods] vestibule tissue was isolated from the inner ear of guinea pig and RNA was extracted, and the expression of mRNA in the vestibule tissues of guinea pig was detected by reverse transcription polymerase chain reaction (reverse transcription polymerase chain reaction, RT-PCR).
[results] the expression of mRNA encoding M1-M5 mAChR gene could be detected in the vestibular end organ of guinea pigs, and it was confirmed by sequencing that it was consistent with the predicted cDNA sequence.
[Conclusion] this study successfully detected the expression of mRNA encoding M1-M5 mAChR gene in the vestibular end organ of guinea pigs by using RT-PCR method, which provided evidence for the existence of M1-M5 mAChR in the guinea pig VHCs II, and confirmed that M1-M5 mAChR may also play an important role in mediating the cholinergic efferent of vestibular hair cells, and then the vestibular disease is a vestibular disease. Elucidating the pathogenesis and providing a theoretical basis for the treatment of vestibular diseases.
The third part is the experimental study of the expression of M acetylcholine receptor in guinea pig vestibular hair cells.
[Objective] the present study confirms that acetylcholine (ACh) is widely expressed in the peripheral vestibule system. There are various kinds of muscarinic acetylcholine receptor (mAChR) and nicotinic acetylcholine receptor (nicotinic acetylcholine receptor, nAChR) in mammalian inner ear, which are the inner ear of mammalian. The present study shows that the M1-M5 acetylcholine receptor is expressed in the mammalian vestibular end organ, but the expression of these receptors in the vestibular hair cells of the guinea pig is not yet reported. This experiment studies the mRNA of the M1-M5 muscarinic acetylcholine receptor (M1-M5 mAChR) encoding the M1-M5 type acetylcholine receptor in this experiment. Expression of vestibular and hair cell (VHC) in the vestibular end organ of guinea pigs.
[Methods] the type II vestibular hair cells were isolated from the vestibular tissue of guinea pigs, and the single type II vestibular hair cells were captured with glass electrodes under the micromanipulation instrument. The single cell RT-PCR (single cell reverse transcription polymerase chain reaction, SC-RT-PCR) method was used to detect whether the mRNA in the vestibule of M1-M5 mAChR was in the vestibular capillaries of the guinea pig type II. The expression of M1-M5 mAChR in the vestibular tissue is clear.
[results] mRNA encoding M1-M5 mAChR gene could be detected in guinea pig type II vestibular hair cells, and it was confirmed by sequencing that it was consistent with the predicted cDNA sequence.
[Conclusion] the present study used the single cell RT-PCR method to detect the expression of the mRNA encoding M1-M5 mAChR in the guinea pig VHCs II, thus providing evidence for the existence of M1-M5 mAChR in the guinea pig VHCs II, and confirmed that M1-M5mAChR may also play an important role in mediating the cholinergic efferent of vestibular hair cells, and further elucidate the mechanism of vestibular disease. It provides a theoretical basis for the treatment of vestibule disease.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2011
【分類號】：R764

【參考文獻】

相關(guān)期刊論文 前1條

1 ;Expression of nicotinic acetylcholine receptor subunit α9 in type Ⅱ vesti-bular hair cells of rats[J];Acta Pharmacologica Sinica;2006年11期

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本文編號：1914243