溫度敏感TRP離子通道的異源組裝和功能研究
發(fā)布時(shí)間：2018-12-13 19:35
【摘要】：溫度感覺作為人體的一種重要感覺系統(tǒng),對(duì)于感受周圍環(huán)境變化,躲避危險(xiǎn)性傷害以及維持機(jī)體內(nèi)環(huán)境的穩(wěn)定都具有非常重要的意義。長期以來對(duì)溫度感覺機(jī)理的研究一直停留于直觀描述階段。造成溫度感覺研究長期落后的主要原因在于溫度感受器分子一直沒有被確定。溫度敏感TRP(Transient Receptor Potential)離子通道被分子克隆并證實(shí)為主要的細(xì)胞溫度感受器,為在分子水平上研究溫度感覺的分子機(jī)制提供了實(shí)驗(yàn)基礎(chǔ)。 作者前期的研究工作表明溫度敏感TRPV通道亞基間可以交互雜合組裝為新的通道,雜合通道的類型和其不同的亞基配比有關(guān),分子組裝過程中亞基配比為隨機(jī)的,雜合通道的電導(dǎo)和門控動(dòng)力學(xué)特性介于純合母體通道之間。通道亞基間的異源組裝有助于離子通道在不同組織,不同細(xì)胞類型以及不同生理狀態(tài)下功能多樣性的廣泛擴(kuò)展。近年來對(duì)于溫度敏感TRP通道的異源組裝研究也有大量報(bào)道,但雜合通道的功能方面我們卻知之甚少。 在本課題主要研究了雜合TRPV1/TRPV3通道在熱、化合物及電位激活過程中的通道功能特性。同時(shí)作者還將通道異源組裝的研究擴(kuò)展到其它溫度敏感TRP通道蛋白亞基。結(jié)果表明雜合TRPV1/TRPV3通道對(duì)TRPV1的激活劑和拮抗劑也都非常敏感;雜合TRPV1/TRPV3通道具有特定的溫度敏感性、激活溫度閾值和由熱引發(fā)的通道敏化現(xiàn)象。雜合通道門控特性的改變顯然是由TRPV1和TRPV3通道亞基間的相互作用所造成的。溫度敏感TRP通道異源組裝的實(shí)驗(yàn)研究中,作者證明了在細(xì)胞中表達(dá)TRPV1-TRPV3的串聯(lián)體能夠形成單一的雜合TRPV1/TRPV3通道,通過內(nèi)面向外(inside-out)的膜片鉗記錄模式可以從表達(dá)串聯(lián)體的細(xì)胞中記錄到單通道電流。甚至單通道電導(dǎo)也很一致地處于純合TRPV1和TRPV3通道的電導(dǎo)之間。這個(gè)結(jié)果和作者前期發(fā)表的有關(guān)TRPV1和TRPV3共同表達(dá)于細(xì)胞中可以組成雜合通道的結(jié)果一致。而對(duì)其它溫度敏感TRP離子通道的異源組裝研究顯示其蛋白亞基間不能形成雜合通道。 綜上所述,本研究證明了雜合TRPV1/TRPV3通道在細(xì)胞中的異源組裝以及雜合通道在熱激活及化學(xué)激活過程中功能的擴(kuò)展,雜合通道的形成和功能特性的擴(kuò)展也許有助于機(jī)體更加精確地對(duì)各種感覺和痛覺的敏感性進(jìn)行細(xì)微的調(diào)節(jié),從而更好地保護(hù)機(jī)體躲避有害刺激。
[Abstract]:As an important sensory system of human body, temperature sense is of great significance to feel the changes of the surrounding environment, avoid dangerous damage and maintain the stability of the body's internal environment. For a long time, the research on temperature sensing mechanism has been in the stage of visual description. The main reason for the long lag in the study of temperature sensing is that the thermoreceptor molecules have not been identified. The thermo-sensitive TRP (Transient Receptor Potential) ion channel was cloned and confirmed as the main cellular thermoreceptor, which provides the experimental basis for studying the molecular mechanism of temperature sensing at the molecular level. The previous work of the authors showed that the cross-heterozygous subunits of temperature-sensitive TRPV channels could be assembled into new channels, and the types of heterozygotes were related to their different subunit ratios, and the ratio of subunits in the molecular assembly process was random. The conductivity and gated dynamics of hybrid channels are between homozygous parent channels. The heterologous assembly of channel subunits contributes to the extensive expansion of ion channels in different tissues, cell types and physiological states. In recent years, there have been a lot of reports on the heterologous assembly of temperature-sensitive TRP channels, but little is known about the function of heterozygotes. In this paper, the functional characteristics of heterozygous TRPV1/TRPV3 channels during heat, compound and potential activation were studied. At the same time, the study of channel heterologous assembly was extended to other temperature sensitive TRP channel protein subunits. The results showed that the heterozygous TRPV1/TRPV3 channels were also sensitive to the activators and antagonists of TRPV1, and the heterozygous TRPV1/TRPV3 channels had specific temperature sensitivity, activation threshold and heat-induced channel sensitization. The change of heterozygous channel gating is obviously caused by the interaction between TRPV1 and TRPV3 channel subunits. In an experimental study of the heterologous assembly of thermo-sensitive TRP channels, the authors have demonstrated that a single heterozygous TRPV1/TRPV3 channel can be formed by the cascade expressing TRPV1-TRPV3 in cells. A single channel current can be recorded from the cells expressing the tandem through the patch-clamp recording mode of the inner outward (inside-out). Even the single channel conductance is very consistent between the homozygous TRPV1 and the TRPV3 channel conductance. This result is consistent with the results previously published by the authors that TRPV1 and TRPV3 co-express in cells to form heterozygous channels. Heterologous assembly of other temperature-sensitive TRP ion channels showed that heterozygous channels could not be formed between their protein subunits. In conclusion, this study demonstrated the heterogenous assembly of heterozygous TRPV1/TRPV3 channels in cells and the expansion of heterozygous channels in the processes of thermal and chemical activation. The formation of heterozygous channels and the expansion of functional characteristics may help the body to regulate the sensitivity of various senses and pain sensations more accurately and thus better protect the body from harmful stimuli.
【學(xué)位授予單位】：大連理工大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R339.1;Q43
本文編號(hào)：2377095
[Abstract]:As an important sensory system of human body, temperature sense is of great significance to feel the changes of the surrounding environment, avoid dangerous damage and maintain the stability of the body's internal environment. For a long time, the research on temperature sensing mechanism has been in the stage of visual description. The main reason for the long lag in the study of temperature sensing is that the thermoreceptor molecules have not been identified. The thermo-sensitive TRP (Transient Receptor Potential) ion channel was cloned and confirmed as the main cellular thermoreceptor, which provides the experimental basis for studying the molecular mechanism of temperature sensing at the molecular level. The previous work of the authors showed that the cross-heterozygous subunits of temperature-sensitive TRPV channels could be assembled into new channels, and the types of heterozygotes were related to their different subunit ratios, and the ratio of subunits in the molecular assembly process was random. The conductivity and gated dynamics of hybrid channels are between homozygous parent channels. The heterologous assembly of channel subunits contributes to the extensive expansion of ion channels in different tissues, cell types and physiological states. In recent years, there have been a lot of reports on the heterologous assembly of temperature-sensitive TRP channels, but little is known about the function of heterozygotes. In this paper, the functional characteristics of heterozygous TRPV1/TRPV3 channels during heat, compound and potential activation were studied. At the same time, the study of channel heterologous assembly was extended to other temperature sensitive TRP channel protein subunits. The results showed that the heterozygous TRPV1/TRPV3 channels were also sensitive to the activators and antagonists of TRPV1, and the heterozygous TRPV1/TRPV3 channels had specific temperature sensitivity, activation threshold and heat-induced channel sensitization. The change of heterozygous channel gating is obviously caused by the interaction between TRPV1 and TRPV3 channel subunits. In an experimental study of the heterologous assembly of thermo-sensitive TRP channels, the authors have demonstrated that a single heterozygous TRPV1/TRPV3 channel can be formed by the cascade expressing TRPV1-TRPV3 in cells. A single channel current can be recorded from the cells expressing the tandem through the patch-clamp recording mode of the inner outward (inside-out). Even the single channel conductance is very consistent between the homozygous TRPV1 and the TRPV3 channel conductance. This result is consistent with the results previously published by the authors that TRPV1 and TRPV3 co-express in cells to form heterozygous channels. Heterologous assembly of other temperature-sensitive TRP ion channels showed that heterozygous channels could not be formed between their protein subunits. In conclusion, this study demonstrated the heterogenous assembly of heterozygous TRPV1/TRPV3 channels in cells and the expansion of heterozygous channels in the processes of thermal and chemical activation. The formation of heterozygous channels and the expansion of functional characteristics may help the body to regulate the sensitivity of various senses and pain sensations more accurately and thus better protect the body from harmful stimuli.
【學(xué)位授予單位】：大連理工大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R339.1;Q43
【引證文獻(xiàn)】
相關(guān)期刊論文 前1條
1 陳曉博;曹鵬;于鋒;王大為;;溫度敏感瞬時(shí)受體電位通道參與化療引起的外周神經(jīng)病變的研究進(jìn)展[J];藥學(xué)與臨床研究;2014年02期
，本文編號(hào)：2377095
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