本文選題：腎臟 + 病毒示蹤技術(shù)��； 參考：《華中科技大學(xué)》2012年博士論文

【摘要】：近年來(lái)隨著有生物活性的能夠進(jìn)行多突觸傳遞的嗜神經(jīng)示蹤病毒的發(fā)現(xiàn)、改造與應(yīng)用,有關(guān)調(diào)控內(nèi)臟功能的中樞核團(tuán)定位和功能認(rèn)識(shí)得以深化,相關(guān)中樞核團(tuán)之間的神經(jīng)解剖學(xué)通路變得更加明晰。本研究運(yùn)用偽狂犬病毒(pseudorabies virus, PRV)轉(zhuǎn)突觸示蹤技術(shù)和熒光標(biāo)記技術(shù)試圖確定中樞神經(jīng)系統(tǒng)參與調(diào)控小鼠腎臟功能的相關(guān)神經(jīng)核團(tuán),以及這些核團(tuán)之間的神經(jīng)解剖學(xué)投射通路。研究方法與結(jié)果方法：將嗜神經(jīng)示蹤病毒PRV-614注入19只成年雄性C57BL/6J小鼠的左側(cè)腎臟上極,按照動(dòng)物的不同生存期限無(wú)痛處死后制備冰凍切片,用熒光免疫組織化學(xué)技術(shù)檢測(cè)脊髓、腦干(延髓和中腦)中PRV-614感染神經(jīng)元、色氨酸羥化酶(TPH)和酪氨酸羥化酶(TH)陽(yáng)性神經(jīng)元的表達(dá)。結(jié)果：1.PRV-614經(jīng)小鼠腎臟注射后第3d,少量的PRV-614感染神經(jīng)元逆行示蹤標(biāo)記在脊髓T9的IML區(qū)。2.注射PRV-614后第4d,少量的PRV-614感染神經(jīng)元發(fā)現(xiàn)在脊髓T8-10的IML區(qū)、IC區(qū)和CAN區(qū),以及延髓頭端腹外側(cè)核。3.注射PRV-614后第5d,較多的PRV-614感染神經(jīng)元發(fā)現(xiàn)在脊髓T7-12的IML區(qū)、IC區(qū)和CAN區(qū)等,延髓頭端腹內(nèi)側(cè)核、延髓頭端腹外側(cè)核、延髓中縫核、孤束核、A5區(qū)域、藍(lán)斑核及亞藍(lán)斑核等；較少的PRV-614感染神經(jīng)元發(fā)現(xiàn)在中腦PPTg和LDTg區(qū)等。4.注射PRV-614后第6d,在脊髓幾乎未發(fā)現(xiàn)PRV-614感染神經(jīng)元。較多的PRV-614感染神經(jīng)元發(fā)現(xiàn)在延髓頭端腹內(nèi)側(cè)核、延髓頭端腹外側(cè)核、延髓中縫核、A5區(qū)域、藍(lán)斑核及亞藍(lán)斑核等,以及中腦PPTg和LDTg區(qū)。其中PRV-614感染神經(jīng)元主要發(fā)現(xiàn)在LDTg頭端和中間部位,以及PPTg中部和尾部。PRV-614/TPH雙重標(biāo)記神經(jīng)元主要分布在LDTg頭端,而PRV-614/TH神經(jīng)元分散在LDTg中在3個(gè)部位。PRV-614/TPH和PRV-614/TH神經(jīng)元主要位于PPTg (cPPTg)尾側(cè)。研究總結(jié)1.從小鼠腎臟注射的PRV-614經(jīng)突觸逆行傳遞到達(dá)中樞核團(tuán)具有時(shí)空表達(dá)特異性。2.主要調(diào)節(jié)腎臟交感神經(jīng)活性的脊髓節(jié)段范圍可能是T8-10,其中T9尤為重要。3.延髓頭端腹外側(cè)核是主要調(diào)節(jié)腎臟TH陽(yáng)性交感神經(jīng)活性的中樞核團(tuán),而延髓中縫核是主要調(diào)節(jié)腎臟TPH陽(yáng)性交感神經(jīng)活性的中樞核團(tuán)。4.腦橋中A5區(qū)域、藍(lán)斑核及亞藍(lán)斑核是主要調(diào)節(jié)腎臟TH陽(yáng)性交感神經(jīng)活性的中樞核團(tuán)。5. LDTg頭部和PPTg尾部均參與腎臟TH/TPH陽(yáng)性交感神經(jīng)活性的調(diào)節(jié)。
[Abstract]:In recent years, with the discovery, modification and application of bioactive eosinophilic tracer virus capable of multi-synaptic transmission, the localization and functional understanding of central nuclei regulating visceral function have been deepened. The neuroanatomic pathways between the relevant central nuclei became clearer. In this study, pseudorabies virus (PRV) synaptic tracer technique and fluorescence labeling technique were used to identify the central nervous system involved in the regulation of renal function in mice, as well as the neuroanatomical projection pathway between these nuclei. Methods and results: a total of 19 adult male C57BL/6J mice were injected with eosinophilic tracer virus (PRV-614) into the upper pole of the left kidney. Frozen sections were prepared after painless death according to the different survival periods of the animals. The spinal cord was detected by fluorescence immunohistochemical technique. Expression of PRV-614 positive neurons, tryptophan hydroxylase and tyrosine hydroxylase in the brain stem (medulla oblongata and midbrain). Results 1. On the 3rd day after injection of PRV-614 into the kidney of mice, a small number of PRV-614 infected neurons were retrogradely labeled in the IML region of spinal cord T9. On the 4th day after PRV-614 injection, a small number of PRV-614 infected neurons were found in the IML and CAN regions of the T8-10 spinal cord, as well as in the ventrolateral nucleus ventrolateral of the medulla oblongata. On the 5th day after PRV-614 injection, more PRV-614 infected neurons were found in the IML and CAN regions of T7-12 spinal cord, the ventromedial nucleus of the medulla oblongata, the ventrolateral nucleus of the medulla oblongata, the nucleus raphe nucleus of the medulla oblongata, the A5 region of the nucleus tractus solitarius, the nucleus coeruleus and the nucleus coeruleus. Fewer PRV-614 infected neurons were found in the PPTg and LDTg regions of the midbrain. On the 6th day after PRV-614 injection, almost no PRV-614 infected neurons were found in the spinal cord. More PRV-614 infected neurons were found in the ventromedial nucleus of the medulla oblongata, the ventrolateral nucleus of the medulla oblongata, the A5 area of the nucleus raphe nucleus of the medulla oblongata, the nucleus coeruleus and the sublocus coeruleus, and the PPTg and LDTg areas of the midbrain. The PRV-614 infection neurons were mainly located in the head and middle part of LDTg, and in the middle and tail of PPTg. The double labeled neurons were mainly located in the LDTg head, while the PRV-614/TH neurons were scattered in the three sites of LDTg. The PRV-614/TH neurons were mainly located in the caudal side of PPTg cPPTg. Research summary 1. The PRV-614 injected from the kidney of mice reached the central nucleus via synaptic retrograde transmission with spatiotemporal expression specificity. 2. The range of spinal cord segments which mainly regulate renal sympathetic nerve activity may be T 8-10, of which T 9 is especially important. 3. The ventrolateral nucleus of the medulla oblongata is the central nucleus which mainly regulates the activity of the th positive sympathetic nerve in the kidney, while the nucleus raphe nucleus of the medulla oblongata is the central nucleus which mainly regulates the sympathetic activity of the kidney TPH positive. The A5 region of the pons, the nucleus coeruleus and the sublocus coeruleus nucleus are the central nuclei which mainly regulate the sympathetic activity of the renal th positive neurons. Both LDTg head and PPTg tail are involved in the regulation of renal TH/TPH positive sympathetic nerve activity.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R322

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