小鼠胚胎發(fā)育時(shí)期腦內(nèi)SP和SPR的定位
發(fā)布時(shí)間：2018-08-05 13:17
【摘要】：【背景和目的】 P物質(zhì)(SP)廣泛分布于中樞神經(jīng)系統(tǒng)和外周神經(jīng)系統(tǒng)中,是由中樞和外周初級(jí)傳入神經(jīng)末梢釋放的一種神經(jīng)遞質(zhì)。哺乳動(dòng)物的SP由前速激肽原(Preprotachykinin,PPT)A基因編碼。PPT—A基因同樣編碼其他速激肽如neurokinin A,neuropeptide B等。速激肽受體也被分為速激肽NK_1受體,速激肽NK_2受體,速激肽NK_3受體。速激肽NK_1受體與SP有著最強(qiáng)親和力,即P物質(zhì)受體(SPR)。SP與SPR特異結(jié)合后使受體內(nèi)化,通過激活磷脂酶C(PLC)并水解磷酸肌醇(PIP_2) 生成三磷酸肌醇(IP_3)和二酯酰甘油(DAG)兩種第二信使發(fā)揮生物學(xué)效應(yīng)。 已有的實(shí)驗(yàn)發(fā)現(xiàn)在小鼠胚胎時(shí)期腦內(nèi)已有SP表達(dá),但其與中樞神經(jīng)系統(tǒng)發(fā)育的關(guān)系仍不清楚,而且關(guān)于SPR在小鼠胚胎時(shí)期腦發(fā)育過程中的表達(dá)也未見報(bào)道,因此調(diào)查SP與SPR在小鼠胚胎時(shí)期腦的分布情況及其相互間關(guān)系,可以為進(jìn)一步研究SP在中樞神經(jīng)系統(tǒng)發(fā)育階段所起的作用提供形態(tài)學(xué)依據(jù)。本研究采用免疫組織化學(xué)染色方法結(jié)合western blotting半定量方法,使用特異性SP和SPR抗體,觀察SP和SPR在小鼠胚胎發(fā)育時(shí)期腦內(nèi)表達(dá)的變化,推測(cè)胚胎發(fā)育階段SP在中樞神經(jīng)系統(tǒng)中所起的作用。 【材料與方法】 1.實(shí)驗(yàn)動(dòng)物及組織切片制備 分別取E11(胚胎11天),E13,E15,E17,E19和P0(新生)ICR品系小鼠腦經(jīng)冠狀切面進(jìn)行連續(xù)冰凍切片。 2.酶標(biāo)免疫組織化學(xué) 將冰凍切片分別滴加兔抗SP多克隆抗體和抗SPR多克隆抗體,生物素標(biāo)記的羊抗兔IgG,加入抗生物素蛋白—辣根過氧化物酶復(fù)合物,二甲基聯(lián)苯胺呈色觀察。
[Abstract]:[background and objective] substance P (SP), which is widely distributed in the central nervous system and peripheral nervous system, is a neurotransmitter released from the central and peripheral primary afferent nerve endings. Sp in mammals is encoded by preprotachykinin PPT A gene. PPT-A gene also encodes other fast kinins such as neurokinin Agnopeptide B. Tachykinin receptors are also divided into tachykinin NK_1 receptors, tachykinin NK_2 receptors and tachykinin NK_3 receptors. Tachykinin NK_1 receptor has the strongest affinity with SP, that is, substance P receptor (SPR). Sp binds specifically with SPR to make the receptor in vivo. By activating phospholipase C (PLC) and hydrolyzing inositol phosphate (PIP_2) to produce inositol triphosphate (IP_3) and diester glycerol (DAG), two second messengers play a biological role. However, the relationship between SP expression and central nervous system development is still unclear, and the expression of SPR in mouse embryonic brain has not been reported. Therefore, the investigation of the distribution of SP and SPR in mouse embryonic brain and the relationship between them can provide morphological basis for further study on the role of SP in the development of the central nervous system. In this study, immunohistochemical staining combined with western blotting semi-quantitative method was used to observe the expression of SP and SPR in mouse brain during embryonic development by using specific SP and SPR antibodies. We speculated the role of SP in the central nervous system during embryonic development. [materials and methods] 1. E11 (embryonic 11 days) and P0 (newborn) ICR strain E19 and P0 (newborn) ICR strain were prepared by coronal section respectively and frozen sections were performed on the brain of E11 (embryonic 11 days) and P0 (newborn) ICR mice respectively. 2. The frozen sections were added with rabbit anti-SP polyclonal antibody and anti-SPR polyclonal antibody, respectively. Biotin labeled goat anti rabbit IgG was added with biotin protein-horseradish peroxidase complex, and dimethyl benzidine was observed in color.
【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】：2006
【分類號(hào)】：R321
本文編號(hào)：2165911
[Abstract]:[background and objective] substance P (SP), which is widely distributed in the central nervous system and peripheral nervous system, is a neurotransmitter released from the central and peripheral primary afferent nerve endings. Sp in mammals is encoded by preprotachykinin PPT A gene. PPT-A gene also encodes other fast kinins such as neurokinin Agnopeptide B. Tachykinin receptors are also divided into tachykinin NK_1 receptors, tachykinin NK_2 receptors and tachykinin NK_3 receptors. Tachykinin NK_1 receptor has the strongest affinity with SP, that is, substance P receptor (SPR). Sp binds specifically with SPR to make the receptor in vivo. By activating phospholipase C (PLC) and hydrolyzing inositol phosphate (PIP_2) to produce inositol triphosphate (IP_3) and diester glycerol (DAG), two second messengers play a biological role. However, the relationship between SP expression and central nervous system development is still unclear, and the expression of SPR in mouse embryonic brain has not been reported. Therefore, the investigation of the distribution of SP and SPR in mouse embryonic brain and the relationship between them can provide morphological basis for further study on the role of SP in the development of the central nervous system. In this study, immunohistochemical staining combined with western blotting semi-quantitative method was used to observe the expression of SP and SPR in mouse brain during embryonic development by using specific SP and SPR antibodies. We speculated the role of SP in the central nervous system during embryonic development. [materials and methods] 1. E11 (embryonic 11 days) and P0 (newborn) ICR strain E19 and P0 (newborn) ICR strain were prepared by coronal section respectively and frozen sections were performed on the brain of E11 (embryonic 11 days) and P0 (newborn) ICR mice respectively. 2. The frozen sections were added with rabbit anti-SP polyclonal antibody and anti-SPR polyclonal antibody, respectively. Biotin labeled goat anti rabbit IgG was added with biotin protein-horseradish peroxidase complex, and dimethyl benzidine was observed in color.
【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】：2006
【分類號(hào)】：R321
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 包新民,舒斯云,包蓉,張魁華;P物質(zhì)和受體在紋狀體邊緣區(qū)內(nèi)的分布及其與學(xué)習(xí)記憶功能的關(guān)系[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2002年02期
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