小鼠胚胎发育时期脑内SP和SPR的定位
发布时间：2018-08-05 13:17
【摘要】：【背景和目的】 P物质(SP)广泛分布于中枢神经系统和外周神经系统中,是由中枢和外周初级传入神经末梢释放的一种神经递质。哺乳动物的SP由前速激肽原(Preprotachykinin,PPT)A基因编码。PPT—A基因同样编码其他速激肽如neurokinin A,neuropeptide B等。速激肽受体也被分为速激肽NK_1受体,速激肽NK_2受体,速激肽NK_3受体。速激肽NK_1受体与SP有着最强亲和力,即P物质受体(SPR)。SP与SPR特异结合后使受体内化,通过激活磷脂酶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)两种第二信使发挥生物学效应。 已有的实验发现在小鼠胚胎时期脑内已有SP表达,但其与中枢神经系统发育的关系仍不清楚,而且关于SPR在小鼠胚胎时期脑发育过程中的表达也未见报道,因此调查SP与SPR在小鼠胚胎时期脑的分布情况及其相互间关系,可以为进一步研究SP在中枢神经系统发育阶段所起的作用提供形态学依据。本研究采用免疫组织化学染色方法结合western blotting半定量方法,使用特异性SP和SPR抗体,观察SP和SPR在小鼠胚胎发育时期脑内表达的变化,推测胚胎发育阶段SP在中枢神经系统中所起的作用。 【材料与方法】 1.实验动物及组织切片制备 分别取E11(胚胎11天),E13,E15,E17,E19和P0(新生)ICR品系小鼠脑经冠状切面进行连续冰冻切片。 2.酶标免疫组织化学 将冰冻切片分别滴加兔抗SP多克隆抗体和抗SPR多克隆抗体,生物素标记的羊抗兔IgG,加入抗生物素蛋白—辣根过氧化物酶复合物,二甲基联苯胺呈色观察。
[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.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R321
本文编号：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.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R321
【参考文献】
相关期刊论文 前1条
1 包新民,舒斯云,包蓉,张魁华;P物质和受体在纹状体边缘区内的分布及其与学习记忆功能的关系[J];第一军医大学学报;2002年02期
，本文编号：2165911
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2165911.html
最近更新
教材专著
