NDRG2在大鼠前脑及人胚胎呼吸系统分布特点的研究

发布时间：2018-03-03 12:56

  本文选题：ndrg2　切入点：大鼠前脑　出处：《第四军医大学》2007年硕士论文　论文类型：学位论文

【摘要】： 【研究背景】 Ndrg2与ndrg1、ndrg3和ndrg4共同构成了一个新的基因家族,即ndrg家族。该基因家族参与了细胞的生长,但具体机制仍不清楚。人ndrg2由我的导师邓艳春教授发现,其基因组全长为9.33kb,位于染色体的14q11.2,mRNA全长2024bp,编码含357个氨基酸残基,分子量约为40.79kD,因此曾命名为p40和SYLD。同源性分析表明,其与ndrg1(也称做脑发育相关蛋白)具有高度同源性,并含有一个酰基携带蛋白结构域,故称为ndrg2。研究结果初步显示,ndrg2在多种正常组织中高表达,而在某些肿瘤组织和肿瘤细胞系中低表达或无表达,推测该基因可能与肿瘤的发生发展有关,因此提出ndrg2可能为新的抑癌候选基因。新近资料表明,ndrg2参与了抑郁症和阿尔茨海默氏病等脑部疾病的发生发展,NDRG2的表达还受肾上腺类固醇和抗抑郁药物的调节,进行该研究时还发现NDRG2广泛表达于星形胶质细胞。我室近期研究发现NDRG2蛋白在小鼠和人胚胎发育过程中具有一定的时间和空间表达特性,随着组织、器官的成熟表达更为显著和广泛。此外NDRG2蛋白在小鼠生后发育过程中心、脑、肝、肾和骨骼肌中具有一定时间和空间表达特性,NDRG2蛋白广泛表达于成年小鼠的多种组织。侯双兴等研究表明NDRG2在胎脑发育过程中具有一定时间空间的表达特性,认为NDRG2在胎脑各脑功能区中广泛表达,并随着脑发育的成熟,表达更为显著,从而推测ndrg2可能对神经细胞发育、成熟有重要影响,同ndrg1一样是一种脑发育相关基因,但其在出生后脑发育各阶段的表达情况还没有人研究;而且作为发育相关基因ndrg2在中枢神经系统外的其他系统发育过程的表达有什么规律,目前也不清楚。 【研究目的】: 1.探讨NDRG2在出生后不同年龄大鼠前脑的表达规律。 2.探讨NDRG2在不同胎龄16～28周人胚胎呼吸系统的表达情况。 【研究方法】: 1.免疫荧光双重标记法从蛋白水平确定NDRG2在不同年龄大鼠前脑的表达特点及细胞定位。 取1天(新生儿期)、60天(青年期)及240天(中年期)大鼠前脑组织,用NDRG2与GFAP免疫荧光双重标记法观察NDRG2蛋白在大鼠前脑的表达规律(数目、形态及分布)及NDRG2同GFAP的位置关系。 2.免疫组织化学法从蛋白水平确定NDRG2在16～28周人胚胎呼吸系统的表达特点及细胞定位。 搜集16～28周人工流产胎儿呼吸系统的肺和气管组织,制成石蜡切片,用抗NDRG2单克隆抗体,行免疫组化染色(ABC法),从蛋白水平观察NDRG2表达情况。统计阳性细胞数,利用统计学方法,判断不同胎龄的肺和气管间NDRG2表达有无差别。 【结果】: 1.可见NDRG2在苍白球、皮质、梨状皮质、背侧丘脑、下丘脑、脉络丛等部位有广泛的表达,免疫阳性产物在灰质中的表达多于白质。值得注意的是在1天及60天大鼠的前脑中,NDRG2蛋白主要表达于神经细胞、室管膜细胞及脉络丛上皮细胞的胞浆中,未观察到与GFAP共表达;而在240天的大鼠脑组织中,在苍白球、皮质、视神经、背侧丘脑及下丘脑可见NDRG2蛋白与GFAP阳性细胞共表达。此外本研究首次发现3个年龄段的大鼠视上核神经细胞内均可观察到NDRG2蛋白的表达。 2. NDRG2在16～28周胚胎呼吸系统中有广泛的表达,阳性产物主要位于上皮细胞的胞浆中,但是不同胎龄之间NDRG2的表达未见明显差别。 【结论】: 1. NDRG2蛋白在1天及60天大鼠前脑内的神经细胞广泛表达,可能是ndrg2作为发育相关基因,广泛参与中枢神经系统生长发育的结果。在240天大鼠前脑观察到NDRG2蛋白与GFAP共表达,而在1天及60天大鼠脑内未观察到NDRG2蛋白与GFAP共表达,提示中年大鼠前脑内星形胶质细胞也表达NDRG2。此外,本研究首次发现NDRG2蛋白在3个年龄段大鼠下丘脑视上核的神经细胞均有表达。 2. NDRG2蛋白在16～28周人胚胎呼吸系统的假复层柱状纤毛上皮和单层立方上皮有广泛的表达,提示ndrg2在中、末期胚胎的呼吸系统上皮细胞的生长与发育过程中起一定作用。NDRG2阳性产物定位于上皮细胞的胞浆,提示NDRG2可能是一种胞浆蛋白。
[Abstract]:[research background]
Ndrg2 and NDRG1, NDRG3 and NDRG4 constitute a new gene family, NDRG family. This family of genes involved in cell growth, but the mechanism is not clear. NDRG2 by my tutor Professor Deng Yanchun found that the genome length is 9.33kb, located on chromosome 14q11.2, mRNA was 2024bp in length, encoding 357 amino acid residues with a molecular weight of about 40.79kD, so it was named as P40 and SYLD. homology analysis indicated that the NDRG1 (also known as brain development related protein) are highly homologous and contains an acyl carrier protein domain, it is known as the ndrg2. study results show that the high expression of NDRG2 in normal in the organization, but low expressed in some tumor tissues and tumor cell lines without expression, suggesting that the gene may be related to the occurrence and development of tumor, therefore NDRG2 may be a new candidate tumor suppressor gene. A recent study showed that NDR G2 is involved in the development of depression and Alzheimer's disease and other brain diseases, the expression of NDRG2 is regulated by adrenal steroids and anti depressant drugs, the study also found that NDRG2 is widely expressed in astrocytes. Our recent research found that NDRG2 protein has a certain time and space in the process of mouse and human embryonic development the expression characteristics with tissue, organs mature expression is more significant and widespread. In addition NDRG2 protein in mouse postnatal development process, brain, liver, kidney and skeletal muscle has a certain time and space expression characteristics of various tissues, NDRG2 protein is widely expressed in adult mice. Hou Shuangxing research showed that the expression of NDRG2 has certain characteristics time and space in the process of fetal brain development, NDRG2 is widely expressed in the brain of fetal brain function area, and with the development of brain maturation, expression is more significant, so as to push NDRG2 may be on the development of nerve cells, mature has important influence, like NDRG1 is a kind of brain development related gene, but its expression in postnatal brain development in different stages has not been investigated; and the other as the expression system development related gene NDRG2 in the central nervous system and the development process of what the law is. Not clear.
[Objective]:
1. to investigate the expression of NDRG2 in the forebrain of rats at different ages after birth.
2. to investigate the expression of NDRG2 in the respiratory system of human embryo at different gestational ages for 16~28 weeks.
[research methods]:
1. immunofluorescence double labeling method was used to determine the expression and location of NDRG2 at the protein level in the forebrain of rats of different ages.
From 1 days (neonatal period), 60 days (youth stage) and 240 days (middle age), the forebrain tissues of rats were observed by NDRG2 and GFAP immunofluorescence double labeling method. The expression pattern (number, morphology and distribution) of NDRG2 protein in the forebrain of rats and the location relationship between NDRG2 and GFAP were observed.
2. immunohistochemical method was used to determine the expression of NDRG2 in the 16~28 week human embryonic respiratory system and the location of the cells from the protein level.
Collect 16~28 weeks abortion fetal respiratory lung and trachea tissue, made of paraffin, by anti NDRG2 monoclonal antibody and immunohistochemical staining (ABC method), from the observation of NDRG2 protein expression. The statistical number of positive cells, using statistical methods to determine the different gestational age of lung and trachea between the expression of NDRG2 has no the difference.
[results]:
1. NDRG2 visible in the globus pallidus, cortex, piriform cortex, dorsal thalamus, hypothalamus, choroid plexus and other parts are widely expressed in the expression of immunoreactive products in the gray matter than white matter. It is worth noting that in 1 days and 60 days in rat forebrain, NDRG2 protein was mainly expressed in the nerve the cytoplasm of cells, ependymal cells and epithelial cells of the choroid plexus in the room, not observed with co expression of GFAP; while in the brain tissue of rats in 240 days, in the globus pallidus, cortical, optic nerve, dorsal thalamus and hypothalamus co expression of NDRG2 protein and GFAP positive cells were visible. In addition this study for the first time 3 ages of rat supraoptic nucleus can be observed in cells expressing NDRG2 protein.
2. NDRG2 was widely expressed in the 16~28 week embryonic respiratory system. The positive products were mainly located in cytoplasm of epithelial cells, but there was no significant difference in NDRG2 expression between different gestational ages.
[Conclusion]:
The 1. NDRG2 protein of nerve cells in 1 days and 60 days in rat forebrain may be widely expressed, NDRG2 as a development related gene, widely participate in the growth and development of the central nervous system. Co expression in 240 day rat forebrain observed NDRG2 protein and GFAP, and in 1 days and 60 days in the rat brain without observed NDRG2 protein and GFAP expression, suggesting that astrocytes in the forebrain of middle-aged rats the expression of the NDRG2. in addition, this is the first study found that expression of NDRG2 protein in nerve cells were 3 ages of rat supraoptic nucleus.
2. NDRG2铔嬬櫧鍦，

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