本文選題：ndrg2　切入點(diǎn)：大鼠前腦　出處：《第四軍醫(yī)大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

【摘要】： 【研究背景】 Ndrg2與ndrg1、ndrg3和ndrg4共同構(gòu)成了一個(gè)新的基因家族,即ndrg家族。該基因家族參與了細(xì)胞的生長(zhǎng),但具體機(jī)制仍不清楚。人ndrg2由我的導(dǎo)師鄧艷春教授發(fā)現(xiàn),其基因組全長(zhǎng)為9.33kb,位于染色體的14q11.2,mRNA全長(zhǎng)2024bp,編碼含357個(gè)氨基酸殘基,分子量約為40.79kD,因此曾命名為p40和SYLD。同源性分析表明,其與ndrg1(也稱做腦發(fā)育相關(guān)蛋白)具有高度同源性,并含有一個(gè)�；鶖y帶蛋白結(jié)構(gòu)域,故稱為ndrg2。研究結(jié)果初步顯示,ndrg2在多種正常組織中高表達(dá),而在某些腫瘤組織和腫瘤細(xì)胞系中低表達(dá)或無(wú)表達(dá),推測(cè)該基因可能與腫瘤的發(fā)生發(fā)展有關(guān),因此提出ndrg2可能為新的抑癌候選基因。新近資料表明,ndrg2參與了抑郁癥和阿爾茨海默氏病等腦部疾病的發(fā)生發(fā)展,NDRG2的表達(dá)還受腎上腺類固醇和抗抑郁藥物的調(diào)節(jié),進(jìn)行該研究時(shí)還發(fā)現(xiàn)NDRG2廣泛表達(dá)于星形膠質(zhì)細(xì)胞。我室近期研究發(fā)現(xiàn)NDRG2蛋白在小鼠和人胚胎發(fā)育過(guò)程中具有一定的時(shí)間和空間表達(dá)特性,隨著組織、器官的成熟表達(dá)更為顯著和廣泛。此外NDRG2蛋白在小鼠生后發(fā)育過(guò)程中心、腦、肝、腎和骨骼肌中具有一定時(shí)間和空間表達(dá)特性,NDRG2蛋白廣泛表達(dá)于成年小鼠的多種組織。侯雙興等研究表明NDRG2在胎腦發(fā)育過(guò)程中具有一定時(shí)間空間的表達(dá)特性,認(rèn)為NDRG2在胎腦各腦功能區(qū)中廣泛表達(dá),并隨著腦發(fā)育的成熟,表達(dá)更為顯著,從而推測(cè)ndrg2可能對(duì)神經(jīng)細(xì)胞發(fā)育、成熟有重要影響,同ndrg1一樣是一種腦發(fā)育相關(guān)基因,但其在出生后腦發(fā)育各階段的表達(dá)情況還沒(méi)有人研究;而且作為發(fā)育相關(guān)基因ndrg2在中樞神經(jīng)系統(tǒng)外的其他系統(tǒng)發(fā)育過(guò)程的表達(dá)有什么規(guī)律,目前也不清楚。 【研究目的】: 1.探討NDRG2在出生后不同年齡大鼠前腦的表達(dá)規(guī)律。 2.探討NDRG2在不同胎齡16～28周人胚胎呼吸系統(tǒng)的表達(dá)情況。 【研究方法】: 1.免疫熒光雙重標(biāo)記法從蛋白水平確定NDRG2在不同年齡大鼠前腦的表達(dá)特點(diǎn)及細(xì)胞定位。 取1天(新生兒期)、60天(青年期)及240天(中年期)大鼠前腦組織,用NDRG2與GFAP免疫熒光雙重標(biāo)記法觀察NDRG2蛋白在大鼠前腦的表達(dá)規(guī)律(數(shù)目、形態(tài)及分布)及NDRG2同GFAP的位置關(guān)系。 2.免疫組織化學(xué)法從蛋白水平確定NDRG2在16～28周人胚胎呼吸系統(tǒng)的表達(dá)特點(diǎn)及細(xì)胞定位。 搜集16～28周人工流產(chǎn)胎兒呼吸系統(tǒng)的肺和氣管組織,制成石蠟切片,用抗NDRG2單克隆抗體,行免疫組化染色(ABC法),從蛋白水平觀察NDRG2表達(dá)情況。統(tǒng)計(jì)陽(yáng)性細(xì)胞數(shù),利用統(tǒng)計(jì)學(xué)方法,判斷不同胎齡的肺和氣管間NDRG2表達(dá)有無(wú)差別。 【結(jié)果】: 1.可見(jiàn)NDRG2在蒼白球、皮質(zhì)、梨狀皮質(zhì)、背側(cè)丘腦、下丘腦、脈絡(luò)叢等部位有廣泛的表達(dá),免疫陽(yáng)性產(chǎn)物在灰質(zhì)中的表達(dá)多于白質(zhì)。值得注意的是在1天及60天大鼠的前腦中,NDRG2蛋白主要表達(dá)于神經(jīng)細(xì)胞、室管膜細(xì)胞及脈絡(luò)叢上皮細(xì)胞的胞漿中,未觀察到與GFAP共表達(dá);而在240天的大鼠腦組織中,在蒼白球、皮質(zhì)、視神經(jīng)、背側(cè)丘腦及下丘腦可見(jiàn)NDRG2蛋白與GFAP陽(yáng)性細(xì)胞共表達(dá)。此外本研究首次發(fā)現(xiàn)3個(gè)年齡段的大鼠視上核神經(jīng)細(xì)胞內(nèi)均可觀察到NDRG2蛋白的表達(dá)。 2. NDRG2在16～28周胚胎呼吸系統(tǒng)中有廣泛的表達(dá),陽(yáng)性產(chǎn)物主要位于上皮細(xì)胞的胞漿中,但是不同胎齡之間NDRG2的表達(dá)未見(jiàn)明顯差別。 【結(jié)論】: 1. NDRG2蛋白在1天及60天大鼠前腦內(nèi)的神經(jīng)細(xì)胞廣泛表達(dá),可能是ndrg2作為發(fā)育相關(guān)基因,廣泛參與中樞神經(jīng)系統(tǒng)生長(zhǎng)發(fā)育的結(jié)果。在240天大鼠前腦觀察到NDRG2蛋白與GFAP共表達(dá),而在1天及60天大鼠腦內(nèi)未觀察到NDRG2蛋白與GFAP共表達(dá),提示中年大鼠前腦內(nèi)星形膠質(zhì)細(xì)胞也表達(dá)NDRG2。此外,本研究首次發(fā)現(xiàn)NDRG2蛋白在3個(gè)年齡段大鼠下丘腦視上核的神經(jīng)細(xì)胞均有表達(dá)。 2. NDRG2蛋白在16～28周人胚胎呼吸系統(tǒng)的假?gòu)?fù)層柱狀纖毛上皮和單層立方上皮有廣泛的表達(dá),提示ndrg2在中、末期胚胎的呼吸系統(tǒng)上皮細(xì)胞的生長(zhǎng)與發(fā)育過(guò)程中起一定作用。NDRG2陽(yáng)性產(chǎn)物定位于上皮細(xì)胞的胞漿,提示NDRG2可能是一種胞漿蛋白。
[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，

本文編號(hào)：1561129