本文選題：神經(jīng)管 + 發(fā)育�。� 參考：《山東大學》2007年碩士論文

【摘要】： 目的：組織和器官的形態(tài)發(fā)生(morphogenesis)是通過細胞的形態(tài)變化和運動、細胞的識別和粘附、細胞的增殖和凋亡等過程而實現(xiàn)的。神經(jīng)管的形成和發(fā)育也是一個極其復雜的生物學過程，包括細胞增殖、凋亡、分化、識別、遷移和功能的表達等等。近年來的研究表明：神經(jīng)管的發(fā)育不僅受到細胞基因表達的控制，而且還與胚胎和神經(jīng)上皮產(chǎn)生的多種調(diào)節(jié)因子有關，凝集素受體便是這些調(diào)節(jié)因子中的一員。 凝集素受體是一類分布于細胞膜表面的糖蛋白、糖脂或糖復合物。凝集素受體上具有能被凝集素特異識別并結合的糖基。我們以昆明小鼠為研究對象，用親和細胞化學的方法對正常鼠胚神經(jīng)管和過量維生素A酸致畸鼠胚神經(jīng)管發(fā)育中WGA、RCA-Ⅰ、ECL三種凝集素受體的分布狀況以及變化規(guī)律進行了研究，以期闡明上述三種凝集素受體在正常神經(jīng)管及過量維生素A酸致畸神經(jīng)管發(fā)育中的作用。 方法：將健康育齡雌性昆明小鼠與正常具有生育能力的雄鼠按2∶1合籠，次日晨查見陰栓者，當天中午定為孕0.5天。孕鼠受精后第7天下午6∶30一次性灌服花生油稀釋的維生素A酸30mg／kg，以獲取神經(jīng)管畸形的小鼠胚胎。將正常組孕鼠和維生素A酸致畸組的孕鼠分別于孕第7.5、8.5、9.0、9.5、10.5、11.5，12.5天頸椎脫臼處死并立即剖腹取出子宮。用刀片切取有膨大結節(jié)部位的子宮(含胚胎)；孕11.5d、12.5d的孕鼠在濾紙上沿子宮側軸將子宮剪開，仔細剝離柔軟的蛻膜、分離鼠胚。所得標本用Bouin固定液固定，常規(guī)石蠟包埋，做厚度為5μm的連續(xù)切片。采用SABC親和細胞化學法檢測WGA、RCA-Ⅰ、ECL三種凝集素受體在小鼠胚胎神經(jīng)管的分布情況。 結果： 1．WGA染色結果：在孕7.5～12.5天，WGA受體在神經(jīng)管的腔面、基底膜、神經(jīng)上皮細胞的胞膜和胞漿均有表達。WGA受體表達的總體趨勢是先下降后上升，其最低點出現(xiàn)于孕9.0天。在孕9.5～12.5天，WGA受體在頭部和尾部的表達水平明顯高于體部。在維生素A酸致畸組，WGA受體主要表達于神經(jīng)管的腔面，其它部位的表達減弱或消失。但是，在孕10.5～12.5天，WGA受體在神經(jīng)管的背部呈強陽性著色。無論神經(jīng)管的頭部、體部，還是尾部，WGA受體在致畸組表達水平明顯低于正常組。 2．RCA-Ⅰ染色結果：在孕7.5～12.5天，RCA-Ⅰ受體在神經(jīng)管的腔面、基底膜、神經(jīng)上皮細胞的胞膜和胞漿均有表達。在神經(jīng)管未關閉之前(孕7.5～9.0天)，RCA-Ⅰ受體的表達呈現(xiàn)逐漸上升的趨勢。在神經(jīng)管關閉之后(孕9.5～12.5天)，RCA-Ⅰ受體的表達在神經(jīng)管頭部和尾部仍呈現(xiàn)逐漸上升的趨勢，在神經(jīng)管體部，卻呈現(xiàn)逐漸下降的趨勢，而且，頭部和尾部的表達強度均高于體部。在維生素A酸致畸組，RCA-Ⅰ受體主要表達于神經(jīng)管的腔面，其它部位的表達減弱或消失。但在孕11.5～12.5天，神經(jīng)管的背部卻呈強陽性表達。無論神經(jīng)管的頭部、體部，還是尾部，RCA-Ⅰ受體在致畸組表達水平明顯低于正常組。 3．ECL染色結果：在孕7.5～12.5天，強陽性著色部位僅見于神經(jīng)管的腔面，基底膜、神經(jīng)上皮細胞的胞膜呈陽性、弱陽性或陰性著色，神經(jīng)上皮細胞的胞漿呈弱陽性或陰性著色。無論在神經(jīng)管的頭部、體部，，還是尾部，ECL受體的表達均呈現(xiàn)先下降后上升的趨勢，其最低點在孕9.0天。在維生素A酸致畸組，陽性著色部位與正常發(fā)育神經(jīng)管基本相同。但在孕11.5～12.5天，神經(jīng)管的背部卻呈強陽性表達。在孕9.5～12.5天神經(jīng)管的頭部，孕10.5～12.5天神經(jīng)管的尾部，孕11.5～12.5天神經(jīng)管的體部，ECL受體的表達水平低于正常發(fā)育的神經(jīng)管相應部位。 結論： 1．在神經(jīng)管正常發(fā)育過程中，三種凝集素受體的數(shù)量和分布狀況均存在時間和空間上的差異。 2．在神經(jīng)管關閉前和關閉后，三種凝集素受體的表達趨勢存在明顯差異。這可能與神經(jīng)管關閉的形態(tài)學事件有關。 3．在神經(jīng)管正常發(fā)育過程中，三種凝集素受體的表達均呈現(xiàn)規(guī)律性的變化。它們可能參與對神經(jīng)管發(fā)育的調(diào)節(jié)。 4．三種凝集素受體在神經(jīng)管表達水平的下降，尤其是WGA、RCA-Ⅰ表達水平的下降與維生素A酸致神經(jīng)管畸形相關。
[Abstract]:Objective: the morphogenesis of tissues and organs (morphogenesis) is realized through the process of cell morphological changes and movement, cell recognition and adhesion, cell proliferation and apoptosis. The formation and development of neural tubes are also an extremely complex biological process, including cell proliferation, apoptosis, differentiation, identification, migration, and function expression. In recent years, studies have shown that the development of the neural tube is not only controlled by the gene expression of the cells, but also related to a variety of regulatory factors produced by the embryo and the neuroepithelium. The agglutinin receptor is one of these regulatory factors.
Lectin receptor is a kind of glycoprotein, glycolipid or glycoconjugates on the surface of the cell membrane. The lectin receptor has a glycoprotein that can be specifically identified and combined by lectin. In Kunming mice, we used an affinity cytochemistry method for the development of WGA in the neural tube development of normal rat embryo neural tube and excessive vitamin A acid teratogenic mouse embryo. The distribution and variation of three kinds of lectin receptors of RCA- I and ECL were studied in order to elucidate the role of the three lectin receptors in the development of normal neural tube and excessive vitamin A acid teratogenic neural tube development.
Methods: the female Kunming mice of healthy childbearing age and the male mice with normal fertility were caged at 2: 1. The next day was found to be the 0.5 days of pregnancy. The pregnant rats were given the vitamin A acid 30mg / kg diluted by peanut oil at 6: 30 at seventh day after fertilization, in order to obtain the mouse embryos of the deformity of the God meridian. The pregnant rats in the vitamin A teratogenic group were sacrificed on the 7.5,8.5,9.0,9.5,10.5,11.5,12.5 day of the pregnancy and removed the uterus immediately. The uterus (containing the embryo) with the enlarged nodules was cut with the blade. The pregnant mice pregnant 11.5d and 12.5d cut the uterus along the lateral axis of the uterus on the filter paper and carefully stripped the soft decidua to separate the mouse embryos. The specimens were fixed with Bouin fixed solution, paraffin embedded and continuous slices with a thickness of 5 mu m. The distribution of WGA, RCA- I, ECL three agglutinin receptors in the neural tube of mouse embryos was detected by SABC affinity cytochemistry.
Result:
1.WGA staining results: at 7.5 to 12.5 days of pregnancy, the overall tendency of WGA receptor to express.WGA receptor expression in the cavity surface of the nerve canal, the basement membrane, the cytoplasm and the cytoplasm of the neuroepithelial cells was first descended and then increased, and the lowest point appeared at 9 days of pregnancy. The expression level of WGA receptor in the head and tail was significantly higher than that in the body at 9.5 to 12.5 days of pregnancy. In the vitamin A acid teratogenic group, the WGA receptor was mainly expressed in the cavity surface of the nerve canal, and the expression of other parts was weakened or disappeared. However, the WGA receptor was strongly stained on the back of the nerve tube at 10.5 to 12.5 days of pregnancy. No matter the head, body, or tail of the nerve canal, the expression level of WGA receptor in the teratogenic group was significantly lower than that of the normal group.
2.RCA- I staining results: at 7.5 to 12.5 days of pregnancy, the expression of RCA- I receptor in the cavity surface of the nerve canal, the basement membrane, the cell membrane and the cytoplasm of the neuroepithelial cells were expressed. The expression of RCA- I receptor showed a gradual rising trend before the nerve tube was closed (7.5 to 9 days of pregnancy). After the closure of the nerve canal (9.5 to 12.5 days), the table of RCA- I receptor At the head and tail of the nerve tube, the trend of the head and tail was gradually rising, but in the nerve tube body, the trend was gradually decreasing, and the expression intensity of the head and tail was higher than that in the body. In the vitamin A teratogenic group, the RCA- I receptor was mainly expressed in the cavity surface of the nerve canal, the expression of other parts was weakened or disappeared. But in the 11.5 to 12.5 days of pregnancy, The expression of RCA- I receptor in the teratogenic group was significantly lower than that in the normal group.
The results of 3.ECL staining: in the 7.5 to 12.5 day of pregnancy, the strong positive staining site was found only in the cavity surface of the nerve canal, the membrane of the basal membrane and the neuroepithelial cells was positive, weak positive or negative coloring, and the cytoplasm of the neuroepithelial cells was weak positive or negative coloring. The expression of the ECL receptor in the head, body and tail of the nerve tube appeared first. In the vitamin A acid teratogenic group, the positive staining site was basically the same as that of the normal developing nerve canal in the vitamin A teratogenic group. But the back of the nerve tube was strongly positive in the 11.5 to 12.5 day of pregnancy. The head of the nerve tube, the tail of the 10.5 to 12.5 day of the nerve canal, and the body of the nerve tube 11.5 to 12.5 days pregnant, and the body of the nerve tube in the 9.5 ~ 12.5 days of pregnancy. The expression level of ECL receptor was lower than that of normal neural tube.
Conclusion:
1. during the normal development of the neural tube, there are temporal and spatial differences in the number and distribution of the three lectin receptors.
2. there was a significant difference in the expression of three lectin receptors before and after the closure of neural tube. This may be related to the morphological events of neural tube closure.
3. during the normal development of neural tube, the expression of three lectin receptors showed regular changes. They may participate in the regulation of neural tube development.
4. the decrease of the expression of three lectin receptors in the neural tube, especially the decrease of WGA and RCA- I expression level, is related to the neural tube defects induced by vitamin A acid.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R363

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