本文選題：BMP-4 + 少突膠質(zhì)前體細(xì)胞��； 參考：《復(fù)旦大學(xué)》2010年碩士論文

【摘要】： 在視覺系統(tǒng)發(fā)育過程中,來源于腦的少突膠質(zhì)前體細(xì)胞匯集于視交叉處,并繼續(xù)沿著視神經(jīng)向視網(wǎng)膜方向遷移。這些細(xì)胞在胚胎后期至生后早期分化為少突膠質(zhì)細(xì)胞,并包裹節(jié)細(xì)胞軸突形成有髓神經(jīng)纖維。然而,大多數(shù)哺乳動(dòng)物的視神經(jīng)纖維進(jìn)入篩板后,其髓鞘即消失。究其原因,一般認(rèn)為可能與篩板處存在抑制少突膠質(zhì)前體細(xì)胞遷移的特殊屏障有關(guān),但具體機(jī)制仍有待深入研究。 目的通過研究BMP-4在視神經(jīng)及視網(wǎng)膜中的表達(dá)情況,并通過體外實(shí)驗(yàn)研究BMP-4對少突膠質(zhì)前體細(xì)胞分化的影響,及BMP-4與Olig2的相互關(guān)系,為視網(wǎng)膜及視神經(jīng)乳頭不形成髓鞘的機(jī)制提供實(shí)驗(yàn)依據(jù),為相關(guān)髓鞘疾病的治療提供新的思路。 方法用免疫組織化學(xué)染色檢測BMP-4在視網(wǎng)膜及視神經(jīng)乳頭中的表達(dá)及分布情況。采用恒溫振蕩法和差速貼壁法分離純化少突膠質(zhì)前體細(xì)胞,并研究BMP-4對少突膠質(zhì)前體細(xì)胞分化發(fā)育的影響。用Western Blot研究BMP-4與Olig2的相互關(guān)系。 結(jié)果在發(fā)育階段中,BMP-4僅選擇性表達(dá)在視神經(jīng)乳頭處。成功純化少突膠質(zhì)前體細(xì)胞,當(dāng)將培養(yǎng)液中的促細(xì)胞增殖因子撤除后,少突膠質(zhì)前體細(xì)胞自發(fā)向少突膠質(zhì)細(xì)胞分化和成熟,而加入BMP-4后,少突膠質(zhì)前體細(xì)胞的分化方向發(fā)生變化,高濃度的BMP-4使少突膠質(zhì)前體細(xì)胞最終分化為Ⅱ型星形膠質(zhì)細(xì)胞。隨著BMP-4濃度的增高,蛋白Olig2的表達(dá)量也逐漸降低。 結(jié)論在視神經(jīng)乳頭處高濃度表達(dá)的BMP-4蛋白有可能作用于遷移至篩板處的少突膠質(zhì)前體細(xì)胞。BMP-4在體外可抑制少突膠質(zhì)前體細(xì)胞向少突膠質(zhì)細(xì)胞的分化,并促使其向星形膠質(zhì)細(xì)胞分化。通過激活少突膠質(zhì)前體細(xì)胞內(nèi)的相關(guān)信號(hào)途徑,下調(diào)少突膠質(zhì)前體細(xì)胞內(nèi)Olig2蛋白的表達(dá)水平,從而抑制少突膠質(zhì)前體細(xì)胞向少突膠質(zhì)細(xì)胞方向分化,并促進(jìn)其向星形膠質(zhì)細(xì)胞方向分化。
[Abstract]:During the development of the visual system, oligodendrocyte precursor cells from the brain converge at the optic chiasma and continue to migrate along the optic nerve to the retina.These cells differentiate into oligodendrocytes from late embryo to early postnatal stage and form myelinated nerve fibers by wrapping axons of ganglion cells.However, when the optic nerve fibers of most mammals enter the sieve plate, their myelin sheath disappears.It is generally believed that there is a special barrier at the sieve plate to inhibit the migration of oligodendrocyte precursor cells, but the specific mechanism remains to be further studied.Objective to study the expression of BMP-4 in optic nerve and retina, and to investigate the effect of BMP-4 on differentiation of oligodendrocyte precursor cells in vitro, and the relationship between BMP-4 and Olig2.To provide experimental basis for the mechanism of no myelin sheath in retina and optic papilla and to provide new ideas for the treatment of related myelin diseases.Methods Immunohistochemical staining was used to detect the expression and distribution of BMP-4 in retina and optic papilla.Oligodendrocyte precursor cells were isolated and purified by isothermal oscillation and differential adherent method, and the effects of BMP-4 on differentiation and development of oligodendrocyte precursor cells were studied.The relationship between BMP-4 and Olig2 is studied by Western Blot.Results BMP-4 was only selectively expressed in the optic papilla during development.Oligodendrocyte progenitor cells were successfully purified. After removing the proliferating factor from the culture medium, oligodendrocyte progenitor cells spontaneously differentiated and matured into oligodendrocytes, but after adding BMP-4, the oligodendrocyte progenitor cells spontaneously differentiated and matured into oligodendrocytes.The differentiation direction of oligodendrocyte precursor cells changed, and high concentration of BMP-4 resulted in the differentiation of oligodendrocyte precursor cells into type 鈪，

本文編號(hào)：1744777