【作者】 张姬慧；

【导师】 朱益华； 徐国兴；

【作者基本信息】 福建医科大学 ， 眼科学， 2008， 硕士

【摘要】 目的:研究蛋白激酶C三种同工酶PKCα,βⅡ,δ在正常大鼠出生后视网膜发育过程中的差异表达情况。方法:采用实时荧光定量逆转录-聚合酶链反应(real time RT-PCR)方法检测正常大鼠视网膜发育过程中PKCα,βⅡ,δmRNA水平的表达差异;通过免疫组织化学的方法检测PKCα,βⅡ,δ蛋白水平的表达情况。结果:Real time RT-PCR结果显示,大鼠出生后14d(P14)之前,视网膜组织中PKCα,βⅡmRNA表达逐渐升高, P14之后有所下降;PKCδ在生后早期高表达;成年后3种同工酶mRNA表达稳定。视网膜切片免疫组织化学染色光密度测定结果与mRNA表达相符。染色阳性产物定位显示,出生后当天(P0)PKCα,βⅡ阳性产物染色不明显,PKCδ阳性产物在原始神经母细胞层和原始节细胞层均有表达。随着视网膜的发育,生后7d(P7)PKCα染色阳性产物在外丛状层和内核层染色明显;PKCβⅡ的阳性产物主要在内核层、内丛状层、外丛状层和神经节细胞层染色明显;PKCδ的阳性产物主要分布在外界膜和视锥、视杆层。随着视网膜的发育,到了出生后14d(P14)PKCα可以在外丛状层看到轮廓清晰的强染色细胞,βⅡ染色阳性产物在神经节细胞层染色加深,内核层、内丛状层、外丛状层染色变浅;PKCδ在外界膜染色加深;视网膜发育成熟后,3种同工酶的阳性染色部位不变。结论:PKCα,βⅡ,δ在大鼠出生后视网膜发育过程中均有表达,并有一定的发育规律性,提示这3种同工酶与大鼠生后视网膜的发育过程密切相关;PKCα,βⅡ,δ表达的差异性提示:每种同工酶在参与视网膜发育方面可能具有不同的功能。PKCα可能对视杆-双极细胞发育有重要意义;PKCβⅡ可能对视网膜神经节细胞的发育过程有重要作用。PKCδ可能与视网膜生后早期原始神经母细胞和原始神经节细胞的分化及细胞凋亡过程有关。更多还原

【Abstract】 Objective:To investigate the exprepression and distribution of protein kinase Cα,βⅡ,δin the retinal during the development of poatnatal hamster.Methods: Real time RT-PCR method was used to detect the expression of mRNA;Immunocytochemical menthod was used to detect peotein.Results : The rusult of Real time RT-PCR showed that PKCα,βⅡmRNA was continual rise before 14th day of poatnatal development and then reduced till the lowest point in the adulthood. PKCδwas highly express in the early period of poatnatal development.The result of of spectrodensitometry was match to the result of Real time RT-PCR . In the first day(P0), the expression of PKCα,βⅡwas poor , the expression of PKCδwas found in neuroblastic layer and ganglion cell layer.With the development, at the seventh day, PKCαwas pronoounced in the external plexiform layer and inner nuclear layer; PKCβⅡwas pronoounced in the inner nuclear layer,inner plexiform layer, external plexiform layer and ganglion cell layer; PKCδwas pronoounced in the external limiting membrane and cone and rod layer.At the 14th day, isozymes were more localized than before. With mature the three isoenyme distribute invariably.Conclution: Protein kinase Cα,βⅡ,δwere expressed regularly in the whole process of rat retina development. It is suggest that the three isoenzyme is closely related to the early development of ratina in the postnatal hamster;The changed expression of PKCα,βⅡ,δis suggested as: each isoenzyme may have different roles in processing of rat ratinal development. PKCαmay be important to retinal rod-bipolar cells development; PKCβⅡmay be important to ratinal ganglion cells; PKCδmay participate the process of differentiation and Apoptosis of Primitive neuroblastic cells and Primitive ganglion cells in the early period .更多还原