TISSUE CULTURE MODEL FOR RETINITIS PIGMENTOSA--A MOLECULAR APPROACH

每年有超过一半的美国人失明 都被诊断出有遗传缺陷 眼部疾病， 视网膜色素上皮（PE）与视网膜色素变性有关 (RP)、回旋状萎缩、脉络膜、脉络膜硬化和Stargedt's 黄斑变性该项目的长期目标是确定 并表征遗传缺陷和/或眼部疾病中的缺陷 广泛地归类为视网膜色素变性（RP）。 视网膜色素变性是一种 以最终丧失 感光细胞 正常和营养不良动物模型的研究 提示RP患者视力障碍可能继发于 视网膜和色素上皮（PE）。 虽然RP是最常见的 发生视网膜变性疾病，病因和 疾病的发病机制既缺乏又令人困惑。 可用信息的缺陷可能与以下因素有关： 生物组织的可用性不足。 原代外植体培养物 PE的视网膜具有有限的寿命， 去分化 因此，它们不是该研究的最佳模型 视网膜退化 该提案的目标是追求一个 另一种方法，并建立眼细胞系从正常和RP 供体眼（常染色体隐性遗传、常染色体显性遗传和伴性遗传）， 使用多种原癌基因和病毒DNA（H-ras -（瓦尔12）v- myc、SV-40病毒DNA和SV-40 T抗原基因。 我们已经 利用原癌基因建立了多种正常PE细胞系 (see附录），并已能够获得更多的差异化 通过使用细胞外基质自发建立的细胞系 (see附录）。 在本研究期间，我们建议建立视网膜 RP供体的细胞系，并比较正常永生细胞系 与RP细胞系的生长、形态和PE - 视杆细胞外节和视网膜的吞噬作用等功能标记物 新陈代谢. 我们还将探讨生长因子的作用， 感光细胞分化中的化学诱导剂，如RP， 最终导致感光细胞退化的疾病。 长期 我们的目标将是通过使用该工具识别和纠正RP中的病变， 组织培养方法。

More than half of the thousands of Americans who become blind each year are diagnosed as having a genetic defect. Ocular diseases in which retinal pigment epithelium (PE) is implicated are retinitis pigmentosa (RP), gyrate atrophy, choroidemia, choroidal sclerosis, and Stargedt's macular degeneration. The long-term goal of this project is to identify and characterize the genetic defect and/or defects in ocular diseases broadly grouped as retinitis pigmentosa (RP). Retinitis pigmentosa is a degenerative retinal disease characterized by eventual loss of photoreceptor cells. Studies on normal and dystrophic animal models suggest that visual failure in RP may be secondary to dysfunction in the retina and pigment epithelium (PE). Although RP is the commonest occurring retinal degenerative disease, the information on etiology and mechanism of pathogenesis in disease is both scant and confusing. Deficiencies in available information could be singularly related to inadequate availability of biological tissues. Primary explant cultures of PE, retina have a finite lifespan and a few permanent cell lines are dedifferentiated. As such they are not an optimal model for the study of retinal degeneration. The goal of this proposal is to pursue an alternate approach and establish ocular cell lines from normal and RP donor eyes (autosomal recessive, autosomal dominant and sex linked) by use of a variety of protooncogenes and viral DNAs (H-ras - (Val 12) v- myc, SV-40 viral DNA and SV-40T antigen gene. We have already established a variety of normal PE cell lines by use of protooncogenes (see Appendix) and have also been able to obtain more differentiated spontaneously established cell lines by use of extracellular matrices (see Appendix). In this study period, we propose to establish retinal cell lines from RP donors and compare the normal immortal cell lines with RP cell lines for growth, morphology and expression of PE - functional markers like phagocytosis of rod outer segments and retinal metabolism. We will also explore the role of growth factors and chemical inducers in photoreceptor cell differentiation as RP is ultimately a disease of photoreceptor cell degeneration. The long-term goal will be to identify and correct the lesion in RP by using this tissue culture approach.