Muller Cell Domains in the Retina

视网膜中的米勒细胞域

• 批准号: 6958449
• 负责人: DAVID A DILORETO
• 金额: $ 18.01万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6958449
• 关键词: Adenoviridae; Muller' s cell; acid base balance; astrocytes; biological signal transduction; calcium flux; cell cell interaction; cell morphology; cell population study; confocal scanning microscopy; developmental neurobiology; gliosis; green fluorescent proteins; laboratory rat; neurons; neuropathology; organ culture; photostimulus; retina circulation; retina degeneration; tissue /cell culture; transfection /expression vector; visual photoreceptor

DESCRIPTION (provided by applicant): Defining Muller cell domains and the effect that their breakdown has on the retina is an entirely new approach to studying retinal degenerations. Defining an anatomic Muller cell domain has implications for each cell that it contacts, while the functionality of the domain has implications for the Muller cell's role as an active regulator of neuronal processing. If the Muller cell's domain is compromised during degeneration, this could result in aberrant glial and neuronal signaling, insufficient delivery of substrates and/or less efficient removal of waste products, which would ultimately contribute to neuronal degeneration. My goal in this proposal is to test the hypotheses that: (1) the Muller cell forms distinct anatomical domains; (2) domains have functional significance; and (3) breakdown of domains during degeneration contributes to neuronal dysfunction. To label individual Muller cells in vivo, intravitreal injections of adenovirus; will be used to deliver enhanced green fluorescent protein to the Muller cell. Confocal microscopy will be used to image the Muller cell and identify its domain within the retina. Once the domain is anatomically defined, calcium imaging studies will be performed via the caging and uncaging of calcium within Muller cells. Calcium signaling in adjacent Muller cells as well as photoreceptor cells will be measured. Changes in light-evoked photoreceptor cell responses within the Muller cell domain as well as pH will be measured and compared to changes outside the domain. Finally, a model of retinal degeneration will be used to test the above effects in Muller cells that have become reactive or gliotic. The role the Muller cell plays in health and in disease will be determined based on domain structure within the retina. How this affects photoreceptor cell function will also studied. Thus, establishing the central role that the Muller cell plays in retinal health and disease may provide a potential target for new clinical therapies that might delay or prevent neuronal loss and help to preserve vision.

描述（由申请人提供）：定义Muller细胞结构域以及其崩溃对视网膜的影响是研究视网膜退化的一种全新方法。定义解剖学的穆勒细胞结构域对其接触的每个细胞都具有影响，而域的功能对穆勒细胞作为神经元处理的活性调节剂的作用具有影响。如果在变性过程中穆勒细胞的结构域受到损害，则可能导致神经胶质和神经元信号传导异常，底物的递送不足和效率较低的废物去除，这最终会导致神经元退化。我在此提案中的目标是检验：（1）穆勒细胞形成不同的解剖域； （2）域具有功能意义； （3）变性过程中的域故障导致神经元功能障碍。为了在体内标记单个muller细胞，玻璃体内注射腺病毒；将用于将增强的绿色荧光蛋白传递到Muller细胞。共聚焦显微镜将用于成像Muller细胞并在视网膜内识别其域。一旦结构域在解剖学上定义，钙成像研究将通过Muller细胞内钙的笼子和不老龄来进行。将测量相邻muller细胞以及感光细胞中的钙信号传导。将测量Muller细胞结构域内的光感受器细胞反应的变化以及pH的变化，并将其与域外部的变化进行比较。最后，将使用视网膜变性模型来测试已反应性或胶质化的muller细胞中的上述效果。穆勒细胞在健康和疾病中所起的作用将根据视网膜内的域结构确定。这也将研究如何影响感光细胞功能。因此，确定穆勒细胞在视网膜健康和疾病中发挥作用的核心作用可能为新的临床疗法提供了潜在的靶标，这些临床疗法可能会延迟或预防神经元丧失并帮助保持视力。