Liu Li, Steinle Jena J
Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, MI, United States of America.
Department of Ophthalmology, Wayne State University School of Medicine, Detroit, MI, United States of America.
PLoS One. 2017 Dec 29;12(12):e0190253. doi: 10.1371/journal.pone.0190253. eCollection 2017.
Müller cells are key to metabolic and ionic regulation in the retina. They also produce a number of inflammatory mediators and are significantly affected in diabetic retinopathy. To investigate the role of toll-like receptor 4 (TLR4) in retinal Müller cells, we crossed TLR4 floxed with PDGFRα-Cre mice to eliminate TLR4 in retinal Müller cells. We performed Western blotting and ELISA analyses to determine whether loss of TLR4 affected myeloid differentiation primary response protein (MyD88)-dependent or -independent signaling, leading to reduced levels of tumor necrosis factor alpha (TNFα) and interleukin 1 beta (IL1β) in whole retinal lysates from the TLR4 floxed and TLR4-PDGFRα-Cre mice. Data show that TLR4-PDGFRα-Cre mice have reduced levels of both the MyD88-dependent and -independent signaling pathways. These studies confirm successful development of a Müller cell-specific TLR4 knockout mouse colony. These mice have reduced MyD88-dependent and -independent signaling pathway proteins, as well as reduced TNFα and IL1β levels. These mice can be used to dissect TLR4 signaling in disorders affecting retinal Müller cells.
穆勒细胞对视网膜的代谢和离子调节至关重要。它们还会产生多种炎症介质，并且在糖尿病视网膜病变中受到显著影响。为了研究Toll样受体4（TLR4）在视网膜穆勒细胞中的作用，我们将携带floxed TLR4的小鼠与PDGFRα-Cre小鼠杂交，以消除视网膜穆勒细胞中的TLR4。我们进行了蛋白质免疫印迹和酶联免疫吸附测定分析，以确定TLR4的缺失是否影响髓样分化初级反应蛋白（MyD88）依赖性或非依赖性信号传导，从而导致来自携带floxed TLR4的小鼠和TLR4-PDGFRα-Cre小鼠的全视网膜裂解物中肿瘤坏死因子α（TNFα）和白细胞介素1β（IL1β）水平降低。数据显示，TLR4-PDGFRα-Cre小鼠的MyD88依赖性和非依赖性信号传导途径水平均降低。这些研究证实成功培育出了穆勒细胞特异性TLR4基因敲除小鼠群体。这些小鼠的MyD88依赖性和非依赖性信号传导途径蛋白水平降低，TNFα和IL1β水平也降低。这些小鼠可用于剖析影响视网膜穆勒细胞的疾病中的TLR4信号传导。
