Discipline of Ophthalmology, Sydney Medical School, The University of Sydney, Save Sight Institute, Sydney, New South Wales, Australia.
Department of Ophthalmology and Biochemistry, West Virginia University, Morgantown, West Virginia, USA.
Glia. 2021 Aug;69(8):1966-1986. doi: 10.1002/glia.24005. Epub 2021 Apr 9.
The importance of Müller glia for retinal homeostasis suggests that they may have vulnerabilities that lead to retinal disease. Here, we studied the effect of selectively knocking down key metabolic genes in Müller glia on photoreceptor health. Immunostaining indicated that murine Müller glia expressed insulin receptor (IR), hexokinase 2 (HK2) and phosphoglycerate dehydrogenase (PHGDH) but very little pyruvate dehydrogenase E1 alpha 1 (PDH-E1α) and lactate dehydrogenase A (LDH-A). We crossed Müller glial cell-CreER (MC-CreER) mice with transgenic mice carrying a floxed IR, HK2, PDH-E1α, LDH-A, or PHGDH gene to study the effect of selectively knocking down key metabolic genes in Müller glia cells on retinal health. Selectively knocking down IR, HK2, or PHGDH led to photoreceptor degeneration and reduced electroretinographic responses. Supplementing exogenous l-serine prevented photoreceptor degeneration and improved retinal function in MC-PHGDH knockdown mice. We unexpectedly found that the levels of retinal serine and glycine were not reduced but, on the contrary, highly increased in MC-PHGDH knockdown mice. Moreover, dietary serine supplementation, while rescuing the retinal phenotypes caused by genetic deletion of PHGDH in Müller glial cells, restored retinal serine and glycine homeostasis probably through regulation of serine transport. No retinal abnormalities were observed in MC-CreER mice crossed with PDH-E1α- or LDH-A-floxed mice despite Cre expression. Our findings suggest that Müller glia do not complete glycolysis but use glucose to produce serine to support photoreceptors. Supplementation with exogenous serine is effective in preventing photoreceptor degeneration caused by PHGDH deficiency in Müller glia.
Müller 胶质细胞对视网膜内稳态的重要性表明,它们可能存在易损性,从而导致视网膜疾病。在这里,我们研究了选择性敲低 Müller 胶质细胞中关键代谢基因对光感受器健康的影响。免疫染色表明,鼠 Müller 胶质细胞表达胰岛素受体 (IR)、己糖激酶 2 (HK2) 和磷酸甘油酸脱氢酶 (PHGDH),但丙酮酸脱氢酶 E1α1 (PDH-E1α) 和乳酸脱氢酶 A (LDH-A) 表达很少。我们将 Müller 胶质细胞-CreER (MC-CreER) 小鼠与携带 IR、HK2、PDH-E1α、LDH-A 或 PHGDH 基因的 floxed 转基因小鼠杂交,以研究选择性敲低 Müller 胶质细胞中关键代谢基因对视网膜健康的影响。选择性敲低 IR、HK2 或 PHGDH 导致光感受器变性和视网膜电图反应降低。补充外源性 l-丝氨酸可预防 MC-PHGDH 敲低小鼠的光感受器变性并改善视网膜功能。我们出人意料地发现,MC-PHGDH 敲低小鼠的视网膜丝氨酸和甘氨酸水平没有降低,反而显著升高。此外,饮食丝氨酸补充虽然挽救了 Müller 胶质细胞中 PHGDH 基因缺失引起的视网膜表型,但通过调节丝氨酸转运,恢复了视网膜丝氨酸和甘氨酸的内稳态。尽管 Cre 表达,但在与 PDH-E1α 或 LDH-A-floxed 小鼠杂交的 MC-CreER 小鼠中未观察到视网膜异常。我们的研究结果表明,Müller 胶质细胞不完全进行糖酵解,而是利用葡萄糖产生丝氨酸来支持光感受器。补充外源性丝氨酸可有效预防 Müller 胶质细胞中 PHGDH 缺乏引起的光感受器变性。