Abstract
PurposeTo investigate the role of Pou4f1 and Pou4f2 in the survival of adult retinal ganglion cells (RGCs).MethodsConditional alleles of Pou4f1 and Pou4f2 were generated (Pou4f1loxP and Pou4f2loxP respectively) for the removal of Pou4f1 and Pou4f2 in adult retinas. A tamoxifen-inducible Cre was used to delete Pou4f1 and Pou4f2 in adult mice and retinal sections and flat mounts were subjected to immunohistochemistry to confirm the deletion of both alleles and to quantify the changes in the number of RGCs and other retinal neurons. To determine the effect of loss of Pou4f1 and Pou4f2 on RGC survival after axonal injury, controlled optic nerve crush (CONC) was performed and RGC death was assessed.Results Pou4f1 and Pou4f2 were ablated two weeks after tamoxifen treatment. Retinal interneurons and Müller glial cells are not affected by the ablation of Pou4f1 or Pou4f2 or both. Although the deletion of both Pou4f1 and Pou4f2 slightly delays the death of RGCs at 3 days post-CONC in adult mice, it does not affect the cell death progress afterwards. Moreoever, deletion of Pou4f1 or Pou4f2 or both has no impact on the long-term viability of RGCs at up to 6 months post-tamoxifen treatment.Conclusion Pou4f1 and Pou4f2 are involved in the acute response to damage to RGCs but are dispensable for the long-term survival of adult RGC in mice.
Highlights
Glaucoma, a retinal degeneration disease characterized by progressive loss of retinal ganglion cells (RGCs), affected over 60 million people worldwide in 2010 and the number will increase to about 80 million by 2020 [1]
In order to investigate the role of POU4F factors in adult RGCs, we focused on POU4F1 and POU4F2 whose combined expression covers almost all RGCs in the adult retina
In order to investigate the role of Pou4f1 and Pou4f2 in adult RGCs, we deleted Pou4f1 and Pou4f2 by intraperitoneal injection of tamoxifen at P30 at the dosage of 5 mg/40 g bodyweight for five consecutive days
Summary
A retinal degeneration disease characterized by progressive loss of retinal ganglion cells (RGCs), affected over 60 million people worldwide in 2010 and the number will increase to about 80 million by 2020 [1]. As the second leading cause of blindness, glaucoma is responsible for millions of blindness worldwide and most clinical glaucoma cases are in advanced conditions due to the inconspicuous early symptoms and the lack of effective early diagnosis. Pou4f2 expression starts in more than 80% RGC precursors at embryonic day 11.5 (E11.5), a time when RGCs are first generated [2,4]. Afterwards, Pou4f1 and Pou4f3 are expressed in 80% and 20% developing RGCs, respectively [2,5,6]
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