title: Trans-Corneal Subretinal Injection in Mice and Its Effect on the Function and Morphology of the Retina.
abstract: OBJECTIVE
  To introduce a practical method of subretinal injection in mice and evaluate injection-induced retinal detachment (RD) and damage using a dynamic imaging system, electrophysiology, and histology.
  
  
  METHODS
  After full dilation of a 2-month-old C57BL/6J mouse pupil, the cornea near the limbus was punctured with a 30 ½-gague disposable beveled needle. A 33 ½-gauge blunt needle was inserted through the corneal perforation into the anterior chamber, avoiding the lens before going deeper into the vitreous cavity, and penetrating the inner retina to reach the subretinal space. The mice were divided into four groups: in group 1, about 80-100% of the retina was filled with subretinally injected solution; in group 2, approximately 50-70% of the retina was filled with injected solution; in group 3, the procedures were stopped before solution injection; and non-injected eyes were used as the negative control in group 4. An optical coherence tomography (OCT) imaging system was used to monitor retinal reattachment during the first three days following the injections. Histological and functional changes were examined by light microscopy and electroretinography (ERG) at five weeks post-injection.
  
  
  RESULTS
  After a short-term training, a 70% success rate with 50% or more coverage (i.e., retinal blebs occupied 50% or more retinal area and filled with the injected solution) with minimal injection-related damages can be achieved. Bleb formation was associated with retinal detachment (RD) between the neuroretina and the retinal pigment epithelium (RPE) layer. Partial RD could be observed at post-injection day 1, and by day 2 most of the retina had reattached. At 5 weeks post-injection, compared to uninjected control group 4, the b-wave amplitudes of ERG decreased 22% in group 1, 16% in group 2, and 7% in group 3; the b-wave amplitudes were statistically different between the uninjected group and the groups with either 50-70% or 80-100% coverage. The subretinal injection-induced RD reattached and became stable at five weeks post-injection, although some photoreceptor damage could still be observed in and around the injection sites, especially in 80-100% coverage group.
  
  
  CONCLUSIONS
  Trans-corneal subretinal injection is effective and practical, although subretinal injection-related damages can cause some morphological and functional loss.
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Yan
authors: Y
authors: Qi
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Xufeng
authors: X
authors: Dai
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Hua
authors: H
authors: Zhang
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Ying
authors: Y
authors: He
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Yangyang
authors: Y
authors: Zhang
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China; Fenyang College of Shanxi Medical University, Fenyang, Shanxi, P. R. China.
authors: Juanjuan
authors: J
authors: Han
authors: Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, Florida, United States of America.
authors: Ping
authors: P
authors: Zhu
authors: Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, Florida, United States of America; Department of Ophthalmology, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, P. R. China.
authors: Yuxin
authors: Y
authors: Zhang
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors: Qinxiang
authors: Q
authors: Zheng
authors: Department of Ophthalmology, First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, P. R. China.
authors: Xia
authors: X
authors: Li
authors: Department of Ophthalmology, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, P. R. China.
authors: Chen
authors: C
authors: Zhao
authors: School of Ophthalmology & Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China; Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, Florida, United States of America; Department of Ophthalmology, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, P. R. China.
authors: Jijing
authors: J
authors: Pang
mesh_terms: D000818:Animals; D003315:Cornea / Q000473:pathology / Q000502:physiology; D056965:Injections, Intraocular / Q000009:adverse effects* / Q000295:instrumentation / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D009339:Needles / Q000009:adverse effects; D012160:Retina / Q000473:pathology / Q000502:physiology; D012163:Retinal Detachment / Q000209:etiology*