Effect of congenital stationary night blindness on oxygen induced retinal neovascularization by improved Chinese ink perfusion method

Authors: Song Meixia,  Li Shang,  Niu Chao,  Zhang Zuoming
DOI: 10.3760/cma.j.issn.2095-0160.2017.10.006
Published 2017-10-10
Cite as Chin J Exp Ophthalmol, 2017,35(10): 885-889.

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Congenital stationary night blindness (CSNB) is a common genetic eye disease.Retinal angiogenesis is rarely obtained in retinal degeneration animal.The effects of CSNB on retinal angiogenesis require further study.


This study was to improve Chinese ink perfusion technology and to explore the effect of CSNB on oxygen induced neovascularization.


Eighteen clean 7-day old SD rats and 18 clean 7-day old CSNB rats were included, twelve SD rats and twelve CSNB rats were chosen randomly for oxygen induced retinopathy (OIR) modeling, and served as OIR group, six SD rats and six CSNB rats were chosen as normal control.Nine rats were chosen randomly from both SD rats and CSNB rats in OIR group, respectively.The rats were separated into 1∶1 ratio ink group, 2∶1 ratio ink group and conventional ink group, which were perfused with 1∶1 ratio ink perfusate, 2∶1 ratio ink perfusate and conventional ink, respectively.The unilateral eyes of the rats were prepared for whole-mount retina, the other eyes were performed for paraffin imbedding.The quality of retinal vascular imaging were compared among different ink perfusate groups.The normal control rats, three SD rats in OIR group and three CSNB rats in OIR group were perfused with 2∶1 ratio ink perfusate.Histopathology examination was performed on the paraffin section, and the number of nuclei breakthrough the inner limiting membrane were counted.Immunocytochemistry were performed on the paraffin section for detecting the expression of von Wilebrand factor (v-WF).


Compared with 1∶1 ratio ink perfusion and conventional ink perfusion, 2∶1 ratio ink perfusion showed the full vascular net clearly.Histopathology showed that the structure of retina was normal in the normal control group, and there were no endothelial nuclei breakthrough the inner limiting membrane.A large number of endothelial nuclei breakthrough the inner limiting membrane in the OIR group, the number of endothelial nuclei breaking through the inner limiting membrane were (23.08±2.99)/slide and (41.12±9.36)/slide for SD rats and CSNB rats, respectively.The number of endothelial nuclei breakthrough inner limiting membrane was higher in the CSNB rats than that in the SD rats, with no difference between the two group(q=1.70, P=0.50). Immunocytochemistry results showed that v-WF was positive expressed in the cells breakthrough inner limiting membrane.


Improved ink perfusion method was an easy-to-use whole-mount retina method with good repeatability.Hyperoxia can induce retinal neovascularization of CSNB rats.

Key words:

Retinal neovascularization; Congenital stationary nigh blindness; Chinese ink perfusion; Whole-mount retina

Contributor Information

Song Meixia
Department of Ophthalmology, the 153rd Central Hospital of PLA, Zhengzhou 450007, China
Li Shang
Department of Ophthalmology, the 153rd Central Hospital of PLA, Zhengzhou 450007, China
Niu Chao
Henan Eye Institute, Henan Eye Hospital, Zhengzhou 450003, China
Zhang Zuoming
Department of Clinical Aerospace Medicine, the Fourth Military Medical University, Xi’an 710032, China
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