Rs1h−/y exon 3-del rat model of X-linked retinoschisis with early onset and rapid phenotype is rescued by RS1 supplementation
Authors: Maria Mercedes Campos
Yichao Li
Haohua Qian
Camasamudram Vijayasarathy
Paul A Sieving
Yong Zeng
Journal: Gene Therapy (2021)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Animal models; X-linked juvenile retinoschisis; XLRS; retinoschisin; RS1 protein
Affiliations: National Eye Institute, National Institutes of Health, Bethesda, MD, USA., National Eye Institute, National Institutes of Health, Bethesda, MD, USA. .

Abstract:  Animal models of X-linked juvenile retinoschisis (XLRS) are valuable tools for understanding basic biochemical function of retinoschisin (RS1) protein and to investigate outcomes of preclinical efficacy and toxicity studies. In order to work with an eye larger than mouse, we generated and characterized an Rs1h-/y knockout rat model created by removing exon 3. This rat model expresses no normal RS1 protein. The model shares features of an early onset and more severe phenotype of human XLRS. The morphologic pathology includes schisis cavities at postnatal day 15 (p15), photoreceptors that are misplaced into the subretinal space and OPL, and a reduction of photoreceptor cell numbers by p21. By 6 mo age only 1-3 rows of photoreceptors nuclei remain, and the inner/outer segment layers and the OPL shows major changes. Electroretinogram recordings show functional loss with considerable reduction of both the a-wave and b-wave by p28, indicating early age loss and dysfunction of photoreceptors. The ratio of b-/a-wave amplitudes indicates impaired synaptic transmission to bipolar cells in addition. Supplementing the Rs1h-/y exon3-del retina with normal human RS1 protein using AAV8-RS1 delivery improved the retinal structure. This Rs1h-/y rat model provides a further tool to explore underlying mechanisms of XLRS pathology and to evaluate therapeutic intervention for the XLRS condition. 

Retracted: No

PubPeer comments:
 Figure 1B: Do the authors have a better quality image of the gel available? Preferably, where the wells are visible? It can be pasted below. The migration of the DNA ladder seems a bit strange, under normal conditions, on an agarose gel, the 2000 bp band should run closer to the 3000 bp marker, in this case it s nearer the 800 bp marker. I also think it s fairly unusual for the 100 bp marker to have the same intensity as the 3000 bp marker without any additional diffusion. There aren t any methods describing how the agarose gel was run, in the main text, or any supplementary methods. The migration in Figure 5A looks more  natural (Admittedly subjective!). Since this was only published last year, hopefully the original data is close to hand, and I m sure that will satisfy my curiosity. Thanks! ![file] , 
 #1 Thanks for the comment. The electrophoresis to separate the PCR product used is Flushing gel system made by Lonza, is not regular agarose gel. please checking their products. Yong , 
 #2 Hi Yong Zeng, thank you for your prompt response, I presume you re referring to the FlashGelu2122 System? I had a look at their [product literature] (although I can t help but feel that it might have been more appropriate for you to do that). It looks like you have labelled your ladder wrong. The 2000 bp should be 1500 bp, which would make the position of the band in your gel more logical. My initial concern was that your gel had been manipulated, or possibly labelled upside down, if you felt that the image was sufficiently important to include in the manuscript, I think it s fair to assume that the methods should be documented, the gel correctly labelled, and the raw data made available when requested. ![file] ,