An inducible Cre mouse for studying roles of the RPE in retinal physiology and disease

Choi, Elliot H.; Suh, Suk-Hwan; Einstein, David E; Leinonen, Henri; Dong, Zhiqian; Rao, Sriganesh Ramachandra; Fliesler, Steven J.; Blackshaw, Seth; Yu, Minzhong; Peachey, Neal S.; Palczewski, Krzysztof; Kiser, Philip D.

doi:10.1172/jci.insight.146604

Cited by 18 publications

(17 citation statements)

References 73 publications

(105 reference statements)

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“…This is consistent with preservation of reduced ERG expression in a subset of KCNJ13CKO ko/ko, VMD2-Cre +/+ mice. In addition, it has been shown recently that Cre expression with this model shows some leakiness with expression in other retinal cells than the RPE ( Chen et al, 2021 ; Choi et al, 2021 ). This is also apparent in Figure 5B , in which Cre expression is seen not only in the RPE but also the GCL and INL.…”

Section: Discussionmentioning

confidence: 83%

Retinal Development and Pathophysiology in Kcnj13 Knockout Mice

Jiao

Lei

et al. 2022

Front. Cell Dev. Biol.

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Purpose: We constructed and characterized knockout and conditional knockout mice for KCNJ13, encoding the inwardly rectifying K+ channel of the Kir superfamily Kir7.1, mutations in which cause both Snowflake Vitreoretinal Degeneration (SVD) and Retinitis pigmentosa (RP) to further elucidate the pathology of this disease and to develop a potential model system for gene therapy trials.Methods: A Kcnj13 knockout mouse line was constructed by inserting a gene trap cassette expressing beta-galactosidase flanked by FRT sites in intron 1 with LoxP sites flanking exon two and converted to a conditional knockout by FLP recombination followed by crossing with C57BL/6J mice having Cre driven by the VMD2 promoter. Lentiviral replacement of Kcnj13 was driven by the EF1a or VMD2 promoters.Results: Blue-Gal expression is evident in E12.5 brain ventricular choroid plexus, lens, neural retina layer, and anterior RPE. In the adult eye expression is seen in the ciliary body, RPE and choroid. Adult conditional Kcnj13 ko mice show loss of photoreceptors in the outer nuclear layer, inner nuclear layer thinning with loss of bipolar cells, and thinning and disruption of the outer plexiform layer, correlating with Cre expression in the overlying RPE which, although preserved, shows morphological disruption. Fundoscopy and OCT show signs of retinal degeneration consistent with the histology, and photopic and scotopic ERGs are decreased in amplitude or extinguished. Lentiviral based replacement of Kcnj13 resulted in increased ERG c- but not a- or b- wave amplitudes.Conclusion: Ocular KCNJ13 expression starts in the choroid, lens, ciliary body, and anterior retina, while later expression centers on the RPE with no/lower expression in the neuroretina. Although KCNJ13 expression is not required for survival of the RPE, it is necessary for RPE maintenance of the photoreceptors, and loss of the photoreceptor, outer plexiform, and outer nuclear layers occur in adult KCNJ13 cKO mice, concomitant with decreased amplitude and eventual extinguishing of the ERG and signs of retinitis pigmentosa on fundoscopy and OCT. Kcnj13 replacement resulting in recovery of the ERG c- but not a- and b-waves is consistent with the degree of photoreceptor degeneration seen on histology.

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Section: Discussionmentioning

confidence: 83%

Retinal Development and Pathophysiology in Kcnj13 Knockout Mice

Jiao

Lei

et al. 2022

Front. Cell Dev. Biol.

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“…We generated a new Rpe65 CreERT2 Cre driver line with a genomic design resembling a previously published model [ 50 ]. The Rpe65 CreERT/ + 2 mice showed normal retinal function and morphology, as assessed by ERG and histological sections, respectively (Additional file 2 : Fig.…”

Section: Resultsmentioning

confidence: 99%

“…S2C, D). Previously, Choi et al [ 50 ] demonstrated that the Rpe65 CreERT2 homozygous mice exhibited reduced RPE65 isomerase levels compared to heterozygous and WT mice. However, we did not observe a decrease in RPE65 expression in our mice (Additional file 2 : Fig.…”

Section: Resultsmentioning

confidence: 99%

Targeting miR-181a/b in retinitis pigmentosa: implications for disease progression and therapy

Costa,

Quinn,

et al. 2024

Cell Biosci

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Background Retinitis pigmentosa (RP) is a genetically heterogeneous group of degenerative disorders causing progressive vision loss due to photoreceptor death. RP affects other retinal cells, including the retinal pigment epithelium (RPE). MicroRNAs (miRs) are implicated in RP pathogenesis, and downregulating miR-181a/b has shown therapeutic benefit in RP mouse models by improving mitochondrial function. This study investigates the expression profile of miR-181a/b in RPE cells and the neural retina during RP disease progression. We also evaluate how miR-181a/b downregulation, by knocking out miR-181a/b-1 cluster in RPE cells, confers therapeutic efficacy in an RP mouse model and explore the mechanisms underlying this process. Results Our findings reveal distinct expression profiles, with downregulated miR-181a/b in RPE cells suggesting a protective response and upregulated miR-181a/b in the neural retina indicating a role in disease progression. We found that miR-181a/b-2, encoded in a separate genomic cluster, compensates for miR-181a/b-1 ablation in RPE cells at late time points. The transient downregulation of miR-181a/b in RPE cells at post-natal week 6 (PW6) led to improved RPE morphology, retarded photoreceptor degeneration and decreased RPE aerobic glycolysis. Conclusions Our study elucidates the underlying mechanisms associated with the therapeutic modulation of miR-181a/b, providing insights into the metabolic processes linked to its RPE-specific downregulation. Our data further highlights the impact of compensatory regulation between miR clusters with implications for the development of miR-based therapeutics. Graphical Abstract

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“…ADAM17 is a member of the adamalysin family that are responsible for the shedding (ectodomain cleavage) of cell membrane proteins. ADAM17 expression has been shown in endothelial cells and different cell types of the CNS [ 24 , 45 ]. In mammals, the forebrain, where optic vesicle formation is ADAM17-positive, suggests the necessity of ADAM17 for retinal morphogenesis [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

“…A day before ERG testing, the mice were dark-adapted for > 12 h. Animal handling was performed under dim red light. Anesthetized mice were positioned on the thermal pad (37 ± 1 °C), and the stimulator was placed on the surface of the cornea with the largest contact surface, according to the protocol suggested by the instructor [ 24 ]. The dark-adapted ERG and OPs were stimulated with an ascending flash series (intensity 0.01, 0.1 and 1 cd.s/m 2 ; frequency 1 Hz), and the signal waves were acquired from 50 ms pre-trigger to 300 ms post-trigger.…”

Section: Methodsmentioning

confidence: 99%

Blue light exposure collapses the inner blood-retinal barrier by accelerating endothelial CLDN5 degradation through the disturbance of GNAZ and the activation of ADAM17

Chan

Hsiao

Wan

et al. 2023

Fluids Barriers CNS

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Blue light is part of the natural light spectrum that emits high energy. Currently, people are frequently exposed to blue light from 3C devices, resulting in a growing incidence of retinopathy. The retinal vasculature is complex, and retinal vessels not only serve the metabolic needs of the retinal sublayers, but also maintain electrolyte homeostasis by forming the inner blood-retinal barrier (iBRB). The iBRB, which is primarily composed of endothelial cells, has well-developed tight junctions. However, with exposure to blue light, the risks of targeting retinal endothelial cells are currently unknown. We found that endothelial claudin-5 (CLDN5) was rapidly degraded under blue light, coinciding with the activation of a disintegrin and metalloprotease 17 (ADAM17), even at non-cytotoxic lighting. An apparently broken tight junction and a permeable paracellular cleft were observed. Mice exposed to blue light displayed iBRB leakage, conferring attenuation of the electroretinogram b-wave and oscillatory potentials. Both pharmacological and genetic inhibition of ADAM17 remarkably alleviated CLDN5 degradation induced by blue light. Under untreated condition, ADAM17 is sequestered by GNAZ (a circadian-responsive, retina-enriched inhibitory G protein), whereas ADAM17 escapes from GNAZ by blue light illuminance. GNAZ knockdown led to ADAM17 hyperactivation, CLDN5 downregulation, and paracellular permeability in vitro, and retinal damage mimicked blue light exposure in vivo. These data demonstrate that blue light exposure might impair the iBRB by accelerating CLDN5 degradation through the disturbance of the GNAZ-ADAM17 axis.

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An inducible Cre mouse for studying roles of the RPE in retinal physiology and disease

Cited by 18 publications

References 73 publications

Retinal Development and Pathophysiology in Kcnj13 Knockout Mice

Retinal Development and Pathophysiology in Kcnj13 Knockout Mice

Targeting miR-181a/b in retinitis pigmentosa: implications for disease progression and therapy

Blue light exposure collapses the inner blood-retinal barrier by accelerating endothelial CLDN5 degradation through the disturbance of GNAZ and the activation of ADAM17

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