Utility of the DHFR-based destabilizing domain across mouse models of retinal degeneration and aging

Peng, Hui; Ramadurgum, Prerana; Woodard, DaNae R; Daniel, Steffi; Nakahara, Emi; Renwick, Marian; Aredo, Bogale; Datta, Shyamtanu; Chen, Bin; Ufret-Vincenty, Rafael; Hulleman, John D.

doi:10.1016/j.isci.2022.104206

Cited by 2 publications

(2 citation statements)

References 79 publications

(101 reference statements)

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“…However, to our knowledge, this report is the first that implicates Muller cells as playing an important role in mouse retinal recovery from a mild light injury. We have reported previously retinal recovery from light injury supported by structural and functional analyses 33 , 34 , 43 , 44 and now we are starting to understand the mechanisms behind this recovery process.…”

Section: Discussionmentioning

confidence: 87%

“…The flow chart presented in Figure 1 shows the protocols and steps used in this work starting from the application of light injury to the analysis of single cell RNA-seq (scRNA-seq). Based on our prior studies, 33 , 34 , 43 , 44 we had determined that the FCD-LIRD model led to retinal light injury that could modulated be easily by modifying the intensity of the light stimulus. In this model, maximal injury as seen on fundus photos and OCT can be seen at approximately day 5 after injury.…”

Section: Resultsmentioning

confidence: 99%

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Single Cell RNA Sequencing Analysis of Mouse Retina Identifies a Subpopulation of Muller Glia Involved in Retinal Recovery From Injury in the FCD-LIRD Model

Aredo

Kumar

Chen

et al. 2023

Invest. Ophthalmol. Vis. Sci.

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Purpose Although retinal light injury models have been useful in understanding aspects of retinal degeneration and retinal oxidative stress, information on retinal recovery from oxidative/photoinflammatory retinal injury is scarce. The fundus camera-delivered light-induced retinal degeneration model is a simple and reproducible retinal light injury model developed to recapitulate not only the retinal degeneration aspect, but also the retinal recovery from injury. In this study, we used the fundus camera-delivered light-induced retinal degeneration model to perform cell type-specific analyses of the acute and subacute retinal responses to light injury. Methods C57BL/6J eyes were collected before or after light injury (4 hours, 48 hours, and day 5). Retina samples were processed into single-cell suspensions. Droplet-based encapsulation of single cells was performed to generate libraries for sequencing. Results Gene expression analysis generated 23 clusters encompassing all known major retinal cell populations. Using unbiased analyses, we identified genes and pathways that were significantly altered in each cell type after light injury, including some cellular processes suggestive of activation of pathways for retinal recovery (e.g., synaptogenesis signaling, ephrin receptor signaling, and Reelin signaling in neurons). More importantly, our data show that a subpopulation of Muller glia cells may play an important role in the cellular recovery process. Conclusions This work identifies acute and subacute cell type-specific responses to retinal photo-oxidative injury. A subpopulation of Muller glia seems to initiate the cellular recovery process. A better understanding of these responses may be helpful in identifying therapeutic approaches to minimize retinal damage and maximize recovery after exposure to injury.

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Section: Discussionmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Single Cell RNA Sequencing Analysis of Mouse Retina Identifies a Subpopulation of Muller Glia Involved in Retinal Recovery From Injury in the FCD-LIRD Model

Aredo

Kumar

Chen

et al. 2023

Invest. Ophthalmol. Vis. Sci.

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Genetic removal of Nlrp3 protects against sporadic and R345W Efemp1-induced basal laminar deposit formation

Ortega,

Daniel,

Renwick

et al. 2024

Preprint

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Chronic, unresolved inflammation has long been speculated to serve as an initiating and propagating factor in numerous neurodegenerative diseases, including a leading cause of irreversible blindness in the elderly, age-related macular degeneration (AMD). Intracellular multiprotein complexes called inflammasomes in combination with activated caspases facilitate production of pro-inflammatory cytokines such as interleukin 1 beta. Specifically, the nucleotide-binding oligomerization (NOD)-like receptor protein 3 (NLRP3) has received heightened attention due to the wide range of stimuli to which it can respond and its potential involvement in AMD. In this study, we directly tested the role of Nlrp3 and its downstream effector, caspase 1 (Casp1) in mediating early AMD-like pathology (i.e., basal laminar deposits [BLamDs]) in wild-type (WT) mice and the Malattia Leventinese/Doyne honeycomb retinal dystrophy (ML/DHRD) mouse model (p.R345W mutation in Efemp1). Compared to aged-matched controls, R345W+/+ knockin mice demonstrated increased Muller cell gliosis, subretinal Iba-1+ microglial cells, higher Nlrp3 immunoreactivity in the retina, as well as significant transcriptional upregulation of complement component 3, Nlrp3, pro-Il1b, pro-caspase-1, and tissue inhibitor of matrix metalloproteinase 3 in the retinal pigmented epithelium (RPE)/choroid. These findings were accompanied by an age-related increase in BLamD formation in the R345W+/+ mice. Genetic elimination of either Nlrp3 or Casp1 significantly reduced both the size and coverage of BLamDs in the R345W+/+ background, highlighting an important and underappreciated pathway that could affect ML/DHRD onset and progression. Moreover, Nlrp3 knockout reduced spontaneous, idiopathic BLamDs in WT mice, suggesting translatability of our findings not only to rare inherited retinal dystrophies, but also potentially to AMD itself.

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Utility of the DHFR-based destabilizing domain across mouse models of retinal degeneration and aging

Cited by 2 publications

References 79 publications

Single Cell RNA Sequencing Analysis of Mouse Retina Identifies a Subpopulation of Muller Glia Involved in Retinal Recovery From Injury in the FCD-LIRD Model

Single Cell RNA Sequencing Analysis of Mouse Retina Identifies a Subpopulation of Muller Glia Involved in Retinal Recovery From Injury in the FCD-LIRD Model

Genetic removal of Nlrp3 protects against sporadic and R345W Efemp1-induced basal laminar deposit formation

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