Imaging of lactate metabolism in retinal Müller cells with a FRET nanosensor

García, Víctor Calbiague; Chen, Yiyi; Cadiz, Bárbara; Wang, Lan; Paquet-Durand, François; Schmachtenberg, Oliver

doi:10.1016/j.exer.2022.109352

Cited by 7 publications

(10 citation statements)

References 39 publications

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“…The delta ratio between depletion and saturation for Laconic was 20.3% ± 4.1 (Fig. S4), which is in line with the responses of the retina and other cells and tissues 16,32 .…”

Section: Intracellular Lactate Dynamics In the Inner Retinal Cellssupporting

confidence: 83%

“…Nonetheless, to date, there is a lack of information regarding the intracellular lactate flux in the retina. For the purpose of investigating lactate dynamics in INL cells, we expressed the FRET nanosensor h-Syn-Laconic (neurons) and GFAP-Laconic (MCs) in organotypic retinal explants, as previously described 16,32 . First, we evaluated whether nanosensor expression was functional in inner retinal neurons, by measuring the ratio between the emissions obtained from monomeric teal fluorescent protein (mTFP)/Venus in the soma of putative BCs and ACs in the INL (Fig.…”

Section: Intracellular Lactate Dynamics In the Inner Retinal Cellsmentioning

confidence: 99%

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Extracellular lactate as an alternative energy source for retinal bipolar cells

García

Chen

Cadiz

et al. 2023

Preprint

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Retinal bipolar and amacrine cells receive visual information from photoreceptors and participate in the first steps of image processing in the retina. Several studies have suggested the operation of aerobic glycolysis and a lactate shuttle system in the retina due to the high production of this metabolite under aerobic conditions. However, whether bipolar cells form part of this metabolic circuit remains unclear. Here, we show that the lactate transporter MCT2 is functionally expressed in inner retinal neurons and demonstrate their ability to consume extracellular lactate as an alternative to glucose. In rod bipolar cells, lactate is used to maintain homeostasis of various ions and electrical responses. Overall, our data contribute to a better understanding of inner retinal metabolism.

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“…The delta ratio between depletion and saturation for Laconic was 20.3% ± 4.1 (Fig. S4), which is in line with the responses of the retina and other cells and tissues 16,32 .…”

Section: Intracellular Lactate Dynamics In the Inner Retinal Cellssupporting

confidence: 83%

Section: Intracellular Lactate Dynamics In the Inner Retinal Cellsmentioning

confidence: 99%

Extracellular lactate as an alternative energy source for retinal bipolar cells

García

Chen

Cadiz

et al. 2023

Preprint

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“…To that end, we expressed the lactate nanosensor Laconic in 28 retinal explant cultures (14 animals) treated with PBM under different glucose conditions (Figure 5A). Recently, we have shown the feasibility of this assay in MCs [50]. First, we determined if PBM changes the fluorescence properties of the nanosensor.…”

Section: Effect Of Photobiomodulation Treatment On Retinal Structure ...mentioning

confidence: 99%

“…At p11 and p12, the explants were transduced by overnight incubation with 5 × 10 6 plaque-forming units (PFU) of AAV-Laconic and imaged after two weeks in culture as previously described [50]. The AAV-GFAP-Laconic was constructed by the viral vector facility of ETH Zürich (Laconic: Addgene #44238).…”

Section: Fret Measurementsmentioning

confidence: 99%

Evaluation of Photobiomodulation and Boldine as Alternative Treatment Options in Two Diabetic Retinopathy Models

García

Cadiz

Herrera

et al. 2023

IJMS

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Diabetic retinopathy causes progressive and irreversible damage to the retina through activation of inflammatory processes, overproduction of oxidative species, and glial reactivity, leading to changes in neuronal function and finally ischemia, edema, and hemorrhages. Current treatments are invasive and mostly applied at advanced stages, stressing the need for alternatives. To this end, we tested two unconventional and potentially complementary non-invasive treatment options: Photobiomodulation, the stimulation with near-infrared light, has shown promising results in ameliorating retinal pathologies and insults in several studies but remains controversial. Boldine, on the other hand, is a potent natural antioxidant and potentially useful to prevent free radical-induced oxidative stress. To establish a baseline, we first evaluated the effects of diabetic conditions on the retina with immunofluorescence, histological, and ultrastructural analysis in two diabetes model systems, obese LepRdb/db mice and organotypic retinal explants, and then tested the potential benefits of photobiomodulation and boldine treatment in vitro on retinal explants subjected to high glucose concentrations, mimicking diabetic conditions. Our results suggest that the principal subcellular structures affected by these conditions were mitochondria in the inner segment of photoreceptors, which displayed morphological changes in both model systems. In retinal explants, lactate metabolism, assayed as an indicator of mitochondrial function, was altered, and decreased photoreceptor viability was observed, presumably as a consequence of increased oxidative-nitrosative stress. The latter was reduced by boldine treatment in vitro, while photobiomodulation improved mitochondrial metabolism but was insufficient to prevent retinal structural damage caused by high glucose. These results warrant further research into alternative and complementary treatment options for diabetic retinopathy.

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“…Together with microglial cells, Müller cells are involved in the immune response through various mechanisms, including their Toll-like receptors and secretion of cytokines and chemokines (Kumar et al 2013), as well as by controlling retinal vascularisation and maintaining the bloodretinal barrier (Bringmann et al 2006). Finally, Müller cells have neuroprotective properties through the release of neurotrophic factors, the uptake and recycling of glutamate or gammaaminobutyric acid (GABA), and the secretion of the antioxidant glutathione (Bringmann et al 2006, while also supporting the metabolism of retinal neurons (Reichenbach et al 2013, Toft-Kehler et al 2016, Ola et al 2019, Calbiague Garcia et al 2023. Accordingly, selective ablation of Müller cells has been shown to result in early photoreceptor degeneration, 6 vascular abnormalities, breakdown of the BRB and neovascularisation in the retina (Shen et al 2012).…”

Section: Introductionmentioning

confidence: 99%

The glucocorticoid receptor as a master regulator of Müller cell gliosis in the diabetic retina

Pfaller,

Kaplan,

Carido

et al. 2023

Preprint

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Diabetic retinopathy (DR) is considered a primarily microvascular complication of diabetes. Müller glia cells are at the center of the retinal neurovascular unit and play a critical role in DR. We therefore investigated Müller cell-specific signaling pathways that are altered in DR to identify novel targets for gene therapy. Using a multi-omics approach on purified Müller cells from diabetic db/db mice, we found the mRNA and protein expression of the glucocorticoid receptor (GR) to be significantly decreased, while its target gene cluster was down-regulated. Further, oPOSSUM TF analysis and ATAC- sequencing identified the GR as a master regulator of Müller cell gliosis in DR. Cortisol not only increased GR phosphorylation. It also induced changes in the expression of known GR target genes in retinal explants. Finally, retinal functionality was improved by AAV-mediated overexpression of GR in Müller cells. Our study demonstrates an important role of the glial GR in DR and implies that therapeutic approaches targeting this signalling pathway should be aimed at increasing GR expression rather than the addition of more ligand.

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Imaging of lactate metabolism in retinal Müller cells with a FRET nanosensor

Cited by 7 publications

References 39 publications

Extracellular lactate as an alternative energy source for retinal bipolar cells

Extracellular lactate as an alternative energy source for retinal bipolar cells

Evaluation of Photobiomodulation and Boldine as Alternative Treatment Options in Two Diabetic Retinopathy Models

The glucocorticoid receptor as a master regulator of Müller cell gliosis in the diabetic retina

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