Histone Deacetylases Contribute to Excitotoxicity-Triggered Degeneration of Retinal Ganglion Cells In Vivo

Schlüter, Annabelle; Aksan, Bahar; Fioravanti, Rossella; Mai, Antonello; Mauceri, Daniela

doi:10.1007/s12035-019-01658-x

Cited by 25 publications

(21 citation statements)

References 76 publications

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“…Double and triple immunofluorescence was performed on free-floating whole retinae as described previously (Schlüter et al, 2019). Retinae were blocked for 4-5 h (in 0.5% Triton X-100, 0.2% bovine serum albumin and 0.02% sodium azide in 1x PBS) at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Dynamic Regulation of Synaptopodin and the Axon Initial Segment in Retinal Ganglion Cells During Postnatal Development

Schlüter

Roßberger

Dannehl

et al. 2019

Front. Cell. Neurosci.

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A key component allowing a neuron to function properly within its dynamic environment is the axon initial segment (AIS), the site of action potential generation. In visual cortex, AIS of pyramidal neurons undergo periods of activity-dependent structural plasticity during development. However, it remains unknown how AIS morphology is organized during development for downstream cells in the visual pathway (retinal ganglion cells; RGCs) and whether AIS retain the ability to dynamically adjust to changes in network state. Here, we investigated the maturation of AIS in RGCs during mouse retinal development, and tested putative activity-dependent mechanisms by applying visual deprivation with a focus on the AIS-specific cisternal organelle (CO), a presumed Ca 2+ -store. Whole-mount retinae from wildtype and Thy1-GFP transgenic mice were processed for multi-channel immunofluorescence using antibodies against AIS scaffolding proteins ankyrin-G, βIV-spectrin and the CO marker synaptopodin (synpo). Confocal microscopy in combination with morphometrical analysis of AIS length and position as well as synpo cluster size was performed. Data indicated that a subset of RGC AIS contains synpo clusters and that these show significant dynamic regulation in size during development as well as after visual deprivation. Using super resolution microscopy, we addressed the subcellular localization of synpo in RGC axons. Similar to cortical neurons, RGCs show a periodic distribution of AIS scaffolding proteins. A previously reported scaffold-deficient nanodomain correlating with synpo localization is not evident in all RGC AIS. In summary, our work demonstrates a dynamic regulation of both the AIS and synpo in RGCs during retinal development and after visual deprivation, providing first evidence that the AIS and CO in RGCs can undergo structural plasticity in response to changes in network activity.

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Section: Methodsmentioning

confidence: 99%

Dynamic Regulation of Synaptopodin and the Axon Initial Segment in Retinal Ganglion Cells During Postnatal Development

Schlüter

Roßberger

Dannehl

et al. 2019

Front. Cell. Neurosci.

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“…It is also noteworthy mentioning that, as far as we are aware, ours is the first study reporting a protein-protein interaction between DAPK1 and several histones, and that these physical interactions weaken after OGD. This is interesting since it has been described that histones may play a role in orchestrating the neuronal transcriptional response to experimental stroke by MCAO, OGD [29][30][31], and other challenges to cell survival such as excitotoxicity or ferroptosis [32][33][34]. Notably, we have also found that peroxiredoxin (PRDX) 1, PRDX2 and PRDX3, members of a family of antioxidant enzymes of crucial physiological importance, interact with DAPK1 and that this degree of association decreases after OGD.…”

Section: Discussionmentioning

confidence: 62%

Comparative Proteomics Unveils LRRFIP1 as a New Player in the DAPK1 Interactome of Neurons Exposed to Oxygen and Glucose Deprivation

DeGregorio-Rocasolano

Guirao

Ponce

et al. 2020

Antioxidants

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Death-associated protein kinase 1 (DAPK1) is a pleiotropic hub of a number of networked distributed intracellular processes. Among them, DAPK1 is known to interact with the excitotoxicity driver NMDA receptor (NMDAR), and in sudden pathophysiological conditions of the brain, e.g., stroke, several lines of evidence link DAPK1 with the transduction of glutamate-induced events that determine neuronal fate. In turn, DAPK1 expression and activity are known to be affected by the redox status of the cell. To delineate specific and differential neuronal DAPK1 interactors in stroke-like conditions in vitro, we exposed primary cultures of rat cortical neurons to oxygen/glucose deprivation (OGD), a condition that increases reactive oxygen species (ROS) and lipid peroxides. OGD or control samples were co-immunoprecipitated separately, trypsin-digested, and proteins in the interactome identified by high-resolution LC-MS/MS. Data were processed and curated using bioinformatics tools. OGD increased total DAPK1 protein levels, cleavage into shorter isoforms, and dephosphorylation to render the active DAPK1 form. The DAPK1 interactome comprises some 600 proteins, mostly involving binding, catalytic and structural molecular functions. OGD up-regulated 190 and down-regulated 192 candidate DAPK1-interacting proteins. Some differentially up-regulated interactors related to NMDAR were validated by WB. In addition, a novel differential DAPK1 partner, LRRFIP1, was further confirmed by reverse Co-IP. Furthermore, LRRFIP1 levels were increased by pro-oxidant conditions such as ODG or the ferroptosis inducer erastin. The present study identifies novel partners of DAPK1, such as LRRFIP1, which are suitable as targets for neuroprotection.

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“…[16][17][18][19][20] Indeed, high concentrations of glutamate or NMDA delivered via intravitreal injections can be utilized as a tool to cause RGC death in vivo. [21][22][23][24][25][26] We performed a single intravitreal injection of 10 nmol of NMDA in adult mice and analyzed NMDA-induced RGC death at two different time points (day 1 [d1] and day 7 [d7]) (Figure 1). Brn3a served as a marker to identify and quantify RGCs 27 in central and peripheral regions of retinal whole mounts ( Figures 1A and 1B).…”

Section: Resultsmentioning

confidence: 99%

“…32,40,65,73 Excitotoxicity in the retina alters the subcellular distribution, and thus activity, of the epigenetic regulators histone deacetylases (HDACs) in RGCs. 26 The activity of one particular HDAC, namely HDAC4, regulates the expression of VEGFD. 51 It remains an open question whether such epigenetic regulators are involved in the decrease of VEGFD levels described in the present study.…”

Section: Discussionmentioning

confidence: 99%

“…Somatic levels of VEGFD in sagittal retinal sections were analyzed as previously published. 26 Fluorescent signals were measured in cells located in the ganglion cell layer by manually drawing regions of interest around the ganglion cell layer in four to five images, each corresponding to one individual 16-mm sagittal retina section using Im-ageJ/Fiji. For each eye/condition, mean integrated densities were calculated out of the single fluorescence values within each sagittal retinal section.…”

Section: Analysis Of Vegfd Expressionmentioning

confidence: 99%

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VEGFD Protects Retinal Ganglion Cells and, consequently, Capillaries against Excitotoxic Injury

Schlüter

Aksan

Diem

et al. 2020

Molecular Therapy - Methods & Clinical Development

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In the central nervous system, neurons and the vasculature influence each other. While it is well described that a functional vascular system is trophic to neurons and that vascular damage contributes to neurodegeneration, the opposite scenario in which neural damage might impact the microvasculature is less defined. In this study, using an in vivo excitotoxic approach in adult mice as a tool to cause specific damage to retinal ganglion cells, we detected subsequent damage to endothelial cells in retinal capillaries. Furthermore, we detected decreased expression of vascular endothelial growth factor D (VEGFD) in retinal ganglion cells. In vivo VEGFD supplementation via neuronal-specific viral-mediated expression or acute intravitreal delivery of the mature protein preserved the structural and functional integrity of retinal ganglion cells against excitotoxicity and, additionally, spared endothelial cells from degeneration. Viral-mediated suppression of expression of the VEGFD-binding receptor VEGFR3 in retinal ganglion cells revealed that VEGFD exerts its protective capacity directly on retinal ganglion cells, while protection of endothelial cells is the result of upheld neuronal integrity. These findings suggest that VEGFD supplementation might be a novel, clinically applicable approach for neuronal and vascular protection.

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Histone Deacetylases Contribute to Excitotoxicity-Triggered Degeneration of Retinal Ganglion Cells In Vivo

Cited by 25 publications

References 76 publications

Dynamic Regulation of Synaptopodin and the Axon Initial Segment in Retinal Ganglion Cells During Postnatal Development

Dynamic Regulation of Synaptopodin and the Axon Initial Segment in Retinal Ganglion Cells During Postnatal Development

Comparative Proteomics Unveils LRRFIP1 as a New Player in the DAPK1 Interactome of Neurons Exposed to Oxygen and Glucose Deprivation

VEGFD Protects Retinal Ganglion Cells and, consequently, Capillaries against Excitotoxic Injury

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