Abstract
The optic nerve transfers visual information from the retina to the brain through the axons of retinal ganglion cells (RGCs). In adult mammals, optic nerve injuries and progressive degenerative diseases lead to the irreversible loss of RGCs, resulting in vision loss and blindness. Optogenetic models have proved useful in manipulating the growth of RGCs through expression and stimulation of channelrhodopsins (Chr2) in RGCs using the RGC-specific thy-1 promoter. Using transgenic Chr2 mouse (Thy1-ChR2-EYFP) as a model of regeneration, we profile the lipid changes which occur after traumatic optic nerve crush, light stimulation and forced RGC axonal growth. Thy1-ChR2-EYFP and control (C57BL/6) mice were divided in four groups each – 1) no crush and no stimulation, 2) no crush with stimulation, 3) crush and without stimulation, and 4) crush with stimulation. After euthanasia, the optic nerves were collected for lipidomic analysis. The Bligh and Dyer method was used for lipid extraction, followed by mass spectrometry lipid profiling with a Q-Exactive Orbitrap Liquid Chromatography-Mass Spectrometer (LC MS-MS). The raw scans were analysed with LipidSearch 4.1.3 and the statistical analysis was conducted through Metaboanalyst 4.0. This data is available at Metabolomics Workbench, study ID ST001381: [https://www.metabolomicsworkbench.org/data/DRCCMetadata.php?Mode=Study&StudyID=ST001381&StudyType=MS&ResultType=5].
Highlights
The optic nerve transfers visual information from the retina to the brain through the axons of retinal ganglion cells (RGCs)
Using transgenic Chr2 mouse (Thy1-ChR2-EYFP) as a model of regeneration, we profile the lipid changes which occur after traumatic optic nerve crush, light stimulation and forced RGC axonal growth
The data provides insights into the lipid changes that occur after traumatic optic nerve injury and axonal regeneration in two mice models - Thy1-ChR2-EYFP mice and C57BL/6 J control mice
Summary
Mass-Spectrometric lipidomics analysis was performed on optic nerves from the transgenic channelrhodopsin 2 mouse (Thy1-ChR2-EYFP) as a model for stimulated axonal regeneration and from control C57BL/6 J mouse after optic nerve crush and light stimulation to activate cation channels through ChR2 to stimulate RGCs. Optogenetics employs genetic and optical methods for the precise stimulation of specific cell types which express ChR2 [1]. The Thy1-ChR2EYFP mouse expresses channelrhodopsin, under the thy-1 RGC specific promoter, in retinal ganglion cells (RGCs) which are targeted for stimulation by ChR2 cation channel acti-. The raw scans from a Q-Exactive Orbitrap Mass Spectrometer were analysed with Lipidsearch 4.1.3. All raw data were uploaded and processed with Lipidsearch 4.1.3. Partial least square discriminant analyses of the lipid profiling data (Fig. 2) was obtained with Metaboanalyst 4.0, as well at the lipid species heatmap (Fig. 3)
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