Fig. 3: LDs are present in multiple cell types in the healthy adult mouse brain. | Nature Communications

Fig. 3: LDs are present in multiple cell types in the healthy adult mouse brain.

From: A fluorescent perilipin 2 knock-in mouse model reveals a high abundance of lipid droplets in the developing and adult brain

Fig. 3

a TdTomato expression is clearly detectable in tdTom-Plin2 brains, and Plin2 mRNA is expressed in a similar manner as in Ctrl brains. (n = 4 Ctrl and 5 tdTom-Plin2 mice, fold change +/- SEM). b Schematic illustration of a sagittal brain section, highlighting specific regions c tdTom-PLIN2 signal in the olfactory bulb, cortex, lateral ventricle well and cerebellum shows abundant LDs in the brain. Representative maximum intensity projections at low (top) and high (bottom) magnifications. (n = 3 mice, similar observations in all the mice) d TdTomato positive rings of various sizes are clearly present in the lateral ventricle wall and colocalize with the signal from staining against PLIN2, confirming that they are LDs. Representative maximum intensity projection of individual channels and a merge. e Scheme of the crossing of tdTom-Plin2 mice to Thy1GFP reporter mice, labelling neurons, and Aldh1l1GFP reporter mice, labelling astrocytes, and scheme of the FACS procedure to determine the number of GFP+ tdTomato+ cells. f FACS analysis of whole brain cell extraction from the tdTom-Plin2 mice crossed to Thy1GFP mice show 13% of all Thy1GFP positive neurons have LDs. (n = 3 mice, mean +/- SEM). g FACS analysis of whole brain cell extraction from the tdTom-Plin2 mice crossed to Aldh1l1GFP mice show that around 40% of all Aldh1l1GFP positive astrocytes have LDs. (n = 3 mice, mean +/- SEM). h Representative confocal image of an unstained hippocampal section. Thy1-GFP neurons (green) and tdTom-Plin2 signal (white), maximum projection of a z-stack over the volume of the neuron indicated. Right panel shows a 3D rendering to visualize LDs inside the indicated Thy1-GFP neurons. i Representative confocal image of an unstained hippocampal section. Aldh1l1-GFP astrocyte (green) and tdTom-Plin2 signal (white), maximum projection of a z-stack over the volume of the astrocyte). Right panel shows a 3D rendering to visualize LDs inside the indicated Aldh1l1-GFP astrocyte. j Immunohistochemical staining of the lateral wall show a large variability of endogenous tdTom-PLIN2 positive LDs in S100b-positive ependymal cells, IBA1-positive microglia, Glutamine synthase (GS)-positive astrocytes, CD31 positive endothelial cells, OLIG2-positive oligodendrocytes/oligodendrocyte progenitors and a Thy1GFP neuron. Representative maximum intensity projections of observed cell types (z-projections over the volume of the cells indicated by arrows). (n = 3 mice, similar observations in at least 10 cells per cell type and mouse). Asterisks indicate the following p-values: *** < 0.001. ns = non-significant.

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