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van Liempd, S.

Publications and source records attributed to van Liempd, S..

2 recordsLinked to original sources

Metabolic and transcriptional adaptations to phagocytosis in microglia sustain their functionality and regenerative properties

Phagocytosis of apoptotic cells, or efferocytosis, is a tightly regulated process that ensures tissue homeostasis and prevents mounting inflammatory responses. In the brain parenchyma, it is executed by microglia, which are encumbered by large numbers of apoptotic debris generated during development, in adult neurogenic niches, aging, and brain diseases. Emerging evidence suggest that phagocytosis is not limited to garbage disposal, but triggers adaptations in the phagocytes that may have a functional impact. To test it we developed an in vivo model of superphagocytosis induced by low cranial irradiation (LCI, 2Gy) that specifically induced apoptosis in the neurogenic niche of the adult hippocampus, synchronizing microglia in a phagocytic state within 6h and leading to full clearance by 24h. Single cell RNA sequencing and metabolomics revealed an unexpected oxidative stress in post-phagocytic microglia, accompanied by catabolic shutdown, mitochondrial remodeling, increased expression of galectin 3, and production of polyamines that led to cell death and compensatory proliferation. To test whether these changes impaired subsequent microglial phagocytosis, we used a glioblastoma model treated with sequential irradiation to induce tumor cell apoptosis. The phagocytosis efficiency of tumor-associated microglia/macrophages was comparable in the first and second apoptotic challenge, suggesting that the metabolic remodeling induced by phagocytosis was adaptive and destined to sustain their functionality. Finally, we assessed the functional impact of post-phagocytosis adaptations using galectin 3 deficient mice under LCI. We found that the recovery of the neurogenic niche after LCI strongly depended on galectin 3, demonstrating the regenerative capacity of post-phagocytic microglia. Overall, our data unveils the complexity of post-phagocytosis adaptations in microglia, underscoring their unexplored therapeutic potential in brain disorders.

neuroscience↗

Development and validation of a precise and accurate method to determine polyamine levels in cultured cells.

We describe a robust, fast and accurate method for the quantification of intra-cellular concentrations of the polyamines, putrescine, spermidine and spermine in cultured cells. Hydrophilic interaction liquid chromatography in combination with Time-of-Flight mass spectrometry was used to obtain high resolution data for the analytes. Assay performance was determined with respect to chromatographic resolution, quantification, analyte recovery and matrix effects. Furthermore, assay variability was determined in a biological context. Based on these variability measurements, minimal detectable effects (MDEs) which would lead to significant differences in a null-hypothesis significance test, were calculated. As such, changes in spermine could be determined with the highest sensitivity with point estimates for the MDEs of 32% between-days and 10% within-days. For spermidine, these values were 38% between-days and 16% within-days. Finally, effects for putrescine were measured least sensitive with 43% between-days and 36% within-days. Finally, we employed the method to analyze the impact of polyamine synthesis pathway inhibition and cell culture conditions which are relevant aspects for the interpretation of the biological role of polyamines.

biochemistry↗