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Peroxisomal ether-GPs synthesis was disrupted after TBI. A: schematic diagram showing peroxisomal ether-GPs synthesizing steps. It is initiated by the acylation of dihydroxyacetone phosphate (DHAP) <t>by</t> <t>GNPAT</t> that generates 1-acyl-DHAP which is then converted to 1-O-alkyl-DHAP by <t>AGPS.</t> 1-O-alkyl-DHAP is then converted to 1-O-alkylglycerol phosphate which is transported to the endoplasmic reticulum for the generation of fully formed ether-GPs. B: Western blots of peroxisomal enzymes GNPAT and AGPS in the cortical tissue lysates of sham and TBI mice. Data presented as mean ± SEM. n = 4; ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. C: Western blots and (D) corresponding quantification of peroxisomal enzymes GNPAT and AGPS in the peroxisomal and cytosolic fractions prepared from the sham and TBI (PID 1) mouse cortices. ABCD3 (PMP70) and PEX14 are markers of peroxisomal membrane and α-tubulin is of cytosolic fraction. Data presented as mean ± SEM. n = 4. ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. E: Western blots and corresponding (F) quantification of GNPAT and AGPS in the peroxisomal (ABCD3+) and cytosolic (α-tubulin+) fractions of sham and TBI mouse cortices (PID 28). Data = Mean ± SEM; n = 5. ∗∗ P < 0.01, Students' t test. G: 60X images of AGPS and peroxisomal membrane marker PEX14. H: Quantification of punctate versus diffused AGPS ratio in sham (blue) and injured (1 day after TBI) (red) brain sections. Data presented as mean ± SEM. n = 3; ∗ P < 0.05; Students' t test.
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Peroxisomal ether-GPs synthesis was disrupted after TBI. A: schematic diagram showing peroxisomal ether-GPs synthesizing steps. It is initiated by the acylation of dihydroxyacetone phosphate (DHAP) <t>by</t> <t>GNPAT</t> that generates 1-acyl-DHAP which is then converted to 1-O-alkyl-DHAP by <t>AGPS.</t> 1-O-alkyl-DHAP is then converted to 1-O-alkylglycerol phosphate which is transported to the endoplasmic reticulum for the generation of fully formed ether-GPs. B: Western blots of peroxisomal enzymes GNPAT and AGPS in the cortical tissue lysates of sham and TBI mice. Data presented as mean ± SEM. n = 4; ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. C: Western blots and (D) corresponding quantification of peroxisomal enzymes GNPAT and AGPS in the peroxisomal and cytosolic fractions prepared from the sham and TBI (PID 1) mouse cortices. ABCD3 (PMP70) and PEX14 are markers of peroxisomal membrane and α-tubulin is of cytosolic fraction. Data presented as mean ± SEM. n = 4. ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. E: Western blots and corresponding (F) quantification of GNPAT and AGPS in the peroxisomal (ABCD3+) and cytosolic (α-tubulin+) fractions of sham and TBI mouse cortices (PID 28). Data = Mean ± SEM; n = 5. ∗∗ P < 0.01, Students' t test. G: 60X images of AGPS and peroxisomal membrane marker PEX14. H: Quantification of punctate versus diffused AGPS ratio in sham (blue) and injured (1 day after TBI) (red) brain sections. Data presented as mean ± SEM. n = 3; ∗ P < 0.05; Students' t test.
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Peroxisomal ether-GPs synthesis was disrupted after TBI. A: schematic diagram showing peroxisomal ether-GPs synthesizing steps. It is initiated by the acylation of dihydroxyacetone phosphate (DHAP) by GNPAT that generates 1-acyl-DHAP which is then converted to 1-O-alkyl-DHAP by AGPS. 1-O-alkyl-DHAP is then converted to 1-O-alkylglycerol phosphate which is transported to the endoplasmic reticulum for the generation of fully formed ether-GPs. B: Western blots of peroxisomal enzymes GNPAT and AGPS in the cortical tissue lysates of sham and TBI mice. Data presented as mean ± SEM. n = 4; ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. C: Western blots and (D) corresponding quantification of peroxisomal enzymes GNPAT and AGPS in the peroxisomal and cytosolic fractions prepared from the sham and TBI (PID 1) mouse cortices. ABCD3 (PMP70) and PEX14 are markers of peroxisomal membrane and α-tubulin is of cytosolic fraction. Data presented as mean ± SEM. n = 4. ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. E: Western blots and corresponding (F) quantification of GNPAT and AGPS in the peroxisomal (ABCD3+) and cytosolic (α-tubulin+) fractions of sham and TBI mouse cortices (PID 28). Data = Mean ± SEM; n = 5. ∗∗ P < 0.01, Students' t test. G: 60X images of AGPS and peroxisomal membrane marker PEX14. H: Quantification of punctate versus diffused AGPS ratio in sham (blue) and injured (1 day after TBI) (red) brain sections. Data presented as mean ± SEM. n = 3; ∗ P < 0.05; Students' t test.

Journal: Journal of Lipid Research

Article Title: Peroxisomal ether-glycerophospholipid synthesis is dysregulated after TBI

doi: 10.1016/j.jlr.2025.100821

Figure Lengend Snippet: Peroxisomal ether-GPs synthesis was disrupted after TBI. A: schematic diagram showing peroxisomal ether-GPs synthesizing steps. It is initiated by the acylation of dihydroxyacetone phosphate (DHAP) by GNPAT that generates 1-acyl-DHAP which is then converted to 1-O-alkyl-DHAP by AGPS. 1-O-alkyl-DHAP is then converted to 1-O-alkylglycerol phosphate which is transported to the endoplasmic reticulum for the generation of fully formed ether-GPs. B: Western blots of peroxisomal enzymes GNPAT and AGPS in the cortical tissue lysates of sham and TBI mice. Data presented as mean ± SEM. n = 4; ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. C: Western blots and (D) corresponding quantification of peroxisomal enzymes GNPAT and AGPS in the peroxisomal and cytosolic fractions prepared from the sham and TBI (PID 1) mouse cortices. ABCD3 (PMP70) and PEX14 are markers of peroxisomal membrane and α-tubulin is of cytosolic fraction. Data presented as mean ± SEM. n = 4. ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. E: Western blots and corresponding (F) quantification of GNPAT and AGPS in the peroxisomal (ABCD3+) and cytosolic (α-tubulin+) fractions of sham and TBI mouse cortices (PID 28). Data = Mean ± SEM; n = 5. ∗∗ P < 0.01, Students' t test. G: 60X images of AGPS and peroxisomal membrane marker PEX14. H: Quantification of punctate versus diffused AGPS ratio in sham (blue) and injured (1 day after TBI) (red) brain sections. Data presented as mean ± SEM. n = 3; ∗ P < 0.05; Students' t test.

Article Snippet: Primary antibodies: AGPS (1:1000; Thermo Scientific, PA5-87935), GNPAT (1:1000; Thermo Scientific, PA5-36447), ABCD3/PMP70 (1:1000, Thermo Scientific, PA1-650), α-Tubulin (1:500; AA4.3-s, developed by Walsh, C. and obtained from Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52,242), β-actin/ACTB (1:10,000; Sigma, A1978) and PEX7 (1:1,000, Proteintech, 20614-1-AP) and PEX14 (1:1000, Proteintech, 10594-1-AP).

Techniques: Western Blot, Membrane, Marker