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Figure 7 | BMC Pharmacology

Figure 7

From: Lipid phosphate phosphatase inhibitors locally amplify lysophosphatidic acid LPA1 receptor signalling in rat brain cryosections without affecting global LPA degradation

Figure 7

Rat cerebellar membranes principally metabolize LPA via the LPA → MAG → G pathway. (a) Rat cerebellar membranes (1 μg/well) were incubated in the absence (basal) or presence of LPA, glycerol 3-phosphate (GP), or the phosphatase-resistant LPA analog (2 S)-OMPT (10 μM final concentration each). Pi generation was kinetically monitored for 90 min using a fluorescent assay as described in Methods. Rat cerebellar membranes generate Pi from exogenous LPA but not from exogenous GP or (2 S)-OMPT. The data are expressed as nmol of Pi generated per mg of protein during the 90 min incubation (mean + SEM, n = 4 for basal and LPA, n = 3 for GP and (2 S)-OMPT). Significance level: ***p < 0.001 compared to basal. (b) Rat cerebellar membranes were pretreated for 30 min with DMSO or the broadly-acting serine hydrolase inhibitor MAFP (1 μM) or the MGL-specific inhibitor JZL184 (100 μM). This was followed by 90 min incubation in the absence (basal) or presence of LPA (10 μM final concentration). Glycerol generation was kinetically monitored for 90 min using a fluorescent assay as described in Methods. Rat cerebellar membranes readily generate glycerol from exogenous LPA; this response is largely blocked by MAFP and partially so by JZL184. The data are expressed as nmol of glycerol generated per mg of protein during the 90 min incubation (mean + SEM, n = 4). Significance level: ***p < 0.001 compared to either basal or particular treatment. (c) Simultaneous kinetic monitoring of Pi and glycerol generation from exogenous LPA by rat cerebellar membranes during 90 min incubation. Note that Pi generation precedes and exceeds that of glycerol during the early time-points (10–30 min) suggesting temporal first phosphatase (LPA → MAG) then lipase (MAG → G) action. The data are expressed as nmol of Pi or glycerol generated per mg of protein and are means of duplicate wells from one representative experiment.

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