20.500.12556/RUL-137536 The activation of GPR27 increases cytosolic L-lactate in 3T3 embryonic cells and astrocytes G-protein-coupled receptors (GPCRs) represent a family with over 800 members in humans, and one-third of these are targets for approved drugs. A large number of GPCRs have unknown physiologic roles. Here, we investigated GPR27, an orphan GPCR belonging to the family of super conserved receptor expressed in the brain, with unknown functions. Cytosolic levels of L-lactate ([lactate]$_i$), the end product of aerobic glycolysis, were measured with the Laconic fluorescence resonance energy transfer nanosensor. In single 3T3 wild-type (WT) embryonic cells, the application of 8535 (1 µM), a surrogate agonist known to activate GPR27, resulted in an increase in [lactate]$_i$. Similarly, an increase was recorded in primary rat astrocytes, a type of neuroglial cell abundant in the brain, which contain glycogen and express enzymes of aerobic glycolysis. In CRISPR-Cas9 GPR27 knocked out 3T3 cells, the 8535-induced increase in [lactate]$_i$ was reduced compared with WT controls. Transfection of the GPR27-carrying plasmid into the 3T3KOGPR27 cells rescued the 8535-induced increase in [lactate]$_i$. These results indicate that stimulation of GPR27 enhances aerobic glycolysis and L-lactate production in 3T3 cells and astrocytes. Interestingly, in the absence of GPR27 in 3T3 cells, resting [lactate]$_i$ was increased in comparison with controls, further supporting the view that GPR27 regulates L-lactate homeostasis. 3T3 embryonic cells astrocytes aerobic glycolysis cytosolic L-lactate FRET nanosensor G-protein-coupled receptors GPR27 agonists 3T3 embrionalne celice astrociti aerobna glikoliza true false true Angleški jezik Slovenski jezik Članek v reviji 2022-06-21 10:49:04 2022-06-21 10:49:09 2022-09-15 04:15:21 0000-00-00 00:00:00 2022 0 0 14 str. iss. 6, art. 1009 Vol. 11 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616 2073-4409 10.3390/cells11061009 101228291 cells-11-01009-v3.pdf cells-11-01009-v3.pdf 1 ECCBED06C58869521865A07DA87945C2 8aa91a9aa0ed07abd5b762b5c48bbe75a6719021f068efd161797a3868281c75 2d85ec00-f13f-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=157519 https://www.mdpi.com/2073-4409/11/6/1009 0 eca4eedc-f13e-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=157518 Medicinska fakulteta Biotehniška fakulteta 0 0 0