The activation of GPR27 increases cytosolic L-lactate in 3T3 embryonic cells and astrocytes

Dolanc, Dorian (author), Zorec, Tomaž Mark (author), Smole, Zala (author), Maver, Anja (author), Horvat, Anemari (author), Pillaiyar, Thanigaimalai (author), Trkov, Saša (author), Vardjan, Nina (author), Kreft, Marko (author), Haque Chowdhury, Helena (author), Zorec, Robert (author)

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.

• Language: English
• Keywords: 3T3 embryonic cells, astrocytes, aerobic glycolysis, cytosolic L-lactate, FRET nanosensor, G-protein-coupled receptors, GPR27, agonists
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine; BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2022
• Number of pages: 14 str.
• Numbering: Vol. 11, iss. 6, art. 1009
• PID: 20.500.12556/RUL-137536
• UDC: 616
• ISSN on article: 2073-4409
• DOI: 10.3390/cells11061009
• COBISS.SI-ID: 101228291
• Publication date in RUL: 21.06.2022

Record is a part of a journal

• Title: Cells
• Shortened title: Cells
• Publisher: MDPI
• ISSN: 2073-4409
• COBISS.SI-ID: 519958809

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
• Licensing start date: 16.03.2022

Secondary language

• Language: Slovenian
• Keywords: 3T3 embrionalne celice, astrociti, aerobna glikoliza