A 2A R mediated modulation in IP 3 levels altering the [Ca 2+ ] i through cAMP-dependent PKA signalling pathway

Stimulation of A receptors (A R) coupled to G /olf protein activates Adenylyl cyclase (AC) leading to the release of cAMP which activates the cAMP-dependent PKA phosphorylation. The possible role of A R in the modulation of free cytosolic Ca concentration ([Ca ] ) involving IP , cAMP and PKA was inv...

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Bibliographic Details
Published in:Biochimica et biophysica acta. General subjects 2022-12, Vol.1866 (12), p.130242
Main Authors: Barodia, Sandeep Kumar, Sophronea, Tuithung, Luthra, Pratibha Mehta
Format: Article
Language:English
Subjects:
Adenosine-5'-(N-ethylcarboxamide) - pharmacology
Adenylyl Cyclases - metabolism
Adenylyl Cyclases - pharmacology
Cyclic AMP - metabolism
HEK293 Cells
Humans
Signal Transduction
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Summary:Stimulation of A receptors (A R) coupled to G /olf protein activates Adenylyl cyclase (AC) leading to the release of cAMP which activates the cAMP-dependent PKA phosphorylation. The possible role of A R in the modulation of free cytosolic Ca concentration ([Ca ] ) involving IP , cAMP and PKA was investigated in HEK 293-A R. The levels of IP and cAMP were observed by enzyme immunoassay detection method and [Ca ] using Fluo-4 AM. Moreover, cAMP-dependent PKA was determined using the PKA Colorimetric Activity Kit. We observed that the cells pre-treated with A R agonist NECA showed increased levels of cAMP, PKA, IP and [Ca ] levels. However, the reverse effect was observed with A R antagonists (ZM241385 and caffeine). Blocking the G /PLC/DAG/IP pathway with neomycin, a PLC inhibitor did not affect the modulation of IP and [Ca ] levels in HEK 293-A R cells. To investigate the G /AC/cAMP/PKA, HEK 293-A R cells pre-treated with pertussis toxin followed by forskolin in the presence of A R agonist (NECA) showed no effect on cAMP levels. Further, G /AC/cAMP/PKA pathway was investigated to elucidate the role of cAMP-dependent PKA in IP mediated [Ca ] modulation. In the HEK 293-A R cells pre-treated with PKA inhibitor KT5720 and treated with NECA led to inhibit the IP and [Ca ] levels. The study distinctly demonstrated that A R modulates IP levels to release the [Ca ] via cAMP-dependent PKA. The role of A R mediated G pathway inducing IP mediated [Ca ] release may open new avenues in the therapy of neurodegenerative disorder.
ISSN:1872-8006