The extracellular calcium‐sensing receptor promotes porcine egg activation via calcium/calmodulin‐dependent protein kinase II

Abstract Extracellular calcium is required for intracellular Ca 2+ oscillations needed for egg activation, but the regulatory mechanism is still poorly understood. The present study was designed to demonstrate the function of calcium‐sensing receptor (CASR), which could recognize extracellular calcium as first messenger, during porcine egg activation. CASR expression was markedly upregulated following egg activation. Functionally, the addition of CASR agonist NPS R‐568 significantly enhanced pronuclear formation rate, while supplementation of CASR antagonist NPS2390 compromised egg activation. There was no change in NPS R‐568 group compared with control group when the egg activation was performed without extracellular calcium addition. The addition of NPS2390 precluded the activation‐dependent [Ca 2+ ] i rise. When egg activation was conducted in intracellular Ca 2+ chelator BAPTA‐AM and NPS R‐568 containing medium, CASR function was abolished. Meanwhile, CASR activation increased the level of the [Ca 2+ ] i effector p‐CAMKII, and the presence of KN‐93, an inhibitor of CAMKII, significantly reduced the CASR‐mediated increasement of pronuclear formation rate. Furthermore, the increase of CASR expression following activation was reversed by inhibiting CAMKII activity, supporting a positive feedback loop between CAMKII and CASR. Altogether, these findings provide a new pathway of egg activation about CASR, as the extracellular Ca 2+ effector, promotes egg activation via its downstream effector and upstream regulator CAMKII.