Knockdown of GRK5 induces mitochondrial hyperpolarization in porcine embryos

Ferroptosis and mitochondrial hyperpolarization, both linked to oxidative stress, critically compromise embryo development during in vitro culture. This study demonstrates that GRK5 inhibition induces ferroptosis and mitochondrial hyperpolarization, leading to impaired developmental capacity in porcine embryos.

G-protein-coupled receptor kinase 5 (GRK5) regulates cellular processes including proliferation, differentiation, and apoptosis. This study explored the effects of double-stranded RNA microinjection and inhibitor treatment (5 and 10 μM) to suppress GRK5 expression on porcine embryo development. GRK5 inhibition impaired blastocyst development and quality, elevated p53 expression, and induced ferroptosis by reducing solute carrier family 7 member 11 and glutathione peroxidase 4. It also promoted apoptosis by downregulating B-cell lymphoma extra-large and upregulating Bcl-2 associated X. Moreover, GRK5 inhibition enhanced glutaminolysis, leading to increased adenosine triphosphate levels, mitochondrial membrane potential, and superoxide accumulation. Collectively, GRK5 inhibition increased p53 expression, induced apoptosis and ferroptosis, and promoted glutaminolysis, leading to mitochondrial hyperpolarization and impaired developmental capacity of porcine embryos.