Parkinson's disease (PD) is a neurodegenerative disorder characterized by the aggregation of -synuclein protein and selective death of dopaminergic (DA) neurons in the substantia nigra of the midbrain. Although the molecular pathogenesis of PD is not completely understood, a recent study has reported that eukaryotic translation elongation factor 1 alpha (eEF1A) declined in the PD-affected brain. Therefore, the roles of eEF1A1 and eEF1A2 in the prevention of DA neuronal cell death in PD are aimed to be investigated. Herein, by using <i>Caenorhabditis elegans</i> as a PD model, we investigated the role of <i>
eft-3/eft-4</i>, the worm homolog of <i>eEF1A1/eEF1A2</i>, on 6-hydroxydopamine (6-OHDA)-induced DA neuron degeneration. Our results demonstrated that the expressions of e<i>ft-3</i> and <i>
eft-4</i> were decreased in the 6-OHDA-induced worms. RNA interference (RNAi) of <i>
eft-3</i> and <i>
eft-4</i> resulted in dramatic exacerbation of DA neurodegeneration induced by 6-OHDA, as well as aggravated the food-sensing behavior, ethanol avoidance, and decreased lifespan when compared with only 6-OHDA-induced worms. Moreover, downregulation of <i>
eft-3/4</i> in 6-OHDA-induced worms suppressed the expression of the anti-apoptotic genes, including <i>PI3K/age-1, PDK-1/pdk-1, mTOR/let-363</i>, and <i>AKT-1,2/akt-1,2</i>, promoting the expression of apoptotic genes such as <i>BH3/egl-1</i> and <i>Caspase-9/ced-3.</i> Collectively, these findings indicate that eEF1A plays an important role in the 6-OHDA-induced neurodegeneration through the phosphatidylinositol 3-kinase (PI3K)/serine/threonine protein kinase (Akt)/mammalian target of rapamycin (mTOR) pathway and that eEF1A isoforms may be a novel and effective pro-survival factor in protective DA neurons against toxin-induced neuronal death.