In vertebrates, there are two variants of eukaryotic peptide elongation factor 1A (eEF1A; formerly eEF-$1{alpha}$), eEF1A1 and eEF1A2, which have three well-conserved domains ($D_I$, $D_{II}$, and $D_{III}$). In neurons, eEF1A1 is the embryonic type, which is expressed during embryonic development as well as the first two postnatal weeks. In the present study, EGFP-tagged eEF1A1 truncates were expressed in cortical neurons isolated from rat embryo (E18-19). Live cell images of transfected neurons showed that $D_{III}$-containing EGFP-fusion proteins (EGFP-$D_{III}$, -$D_{II-III}$, -$D_{I-III}$) formed clusters that were confined within somatodendritic domains, while $D_{III}$-missing ones (EGFP-$D_I$, -$D_{II}$, -$D_{I-II}$) and control EGFP were homogeneously dispersed throughout the neuron including axons. In dendrites, EGFP-$D_{III}$ was targeted to the heads of spine- and filopodia-like protrusions, where it was colocalized with $SynGAP{alpha}$, a postsynaptic marker. Our data indicate that $D_{III}$ of eEF1A1 mediates formation of clusters and localization to spines.