Apolipoprotein C1 (Apoc1) is associated with lipoprotein metabolism, but its physiological role during embryogenesis is largely unknown. We reveal a new function of Apoc1b, a transcript isoform of Apoc1, in epiboly during zebrafish gastrulation. Apoc1b is expressed in yolk syncytial layers and in deep cells of the ventral and lateral region of the embryos. It displays a radial gradient with high levels in the interior layer and low levels in the superficial layer. Knockdown of Apoc1b by injecting antisense morpholino (MO) caused the epiboly arrest in deep cells. Moreover, we show that the radial intercalation and the radial gradient distribution of E-cadherin are disrupted both in Apoc1b knockdown and overexpressed embryos. Therefore, Apoc1b controls epiboly via E-cadherin-mediated radial intercalation in a gradient-dependent manner.