In this study, V-shaped dislocations in a GaN epitaxial layer on a free-standing GaN substrate were observed. Our investigation further revealed that the V-shaped dislocations were newly generated at the interface in the epilayer rather than propagated from the GaN substrate. V-shaped dislocations consist of two straight parts. The straight parts of the V-shaped dislocations were separated from each other in the m-direction and tilted toward the step-flow direction of the GaN epitaxial layer. The V-shaped dislocations are continuous single dislocations having a Burgers vector component of 1a and an intrinsic stacking fault between their straight parts.