A display substrate includes an insulating substrate, a signal line, first and second pixel electrodes, a connection line and an insulating layer. The signal line is disposed on the insulating substrate. The first pixel electrode is electrically connected to the signal line through a switching element. The second pixel electrode overlaps the first pixel electrode. The connection line contacts an end portion of the signal line and extends to an end portion of the insulating substrate. The insulating layer is disposed between the first and second pixel electrodes and covers the connection line. The connection line is protected by the insulating layer, and the reliability of the display substrate is enhanced.