A display substrate includes a base substrate, a switching element, a protecting layer, an organic layer, a first pixel electrode and a second pixel electrode. The switching element is on the base substrate, and includes a gate electrode, a source electrode and a drain electrode. The protecting layer is on the switching element, and includes a first hole exposing the drain electrode. The organic layer is on the protecting layer, and includes a second hole which exposes a side surface of the protecting layer which defines the first hole and exposes a top surface of the protecting layer which is adjacent to the side surface of the protecting layer. The first pixel electrode is on the organic layer. The second pixel electrode overlaps the first pixel electrode, and is electrically connected to the drain electrode via the first and second holes.