A display substrate includes a base substrate, a first storage electrode, a second storage electrode, a first insulating layer, a first spacing member and a second spacing member. The base substrate includes a first pixel region and a second pixel region that are defined by a plurality of gate lines and a plurality of source lines extended in a direction different from an extended direction of the gate lines. The first storage electrode is in the first pixel region. The second storage electrode is in the second pixel region. The first insulating layer covers the first storage electrode, and has a recessed portion on the second storage electrode. The first spacing member is on the first insulating layer corresponding to the first storage electrode. The second spacing member is on the recessed portion.