A display substrate includes an active pattern, a gate electrode, a first insulation layer and a pixel electrode. The active pattern is disposed on a base substrate. The active pattern includes a metal oxide semiconductor. The gate electrode overlaps the active pattern. The first insulation layer covers the gate electrode and the active pattern, and a contact hole is defined in the first insulation layer. The pixel electrode is electrically connected to the active pattern via the contact hole penetrating the first insulation layer. A first angle defined by a bottom surface of the first insulation layer and a sidewall of the first insulation layer exposed by the contact hole is between about 30° and about 50°.