A display substrate includes a gate electrode on a base substrate, an active pattern which overlaps the gate electrode and includes a metal oxide semiconductor, an insulation pattern on the active pattern, a source electrode which contacts the active pattern, a drain electrode which contacts the active pattern and is spaced apart from the source electrode, and a first passivation layer which covers the active pattern and the insulation pattern, and includes fluorine, where the active pattern includes a first portion which directly contacts the insulation pattern and overlaps the gate electrode and the insulation pattern, a second portion which contacts the first passivation layer and has an electrical conductivity substantially larger than that of the first portion, a third portion which contacts the first passivation layer, has an electrical conductivity substantially larger than that of the first portion and is spaced apart from the second portion.