In situ potential mapping of space charge (SC) layer in a single GaN nanowire (NW) contacted to the Au metal electrode has been conducted using off-axis electron holography in order to study the space distribution of SC layer under electric biases. Based on the phase image reconstructed from the complex hologram the electrostatic potential at the SC layer was clearly revealed;the SC width was estimated to be about 76 nm under zero bias condition. In order to study dynamic interrelation between the SC layer and bias conditions, the variation of the electrostatic potential due to change of the SC widths respond to the different bias conditions have also been examined. The measured SC layers are found to vary between 68 nm and 91 nm, which correspond to the saturated SC layers at the GaN-Au contact under the forward and reverse bias conditions, respectively. By plotting the square widths of the SC layer against the applied voltages, donor density of GaN NWs was derived to be about 4.3*106 cm ? 3. Our experiments demon-strate that in-situ electron holography under electric field can be a useful method to investigate SC layers and donor density in single NW and other heterostructures.