The kinetics of dissolution of a two-dimensional chemisorbed layer made of uridine molecules adsorbed onto gold single-crystal electrodes have been studied by the potential step technique. Current transients have been recorded after single potential steps aimed at triggering the dissolution of the uridine chemisorbed monolayer formed on Au(111), Au(13,13,12) and Au(554). Irrespective of the electrode surface all the transients have satisfactorily been described by a model that combines a desorption step with a simultaneous hole nucleation and growth process. On the basis of a model considering an exponential law of nucleation and a surface diffusion-controlled growth, it is found that an increase of the step density of the electrode surface by affecting the ordering of the organic layer enhances markedly the rate of formation of holes in the film but slows down the expansion of the holes. The influence of the temperature and of the surfactant concentration on the nucleation and growth parameters is also reported.