Depopulation and the emptying of settlements are a global process, which is largely represented in our country as well. Demographic changes in the structure of the population are more pronounced and intense in the settlements, which are far from the city centers and are located at higher altitudes. Depopulation, which is strongly present in the area of Southern Serbia, has the effect that some settlements are almost empty or have an older population and a very small share of young people. The aim of this paper is to present the population dynamics of the settlements of the City of Prokuplje in relation to their altitude in the period 1991-2022. For this reason, research was conducted based on the distribution of settlements in the City of Prokuplje into three groups according to altitude. The findings indicate a noticeable difference in the population dynamics of three groups of settlements and show to what extent the negative indicators of the number of inhabitants in the settlements are related to the altitude based on comparative and correlation analysis. The results provide insight into the state of demographic development and could help to create a more complete demographic picture of the City of Prokuplje.