DNA Microsatellites are codominant genetic markers and they are widely used in the characterization of biodiversity. Genetic variability of sarplaninian shepherds populations was determinate on the basis of analyses of ten DNA microsatellites loci (FH2361, DGN10, FH3287, FH3924, FH3608, FH3023, FH3489, FH3721, FH4027 and FH2141). In this study we have included 37 representative individuals from sarplaninian shepherd (19 lowland sarplaninians and 18 mountain sarplaninians). Informativnes of DNA microsatellites loci was determinate with parameter (Polymorphism Informative Content- PIC), and according to the determinate number of alleles for each locus. All ten DNA microsatellite loci were highly polymorphic. In the genome of sarplaninian shepherd loci FH2141 showed highest PIC value (0.975), and the loci with lowest PIC value was FH3924 locus (0.812). Number of detected alleles for each locus in the genome of sarplaninian shepherd varies from 16 (FH2361) to 33 (FH3023 and FH3489). Intrapopulation genetic variability was determinate according to the number alleles for each locus separately in each population, mean number of alleles for all eight loci and with number of the characteristic alleles. The most common alleles in populations of sarplaninian shepherds were detected in FH2141 loci, 49 (35 heterozygous and 14 homozygous). Taking in consideration the research results from this study a conclusion can be drawn that the genetic variability in Sarplaninian shepherd population is very low. This conclusion and similar stand points in the studies from a significant number of other authors, implicates that the Sarplaninian shepherd is relatively pure breed rarely interbred with phonotypic similar dogs in order to improve the characteristics of the breed.