AbstractWithin species, levels of gene expression typically vary greatly between tissues, sexes, individuals, and populations. To investigate gene expression variation between sexes and populations in a single somatic tissue, we performed a quantitative analysis of the Malpighian tubule transcriptome in adult males and females of Drosophila melanogaster derived from two distinct populations (one from sub-Saharan Africa and one from northern Europe). We identified 2308 genes that differed in expression between the sexes and 2474 genes that differed in expression between populations at a false discovery rate of 5%. We also identified more than 1000 genes that showed a sex-by-population interaction in their expression. The genes that differed in expression between sexes showed enrichment for a wide variety of functions, although only 55% of them overlapped with sex-biased genes identified in whole-fly studies. The genes expressed differentially between populations included several that were previously implicated in adaptive regulatory evolution, an excess of cytochrome P450 genes, and many genes that were not detected in previous studies of whole flies. Our results demonstrate that there is abundant intraspecific gene expression variation within in a single, somatic tissue and uncover new candidates for adaptive regulatory evolution between populations.