The G-protein-coupled receptors (GPCRs) are a large superfamily of seven transmembrane domain-spanning proteins that mediate signal transduction by activation of G-proteins. Mammalian GPCR genes are reputed to be largely intron-deficient, but there have been no reports of using genome-wide analyses of gene structure to investigate this. Using complete genome sequences, we analysed the intron content of over 850 members of the rhodopsin-like GPCR subfamily (family A G-protein-coupled receptor or GPCR-A) in four species. We find that mouse and human GPCR-As have a large and significant reduction in intron number compared to the rest of their genome. In contrast, invertebrate GPCR-As have an intron repertoire similar to, or slightly greater than, the rest of their genome, suggesting that the reduced intron content in mammals is due to widespread intron loss. Furthermore, we provide a specific example of intron loss through analysis of an intron that is conserved in position and phase within a phylogenetically diverse range of GPCR-As within six vertebrate and invertebrate species. Together, these two lines of evidence provide compelling evidence for the widespread loss of introns during the evolution of the mammalian GPCR-A family.