Many amphibian species possess male-specific secretory breeding glands. In anurans, such specialized glands are often present as nuptial pads at the first digit of the hand, which are pressed against the female during amplexus. However, the role of nuptial pad secretions remains largely unknown. Here, we investigate the inner morphology as well as the RNA expression patterns of the nuptial pads of several Central American Lithobates species (Ranidae). As shown for the breeding glands of other amphibian species, the Lithobates nuptial pads are composed of large specialized mucus glands, excreting proteinaceous content to the surface. Whole-transcriptome sequencing revealed that for one of the species the most highly expressed transcripts encoding secretory proteins in the nuptial pads are sodefrin precursor-like factor (SPF) proteins, known to have a pheromone function in multiple amphibian species. The other two species, however, lack high expression of SPF transcripts but express other secretory proteins, whose roles in amphibian breeding glands remain unknown. Several of these proteins are highly expressed in all samples, suggesting a direct role during courtship or for the maintenance/development of the glands.