The first member of the RF-amide peptide superfamily to be characterized, in 1977, was the cardioexcitatory peptide, FMRFamide, isolated from the ganglia of the clamMacrocallista nimbosa (1). Since then, a large number of such peptides, designated after their C-terminal arginine (R) and amidated phenylalanine (F) residues, have been identified in representative species of all major phyla. The discovery, 12 years ago, that the RF-amide peptide kisspeptin, acting via GPR54, was essential for the onset of puberty and reproduction, has been a major breakthrough in reproductive physiology (2–4). It has also put in front of the spotlights RF-amide peptides and has invigorated research on this superfamily of regulatory neuropeptides. The present Research Topic aims at illustrating major advances achieved, through comparative studies in (mammalian and nonmammalian) vertebrates and invertebrates, in the knowledge of RF-amide peptides in terms of evolutionary history and physiological significance. Since 2006, by means of phylogenetic analyses, the superfamily of RFamide peptides has been divided into five families/groups in vertebrates (5, 6): kisspeptin, 26RFa/QRFP, GnIH (including LPXRFa and RFRP), NPFF, and PrRP. Recent data reveal that SIFamide-type neuropeptides in protostomian invertebrates and SALMFamide-type neuropeptides in deuterostomian invertebrates share a common evolutionary origin with vertebrate LPXRFa and PQRFa (7). Comparative studies on non-mammalian vertebrates and invertebrates allow major advances in the knowledge of the evolutionary history of the RF-amide peptide superfamily. Such phylogenetical studies also contribute to refine classification and nomenclature of both peptides and receptors. In this issue, Yun et al. (8) show that the concept of coevolution of peptide ligands and their cognate receptors helps to re-examine not only the classification of receptors but also their peptides. They thus report that kisspeptin should be classified in the galanin/spexin family rather than in the RF-amide peptide family. Another example is given by Tachibana and Sakamoto (9) who propose non-mammalian PrRP (C)-RFa to be renamed PrRP2. With the identification of the QRFPR genes in coelacanth and spotted gar, Larhammar et al. (10) demonstrate that the QRFP system is complex in the early stages of vertebrate evolution and secondarily becomes restricted in mammals. In their review, Elphick and Mirabeau (11) recount the occurrence of the RFamide motif in bilaterian neuropeptide families. They report that peptides, such as NPY/NPF, have acquired modified C-terminal characteristics in vertebrates, while RFamide-type peptides like luqins have been lost in the vertebrate lineage. They also underline some neuropeptide families (e.g., CCK/sulfakinins) in which the RFamide motif is unique to protostomian members. Osugi et al. (12) show that identification of GnIH in agnathans (lamprey) and amphioxus reveals that the C-terminal amide motif of GnIH can differ, being QPQRF or RPQRF, in addition to previously observed LPXRF in birds, mammals and most of fish, and MPQRF in grass puffer and medaka. As indicated above, the characterization of the first RF-amide peptidewas carried out in amollusk. Since then, many different genes have been identified in invertebrates, and the reviews by ZatylnyGaudin and Favrel (13) in mollusks, and by Li and Kim (14) and Peymen et al. (15) in nematodes, emphasize the need of identifying receptors for these peptides in invertebrates and characterizing their signaling pathways.