The traditional morphologicaily grounded placement of South American guinea pig-like rodents (Caviomorpha) within one of the two rodent suborders, Hystricognathi, has been disputed by recent analysis of protein and nucleic acid sequence data. The Caviomorpha and possibly all Hystricognathi would be considered a separate order, distinct from the other rodent suborder, Sciurognathi, and thus of the order Rodentia, and would be placed closer phylogenetically to other mammals [Graur, D., Hide, W. A. & Li, W.-H. (1991) Nature (London) 351,[649][650][651][652]. To address the discrepancy between morphological comparisons and sequence analyses, we have applied an alternative form of molecular analysis. We demonstrate that BC1 RNA, a neural-specific small cytoplasmic RNA that is the product of a retropositionally generated gene (a gene derived by reverse transcription of RNA followed by insertion of the DNA copy into the genome), is present in Sciurognathi and guinea pig but not in other mammalian orders including Lagomorpha, Artiodactyla, and Primates.The species:confined, tissue-specific expression of a retroposed sequence therefore supports the morphological evidence for monophyly of Rodentia inclusive of guinea pig and demonstrates the usefulness of such molecular genetic markers. Furthermore, the conservation and tissue-specific expression of the BC1 RNA gene in the two divergent rodent suborders suggests that this macromolecule has been exapted into a functional role (i.e., coopted into a variant or novel function) in the rodent nervous system. Based on a large body of morphological evidence, the order Rodentia, containing approximately half of all mammalian species (1), has been subdivided into two suborders (2). The Sciurognathi include true rats, mice, hamsters, and squirrellike rodents, while the Hystricognathi, also representing a group of Old and New World rodent families, are now primarily restricted to Africa and South America. Examples of this diverse suborder include guinea pig, chinchilla, and porcupine. However, challenging this morphologically established phylogenetic scheme of Rodentia is an apparent "molecular paradox"-namely, most guinea pig protein and nucleic acid sequences exhibit an unusually high level of dissimilarity when compared with their rodent orthologues (refs. 3-5, but also see ref. 6).A recently proposed solution (7, 8) would define Caviomorpha and possibly the entire suborder Hystricognathi as a separate mammalian order distinct from Rodentia. Unfortunately, the proposal of a separate order addresses the molecular data but leads to incongruence with paleontological information and anatomical data (9). Thus, well-established morphological synapomorphies between Sciurognathi and Hystricognathi would be reinterpreted as homoplasies (10). Furthermore, the molecular evidence underlying this exThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to ...