Ashwin Gene Expression Profiles in Oocytes, Preimplantation Embryos, and Fetal and Adult Bovine Tissues

SIMPLE SUMMARY: Ashwin is a gene involved in the morphogenesis of the central nervous system and the early embryonic development of Xenopus laevis. The analysis of its phylogeny in silico has shown that its functions are restricted to vertebrates, but we lack additional information regarding its biological importance in higher vertebrates, such as mammals. The present study reveals the wide and variable expression of this gene in different bovine organs and confirms its significant expression during early embryonic development, with a pattern similar to that reported for maternal genes. In addition, specific expression of this gene throughout follicular development and during bovine spermatogenesis is revealed, leading to the proposal of ashwin as a new gene with important biological implications in bovine development and reproduction. ABSTRACT: The ashwin gene, originally identified in Xenopus laevis, was found to be expressed first in the neural plate and later in the embryonic brain, eyes, and spinal cord. Functional studies of ashwin suggest that it participates in cell survival and anteroposterior patterning. Furthermore, ashwin is expressed zygotically in this species, which suggests that it participates in embryonic development. Nevertheless, the expression of this gene has not been studied in mammals. Thus, the aim of this study was to analyze the ashwin expression pattern in bovine fetal and adult tissues, as well as in three independent samples of immature and mature oocytes, and in two- to four-, and eight-cell embryos, morula, and blastocysts. Spatiotemporal expression was analyzed using real-time polymerase chain reaction (PCR); ashwin mRNA was detected in all tissues analyzed, immature and mature oocytes, and two- to eight-cell embryos. It was down-regulated in morula and blastocysts, suggesting that this expression profile is similar to that of maternal genes. Immunohistochemical localization of the ashwin protein in fetal and adult ovaries and testes reveals that this protein is consistently present during all stages of follicular development and during bovine spermatogenesis. These observations lead us to propose ashwin as an important gene involved in mammalian reproduction.