vasa and piwi are required for mitotic integrity in early embryogenesis in the spider Parasteatoda tepidariorum

• DOI: 10.1016/j.ydbio.2014.08.032
• Publication Date: 2014-09-23T09:07:43Z
• Authors: Evelyn E. Schwager; Yue Meng; Cassandra G. Exta...
• Keywords: 570; Embryo, Nonmammalian; Spindle; Achaearanea; Molecular Sequence Data; 590; Mitosis; Cell Cycle Proteins; vasa; Polymerase Chain Reaction; DEAD-box RNA Helicases; 03 medical and health sciences; piwi; RNA interference; Image Processing, Computer-Assisted; Animals; Chelicerata; Cloning, Molecular; Molecular Biology; In Situ Hybridization; Phylogeny; DNA Primers; 0303 health sciences; Base Sequence; Cell Biology; Sequence Analysis, DNA; Immunohistochemistry; Germ Cells; Argonaute Proteins; Oocytes; RNA Interference; Sequence Alignment; Developmental Biology

Studies in vertebrate and invertebrate model organisms on the molecular basis of primordial germ cell (PGC) specification have revealed that metazoans can specify their germ line either early in development by maternally transmitted cytoplasmic factors (inheritance), or later in development by signaling factors from neighboring tissues (induction). Regardless of the mode of PGC specification, once animal germ cells are specified, they invariably express a number of highly conserved genes. These include vasa and piwi, which can play essential roles in any or all of PGC specification, development, or gametogenesis. Although the arthropods are the most speciose animal phylum, to date there have been no functional studies of conserved germ line genes in species of the most basally branching arthropod clade, the chelicerates (which includes spiders, scorpions, and horseshoe crabs). Here we present the first such study by using molecular and functional tools to examine germ line development and the roles of vasa and piwi orthologues in the common house spider Parasteatoda (formerly Achaearanea) tepidariorum. We use transcript and protein expression patterns of Pt-vasa and Pt-piwi to show that primordial germ cells (PGCs) in the spider arise during late embryogenesis. Neither Pt-vasa nor Pt-piwi gene products are localized asymmetrically to any embryonic region before PGCs emerge as paired segmental clusters in opisthosomal segments 2-6 at late germ band stages. RNA interference studies reveal that both genes are required maternally for egg laying, mitotic progression in early embryos, and embryonic survival. Our results add to the growing body of evidence that vasa and piwi can play important roles in somatic development, and provide evidence for a previously hypothesized conserved role for vasa in cell cycle progression.