title: Identification and expression pattern analysis of Piwi genes during the spermiogenesis of Portunus trituberculatus.
abstract: The Piwi genes have an important role in stem cell development, gametogenesis and RNA interference in diverse organisms. So far, most of the studies have focused on the function of Piwis in vertebrates, but their function during spermiogenesis in invertebrates still remains largely unclear. In order to investigate the function of Piwis during spermiogenesis in the crab Portunus trituberculatus, we use RT-PCR and RACE to identify three Piwi complete cDNA sequences from the total RNA of the testis in P. trituberculatus. The deduced amino acid sequences of P. trituberculatus Piwi-1, Piwi-2 and Piwi-3 showed that each contains a well-conserved PAZ domain and PIWI domain. RT-PCR analyzed the tissue expression pattern of P. trituberculatus Piwi-1, Piwi-2 and Piwi-3 in the testis, heart, muscle, hepatopancreas and gill. All of the Piwis are found in germ cells of adult testis in P. trituberculatus by in situ hybridization, suggesting that these genes may play function during spermiogenesis in this species.
authors.affiliation: The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China; School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, China.
authors.forename: Dong-Fang
authors.initials: DF
authors.lastname: Xiang
authors.affiliation: School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, China.
authors.forename: Jun-Quan
authors.initials: JQ
authors.lastname: Zhu
authors.affiliation: The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Cong-Cong
authors.initials: CC
authors.lastname: Hou
authors.affiliation: The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China. Electronic address: wxyang@spermlab.org.
authors.forename: Wan-Xi
authors.initials: WX
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003386:Brachyura / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D017434:Protein Structure, Tertiary / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013091:Spermatogenesis / Q000235:genetics*
chemical_list: D018076:DNA, Complementary
keywords: 3-Aminopropyl-triethoxysilane; 5-bromo-4-chloro-3-indolylphosphate; A; AC; APES; AT; Ago3; Argonaute3; BCIP; DEPC; DIG; Digoxigenin; EV; GSP; IL; MC; ML; NBT; NJ; NUP; Nested Universal Primer; OL; ORF; P-element induced wimpy testis; PAZ; PBS; PFA; PG; PM; PV; Piwi; Piwi–Argonaute–Zwille; Portunus trituberculatus; RA; RACE; RT-PCR; SSC; Spermiogenesis; TD-PCR; Testis; UPM; UTR; Universal Primer Mix; aa; acrosomal tubule; acrosome; amino acids; apical cap; diethylpyrocarbonate; endoplasmic reticulum vesicle; gene specific primer; inner layer; membrane complex; middle layer; neighbor-joining; nitroblue tetrazolium chloride; open reading frame; outer layer; paraformaldehyde; phosphate-buffered saline; piRNA; piwi-interacting RNA; plasma membrane; proacrosomal granule; proacrosomal vesicle; radial arms; rapid-amplification of cDNA ends; reverse transcription-polymerase chain reaction; standard saline citrate; touchdown PCR; untranslated region