title: Maternal and zygotic expression of a nanos-class gene in the leech Helobdella robusta: primordial germ cells arise from segmental mesoderm.
abstract: The nanos-class gene of the leech Helobdella robusta (Hro-nos) is present as a maternal transcript whose levels decay during cleavage; HRO-NOS protein is more abundant in the D quadrant cells relative to the A, B, and C quadrants; and HRO-NOS is more abundant in the ectodermal precursor cell (DNOPQ) than in its sister mesodermal precursor (DM) (Pilon and Weisblat, 1997). Here, using in situ hybridization, we show that Hro-nos mRNA is broadly distributed throughout the zygote, is concentrated in both animal and vegetal teloplasm during stage 1 and is at higher levels in DNOPQ than in DM at stage 4b. Hro-nos expression increases after stage 7, as judged by in situ hybridization, developmental RT-PCR, and western blots; this increase must therefore represent later zygotic expression. Of particular interest, during stages 9 and 10, each of 11 mid-body segments (M8-M18) has a pair of Hro-nos positive "spots" comprising of one or two large cells each. These spots later disappear in an anteroposterior progression. We find that these Hro-nos-expressing cells are of mesodermal origin, arising in a segmentally iterated manner from the M lineage, and correspond to cells previously proposed as primordial germ cells (PGCs; Bürger, 1891; Weisblat and Shankland, 1985). These results support the proposal that nanos-class genes functioned in the specification of germline cells in the ancestral bilaterian and possibly in a separate process related to embryonic polarity in the ancestral protostome.
authors.affiliation: Department of Biological Sciences, Stanford University, Stanford, CA 94305-5020, USA.
authors.forename: Dongmin
authors.initials: D
authors.lastname: Kang
authors.forename: Marc
authors.initials: M
authors.lastname: Pilon
authors.forename: David A
authors.initials: DA
authors.lastname: Weisblat
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D029721:Drosophila Proteins; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology*; D017403:In Situ Hybridization; D019476:Insect Proteins / Q000235:genetics*; D007865:Leeches / Q000196:embryology / Q000235:genetics*; D008648:Mesoderm / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015053:Zygote / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D029721:Drosophila Proteins; D019476:Insect Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C070574:nos protein, Drosophila