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© The Rockefeller University Press, 0021-9525/2000//875 $5.00
The Journal of Cell Biology, Volume 149, Number 4, , 2000 875-888

Zebrafish vasa RNA but Not Its Protein Is a Component of the Germ Plasm and Segregates Asymmetrically before Germline Specification

^a Max Planck Institut für Entwicklungsbiologie Abteilung Genetik, 72076 Tübingen, Germany
^b Elektronenmikroskopie Labor, 72076 Tübingen, Germany
Max Planck Institut für Entwicklungsbiologie Abteilung Genetik, Spemannstrasse 35, 72076 Tübingen, Germany.49-7071-601-38449-7071-601-490

holger.knaut{at}tuebingen.mpg.de

Work in different organisms revealed that the vasa gene product is essential for germline specification. Here, we describe the asymmetric segregation of zebrafish vasa RNA, which distinguishes germ cell precursors from somatic cells in cleavage stage embryos. At the late blastula (sphere) stage, vasa mRNA segregation changes from asymmetric to symmetric, a process that precedes primordial germ cell proliferation and perinuclear localization of Vasa protein. Analysis of hybrid fish between Danio rerio and Danio feegradei demonstrates that zygotic vasa transcription is initiated shortly after the loss of unequal vasa mRNA segregation. Blocking DNA replication indicates that the change in vasa RNA segregation is dependent on a maternal program. Asymmetric segregation is impaired in embryos mutant for the maternal effect gene nebel. Furthermore, ultrastructural analysis of vasa RNA particles reveals that vasa RNA, but not Vasa protein, localizes to a subcellular structure that resembles nuage, a germ plasm organelle. The structure is initially associated with the actin cortex, and subsequent aggregation is inhibited by actin depolymerization. Later, the structure is found in close proximity of microtubules. We previously showed that its translocation to the distal furrows is microtubule dependent. We propose that vasa RNA but not Vasa protein is a component of the zebrafish germ plasm. Triggered by maternal signals, the pattern of germ plasm segregation changes, which results in the expression of primordial germ cell–specific genes such as vasa and, consequently, in germline fate commitment.

Key Words: primordial germ cells • Danio rerio • nebel • asymmetric segregation • RNA localization

© 2000 The Rockefeller University Press

Abbreviations used in this paper: dpf, days post fertilization; FMA, furrow microtubule array; hpf, hours postfertilization; MBT, midblastula transition; PGC, primordial germ cell; RACE, rapid amplification of cDNA ends; RNAPII, RNA polymerase II; UTR, untranslated region.

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