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  • Slow calcium waves mediate furrow microtubule reorganization and germ plasm compaction in the early zebrafish embryo.

Slow calcium waves mediate furrow microtubule reorganization and germ plasm compaction in the early zebrafish embryo.

Development (Cambridge, England) (2018-04-11)
Celeste Eno, Timothy Gomez, Diane C Slusarski, Francisco Pelegri
ABSTRACT

Zebrafish germ plasm ribonucleoparticles (RNPs) become recruited to furrows of early zebrafish embryos through their association with astral microtubules ends. During the initiation of cytokinesis, microtubules are remodeled into a furrow microtubule array (FMA), which is thought to be analogous to the mammalian midbody involved in membrane abscission. During furrow maturation, RNPs and FMA tubules transition from their original distribution along the furrow to enrichments at the furrow distal ends, which facilitates germ plasm mass compaction. We show that nebel mutants exhibit reduced furrow-associated slow calcium waves (SCWs), caused at least in part by defective enrichment of calcium stores. RNP and FMA distal enrichment mirrors the medial-to-distal polarity of SCWs, and inhibition of calcium release or downstream mediators such as Calmodulin affects RNP and FMA distal enrichment. Blastomeres with reduced or lacking SCWs, such as early blastomeres in nebel mutants and wild-type blastomeres at later stages, exhibit medially bundling microtubules similar to midbodies in other cell types. Our data indicate that SCWs provide medial-to-distal directionality along the furrow to facilitate germ plasm RNP enrichment at the furrow ends.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Calmodulin Antibody, Upstate®, from mouse
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
D-myo-Inositol 1,4,5-tris-phosphate trisodium salt, ≥95% (by 1H NMR and TLC)