hnRNP I is required to generate the Ca2+ signal that causes egg activation in zebrafish

Abstract

Egg activation is an important cellular event required to prevent polyspermy and initiate development of the zygote. Egg activation in all animals examined is elicited by a rise in free Ca2+ in the egg cytosol at fertilization. This Ca2+ rise is crucial for all subsequent egg activation steps, such as cortical granule exocytosis, which modifies the vitelline membrane to prevent polyspermy. The cytosolic Ca2+ rise is primarily initiated by inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ release from the endoplasmic reticulum. The genes involved in regulating the IP3-mediated Ca2+ release during egg activation remain largely unknown. Here we report on a zebrafish maternal-effect mutant, brom bones, which is defective in the cytosolic Ca2+ rise and subsequent egg activation events, including cortical granule exocytosis and cytoplasmic segregation. We show that the egg activation defects in brom bones can be rescued by providing Ca2+ or the Ca2+-release messenger IP3, suggesting that brom bones is a regulator of IP3-mediated Ca2+ release at fertilization. Interestingly, brom bones mutant embryos also display defects in dorsoventral axis formation accompanied by a disorganized cortical microtubule network, which is known to be crucial for dorsal axis formation. We provide evidence that the impaired microtubule organization is associated with non-exocytosed cortical granules from the earlier egg activation defect. Positional cloning of the brom bones gene reveals that a premature stop codon in the gene encoding hnRNP I (referred to here as hnrnp I) underlies the abnormalities. Our studies therefore reveal an important new role of hnrnp I in regulating the fundamental process of IP3-mediated Ca2+ release at egg activation.