title: aura (mid1ip1l) regulates the cytoskeleton at the zebrafish egg-to-embryo transition.
abstract: Embryos from females homozygous for a recessive maternal-effect mutation in the gene aura exhibit defects including reduced cortical integrity, defective cortical granule (CG) release upon egg activation, failure to complete cytokinesis, and abnormal cell wound healing. We show that the cytokinesis defects are associated with aberrant cytoskeletal reorganization during furrow maturation, including abnormal F-actin enrichment and microtubule reorganization. Cortical F-actin prior to furrow formation fails to exhibit a normal transition into F-actin-rich arcs, and drug inhibition is consistent with aura function promoting F-actin polymerization and/or stabilization. In mutants, components of exocytic and endocytic vesicles, such as Vamp2, Clathrin and Dynamin, are sequestered in unreleased CGs, indicating a need for CG recycling in the normal redistribution of these factors. However, the exocytic targeting factor Rab11 is recruited to the furrow plane normally at the tip of bundling microtubules, suggesting an alternative anchoring mechanism independent of membrane recycling. A positional cloning approach indicates that the mutation in aura is associated with a truncation of Mid1 interacting protein 1 like (Mid1ip1l), previously identified as an interactor of the X-linked Opitz G/BBB syndrome gene product Mid1. A Cas9/CRISPR-induced mutant allele in mid1ip1l fails to complement the originally isolated aura maternal-effect mutation, confirming gene assignment. Mid1ip1l protein localizes to cortical F-actin aggregates, consistent with a direct role in cytoskeletal regulation. Our studies indicate that maternally provided aura (mid1ip1l) acts during the reorganization of the cytoskeleton at the egg-to-embryo transition and highlight the importance of cytoskeletal dynamics and membrane recycling during this developmental period.
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Room 2455 Genetics, Madison, WI 53706, USA.
authors.forename: Celeste
authors.initials: C
authors.lastname: Eno
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Room 2455 Genetics, Madison, WI 53706, USA.
authors.forename: Bharti
authors.initials: B
authors.lastname: Solanki
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Room 2455 Genetics, Madison, WI 53706, USA fjpelegri@wisc.edu.
authors.forename: Francisco
authors.initials: F
authors.lastname: Pelegri
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002966:Clathrin / Q000378:metabolism; D048749:Cytokinesis / Q000235:genetics*; D003594:Cytoplasmic Granules / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D034281:Dynamins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
chemical_list: D000199:Actins; D002966:Clathrin; D003598:Cytoskeletal Proteins; D034281:Dynamins; D050988:Vesicle-Associated Membrane Protein 2; D029961:Zebrafish Proteins; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C000622645:mid1ip1l protein, zebrafish
keywords: Cortical granules; Cytokinesis; Cytoskeleton; F-actin; Membrane exocytosis; Mid1; Mid1ip1l; Opitz G/BBB syndrome; Zebrafish