title: The actin nucleator Cobl is crucial for Purkinje cell development and works in close conjunction with the F-actin binding protein Abp1.
abstract: Cortical actin dynamics shapes cells. To generate actin filaments, cells rely on actin nucleators. Cobl is a novel, brain-enriched, WH2 domain-based actin nucleator, yet, its functions remained largely elusive. Here, we reveal that Cobl plays a crucial role in Purkinje cell development using gene gun transfections within intact murine cerebellar contexts. Cobl deficiency impaired proper dendritic arborization of Purkinje cells and led to low-complexity arbors. Branch point numbers and density and especially higher order branching were strongly affected. Our efforts to reveal how Cobl is physically and functionally integrated into the cortical actin cytoskeleton showed that all Cobl loss-of-function phenotypes were exactly mirrored by knockdown of the F-actin-binding protein Abp1. By subcellular fractionations, protein interaction analyses, subcellular reconstitutions of protein complexes, colocalization studies in cells and tissues, and by functional analyses in neuronal morphogenesis we demonstrate that both proteins associate and work with each other closely. Cobl-mediated dendritic branch induction in hippocampal neurons critically relied on Abp1. Our study highlights that the functions of Abp1 are distinct from those of the Cobl-binding protein syndapin I. The importance of Cobl/Abp1 complex formation and of Abp1-mediated F-actin association was highlighted by functional rescue experiments demonstrating that a Cobl mutant deficient for Abp1 binding and an Abp1 mutant supporting Cobl association but lacking the F-actin binding ability failed to rescue the respective loss-of-function phenotypes. Thus, F-actin-anchored Cobl/Abp1 complexes seem crucial for neuromorphogenesis processes, particularly for the postnatal arborization of Purkinje cells representing the source for all motor coordination in the cerebellar cortex.
authors.affiliation: Institute for Biochemistry I, Jena University Hospital-Friedrich Schiller University Jena, D-07743 Jena, Germany.
authors.forename: Natja
authors.initials: N
authors.lastname: Haag
authors.forename: Lukas
authors.initials: L
authors.lastname: Schwintzer
authors.forename: Rashmi
authors.initials: R
authors.lastname: Ahuja
authors.forename: Nicole
authors.initials: N
authors.lastname: Koch
authors.forename: Julia
authors.initials: J
authors.lastname: Grimm
authors.forename: Heike
authors.initials: H
authors.lastname: Heuer
authors.forename: Britta
authors.initials: B
authors.lastname: Qualmann
authors.forename: Michael M
authors.initials: MM
authors.lastname: Kessels
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002531:Cerebellum / Q000254:growth & development* / Q000378:metabolism; D003598:Cytoskeletal Proteins; D003712:Dendrites / Q000648:ultrastructure; D055785:Gene Knockdown Techniques / Q000379:methods; D006624:Hippocampus / Q000166:cytology; D051379:Mice; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D057054:Molecular Imaging / Q000379:methods; D009154:Mutation; D055495:Neurogenesis / Q000502:physiology*; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011689:Purkinje Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D014162:Transfection / Q000379:methods; D018909:src Homology Domains / Q000235:genetics / Q000502:physiology*
chemical_list: D000199:Actins; C096541:Cobl protein, mouse; D003598:Cytoskeletal Proteins; C485512:Dbnl protein, mouse; D008840:Microfilament Proteins; D011506:Proteins