title: Endocytic recycling protein EHD1 regulates primary cilia morphogenesis and SHH signaling during neural tube development.
abstract: Members of the four-member C-terminal EPS15-Homology Domain-containing (EHD) protein family play crucial roles in endocytic recycling of cell surface receptors from endosomes to the plasma membrane. In this study, we show that Ehd1 gene knockout in mice on a predominantly B6 background is embryonic lethal. Ehd1-null embryos die at mid-gestation with a failure to complete key developmental processes including neural tube closure, axial turning and patterning of the neural tube. We found that Ehd1-null embryos display short and stubby cilia on the developing neuroepithelium at embryonic day 9.5 (E9.5). Loss of EHD1 also deregulates the ciliary SHH signaling with Ehd1-null embryos displaying features indicative of increased SHH signaling, including a significant downregulation in the formation of the GLI3 repressor and increase in the ventral neuronal markers specified by SHH. Using Ehd1-null MEFS we found that EHD1 protein co-localizes with the SHH receptor Smoothened in the primary cilia upon ligand stimulation. Under the same conditions, EHD1 was shown to co-traffic with Smoothened into the developing primary cilia and we identify EHD1 as a direct binding partner of Smoothened. Overall, our studies identify the endocytic recycling regulator EHD1 as a novel regulator of the primary cilium-associated trafficking of Smoothened and Hedgehog signaling.
authors.affiliation: The Department of Pathology &Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.|Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Sohinee
authors.initials: S
authors.lastname: Bhattacharyya
authors.affiliation: Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Rainey
authors.affiliation: The Department of Genetics, Cell Biology &Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.|Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Arya
authors.forename: Bhopal C
authors.lastname: Mohapatra
authors.forename: Insha
authors.lastname: Mushtaq
authors.affiliation: The Department of Biochemistry &Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Samikshan
authors.initials: S
authors.lastname: Dutta
authors.affiliation: Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Manju
authors.initials: M
authors.lastname: George
authors.affiliation: Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Storck
authors.affiliation: The Department of Pathology &Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Rodney D
authors.initials: RD
authors.lastname: McComb
authors.affiliation: The Department of Pathology &Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: David
authors.initials: D
authors.lastname: Muirhead
authors.affiliation: The Department of Genetics, Cell Biology &Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Gordon L
authors.initials: GL
authors.lastname: Todd
authors.affiliation: The Department of Genetics, Cell Biology &Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Gould
authors.affiliation: The Department of Biochemistry &Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Kaustubh
authors.initials: K
authors.lastname: Datta
authors.affiliation: Department of Pharmacology, Creighton University School of Medicine, Omaha, NE, USA.
authors.forename: Janee
authors.initials: J
authors.lastname: Gelineau-van Waes
authors.affiliation: The Department of Genetics, Cell Biology &Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.|Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.|Fred and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Vimla
authors.initials: V
authors.lastname: Band
authors.affiliation: The Department of Pathology &Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.|The Department of Genetics, Cell Biology &Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.|Eppley Institute for Research in Cancer and Allied Diseases,University of Nebraska Medical Center, Omaha, NE, USA.|Fred and Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA.
authors.forename: Hamid
authors.initials: H
authors.lastname: Band
mesh_terms: D000818:Animals; D002923:Cilia / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D047108:Embryonic Development / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D005804:Genes, Lethal; D000068617:Genetic Background; D005838:Genotype; D053823:Hedgehog Proteins / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009024:Morphogenesis / Q000235:genetics; D005810:Multigene Family; D054259:Neural Tube / Q000196:embryology* / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D015398:Signal Transduction; D000072086:Smoothened Receptor / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000235:genetics*
chemical_list: C485640:Ehd1 protein, mouse; D053823:Hedgehog Proteins; C491022:Smo protein, mouse; D000072086:Smoothened Receptor; D033921:Vesicular Transport Proteins