title: Ttll9-/- mice sperm flagella show shortening of doublet 7, reduction of doublet 5 polyglutamylation and a stall in beating.
abstract: Nine outer doublet microtubules in axonemes of flagella and cilia are heterogeneous in structure and biochemical properties. In mammalian sperm flagella, one of the factors to generate the heterogeneity is tubulin polyglutamylation, although the importance of the heterogeneous modification is unclear. Here, we show that a tubulin polyglutamylase Ttll9 deficiency (Ttll9(-/-)) causes a unique set of phenotypes related to doublet heterogeneity. Ttll9(-/-) sperm axonemes had frequent loss of a doublet and reduced polyglutamylation. Intriguingly, the doublet loss selectively occurred at the distal region of doublet 7, and reduced polyglutamylation was observed preferentially on doublet 5. Ttll9(-/-) spermatozoa showed aberrant flagellar beating, characterized by frequent stalls after anti-hook bending. This abnormal motility could be attributed to the reduction of polyglutamylation on doublet 5, which probably occurred at a position involved in the switching of bending. These results indicate that mammalian Ttll9 plays essential roles in maintaining the normal structure and beating pattern of sperm flagella by establishing normal heterogeneous polyglutamylation patterns.
authors.affiliation: Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan.
authors.forename: Alu
authors.initials: A
authors.lastname: Konno
authors.affiliation: Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan International Mass Imaging Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Ikegami
authors.affiliation: Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Konishi
authors.affiliation: Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan.
authors.forename: Hyun-Jeong
authors.initials: HJ
authors.lastname: Yang
authors.affiliation: Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Niigata 9518585, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Abe
authors.affiliation: Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Niigata 9518585, Japan.
authors.forename: Maya
authors.initials: M
authors.lastname: Yamazaki
authors.affiliation: Department of Cellular Neurobiology, Brain Research Institute, Niigata University, Niigata 9518585, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Sakimura
authors.affiliation: Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan International Mass Imaging Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan.
authors.forename: Ikuko
authors.initials: I
authors.lastname: Yao
authors.affiliation: Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka 4150025, Japan.
authors.forename: Kogiku
authors.initials: K
authors.lastname: Shiba
authors.affiliation: Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka 4150025, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Inaba
authors.affiliation: Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan International Mass Imaging Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 4313192, Japan Department of Anatomy, The University of Hong Kong, 6/F, William MW Mong Block, 21 Sassoon Road, Pokfulam, Hong Kong SAR, China Division of Neural Systematics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, Aichi 4440867, Japan Riken Center for Molecular Imaging Science, Kobe, Hyogo 6500047, Japan setou@hama-med.ac.jp.
authors.forename: Mitsutoshi
authors.identifier: 0000-0002-1302-6467
authors.initials: M
authors.lastname: Setou
mesh_terms: D000818:Animals; D054468:Axoneme / Q000378:metabolism / Q000648:ultrastructure; D002452:Cell Count; D005260:Female; D005971:Glutamates / Q000378:metabolism*; D007248:Infertility, Male / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D010453:Peptide Synthases / Q000172:deficiency* / Q000378:metabolism; D013081:Sperm Motility / Q000502:physiology*; D013082:Sperm Tail / Q000502:physiology* / Q000648:ultrastructure; D013094:Spermatozoa / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D005971:Glutamates; D010453:Peptide Synthases; C112698:tubulin polyglutamylase
keywords: Axoneme; Flagella; Polyglutamylation; Sperm; Tubulin