Rsph9 is critical for ciliary radial spoke assembly and central pair microtubule stability

Zhu, Lei; Líu, Hao; Chen, Yawen; Yan, Xiumin; Zhu, Xueliang

doi:10.1111/boc.201800060

Cited by 21 publications

(23 citation statements)

References 37 publications

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“…Our video microscopy results showed that the movement of beating cilia bundles is characterized by lower amplitude and disorderly rotation beating pattern instead of orderly planar beating pattern in subventricular zone en-face of Rsph9 −/− mice. Liu et al reported that knockdown of RSPH9 by RNAi in mouse ependymal cells, which differentiated from radial glia ex vivo, resulted in a near complete CP loss 26 ; however, our results showed that there were no defects with CP in Rsph9 −/− ependymal motile cilia in vivo, and few exhibited various ultrastructure disorganizations. This may be due to the truncated RSPH9, which may still support the central pair formation, or the differences between in vivo and in vitro experiments.…”

Section: Discussioncontrasting

confidence: 81%

Loss of Rsph9 causes neonatal hydrocephalus with abnormal development of motile cilia in mice

Zou

Liu

et al. 2020

Sci Rep

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Hydrocephalus is a brain disorder triggered by cerebrospinal fluid accumulation in brain cavities. Even though cerebrospinal fluid flow is known to be driven by the orchestrated beating of the bundled motile cilia of ependymal cells, little is known about the mechanism of ciliary motility. RSPH9 is increasingly becoming recognized as a vital component of radial spokes in ciliary "9 + 2" ultrastructure organization. Here, we show that deletion of the Rsph9 gene leads to the development of hydrocephalus in the early postnatal period. However, the neurodevelopment and astrocyte development are normal in embryonic Rsph9 −/− mice. The tubular structure of the central aqueduct was comparable in Rsph9 −/− mice. Using high-speed video microscopy, we visualized lower beating amplitude and irregular rotation beating pattern of cilia bundles in Rsph9 −/− mice compared with that of wild-type mice. And the centriolar patch size was significantly increased in Rsph9 −/− cells. TEM results showed that deletion of Rsph9 causes little impact in ciliary axonemal organization but the Rsph9 −/− cilia frequently had abnormal ectopic ciliary membrane inclusions. In addition, hydrocephalus in Rsph9 −/− mice results in the development of astrogliosis, microgliosis and cerebrovascular abnormalities. Eventually, the ependymal cells sloughed off of the lateral wall. Our results collectively suggested that RSPH9 is essential for ciliary structure and motility of mouse ependymal cilia, and its deletion causes the pathogenesis of hydrocephalus. Hydrocephalus is a prevalent birth defect triggered by excessive accumulation of cerebrospinal fluid (CSF) in brain cavities 1. Natural CSF flow is secreted by specialized ependymal cells in the choroid plexuses of lateral ventricles. In addition, the CSF carries signaling molecules, nutrients, microRNAs and exosomes 2-4. It passes through the 3rd ventricles, cerebral aqueduct, and the 4th ventricles and can be absorbed by arachnoid granulations 5,6. The directional flow of CSF is driven by continuous CSF secretion and by the orchestrated beating of bundles of motile cilia that are located at the apical surface of ependymal cells 7. The motile cilia are arranged structurally as a "9 + 2" axoneme, which is composed of nine interconnecting peripheral pairs of microtubules and a central pair of single microtubules. The radial spoke head protein, RSPH9, is a structural protein located at the T-shaped macromolecular configurations protruding from the nine peripheral pairs of microtubules 8. Interaction between radial spoke heads and the central pair of single microtubules is the central for ciliary regulation. Previous studies showed that Chlamydomonas RSP9 mutant strains lack the entire radial spoke head complex and displacement of the central pair of single microtubules 9. And zebrafish Rsph9 mutant larvae exhibit similar cilia-dysmotility defects and reduced initiation of the acoustic startle response 10. However, mutations in RSPH9 cause primary ciliary dyskinesia (PCD; MIM 244400) in human, which is cha...

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Section: Discussioncontrasting

confidence: 81%

Loss of Rsph9 causes neonatal hydrocephalus with abnormal development of motile cilia in mice

Zou

Liu

et al. 2020

Sci Rep

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“…Unless otherwise 393 indicated, the culture medium was Dulbecco's Modified Eagle's medium (DMEM) 394 supplemented with 10% fetal bovine serum (Biochrom, Cambridge, UK), 0.3 mg/ml 395 glutamine (Sigma), 100 U/ml penicillin (Invitrogen), and 100 U/ml streptomycin 396 (Invitrogen). Multiciliated mouse ependymal cells were obtained and cultured as 397 described (Zhu et al, 2019). Briefly, the telencephalon taken from the P0 C57BL/6J 398 mice were digested and spread onto laminin coated flasks.…”

Section: Cell Culture and Immunofluorescent 392mentioning

confidence: 99%

“…We first examined whether all the mouse homologous RS head proteins (Rsph1, 4a, 6a, 96 9 and 10b) and RSP3 (Rsph3b) localize in the axoneme of motile cilia in multiciliated 97 mouse ependymal cells (mEPCs) (Delgehyr et al, 2015;Zhu et al, 2019). We found 98 that all these subunits except Rsph6a can be localized in the cilia of mEPCs ( Figure 99 1A).…”

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confidence: 99%

Distinct architecture and composition of mouse axonemal radial spoke head revealed by cryo-EM

Zheng

Ding

et al. 2019

Preprint

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1The radial spoke (RS) transmits mechanochemical signals from the central pair 2 apparatus (CP) to axonemal dynein arms to coordinate ciliary motility. The RS head, 3 directly contacting with CP, differs dramatically in morphology between protozoan and 4 mammal. Here we show the murine RS head is compositionally distinct from the 5 Chlamydomonas one. Our reconstituted murine RS head core complex consists of 6 Rsph1, Rsph3b, Rsph4a, and Rsph9, lacking Rsph6a whose orthologue exists in the 7Chlamydomonas RS head. We present the unprecedented cryo-EM structure of RS head 8 core complex at 4.5-Å resolution and identified the subunit location and their 9 interaction network. In this complex, Rsph3b, Rsph4a, and Rsph9 forms a compact 10 body with Rsph4a serving possibly as an assembly scaffold and Rsph3b in a location 11 that might link the head with stalk. Interestingly, two Rsph1 subunits constitute the two 12 stretching-arms possibly for optimized RS-CP interaction. We also propose a sawtooth 13 model for the RS-CP interaction. Our study suggests that the RS head experiences 14 profound remodeling to probably comply with both structural and functional alterations 15 of the axoneme during evolution. 16

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“…We observed that the ciliary localizations of multiple CP proteins, including CP MTs-associated Spef1, C2 MT-associated Hydin, C1 MT-associated Spag6, and C1-C2 bridge-associated Spag16 (Fig. 2a, S3) 22,35,44 , were abolished or markedly reduced in Wdr47 -/-mEPCs, whereas Rsph4a, a radial spoke component 45,46 , was not affected ( Fig. 2b; Fig.…”

Section: Wdr47 Is Critical For Both Ciliary Localization Of Cp Proteimentioning

confidence: 92%

Wdr47, Camsaps, and Katanin cooperate to generate ciliary central microtubules

Líu

Zheng

Zhu

et al. 2020

Preprint

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The axonemal central pair (CP) are non-centrosomal microtubules critical for planar ciliary beat. How they form, however, is poorly understood. Here, we show that mammalian CP formation requires cooperative activities of Katanin, Camsaps, and Wdr47. Katanin severs peripheral microtubules to produce central microtubule seeds in nascent cilia. Camsaps stabilize minus ends of the seeds to facilitate MT outgrowth, whereas Wdr47 concentrates Camsaps into the axonemal central lumen to properly position the central microtubules. Wdr47 deficiency in mouse multicilia results in complete loss of CP, rotatory beat, and primary ciliary dyskinesia. Overexpression of Camsaps induces central microtubules in Wdr47-/- ependymal cells but at the expense of low efficiency, abnormal numbers, and wrong location, whereas overexpression of a dominant inhibitor of Katanin impairs the CP formation. We propose that Wdr47, Camsaps, and Katanin constitute a general cooperative work team for the generation of non-centrosomal MT arrays in polarized subcellular compartments.

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Rsph9 is critical for ciliary radial spoke assembly and central pair microtubule stability

Cited by 21 publications

References 37 publications

Loss of Rsph9 causes neonatal hydrocephalus with abnormal development of motile cilia in mice

Loss of Rsph9 causes neonatal hydrocephalus with abnormal development of motile cilia in mice

Distinct architecture and composition of mouse axonemal radial spoke head revealed by cryo-EM

Wdr47, Camsaps, and Katanin cooperate to generate ciliary central microtubules

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