Identification of a 195 Kda protein in the striated rootlet: Its expression in ciliated and ciliogenic cells

Hagiwara, Haruo; Aoki, Takeo; Ohwada, Nobuo; Fujimoto, Toyoshi

doi:10.1002/(sici)1097-0169(200003)45:3<200::aid-cm3>3.0.co;2-4

Cited by 9 publications

(9 citation statements)

References 28 publications

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“…A similar localization of Dvl has been reported for Xenopus larvae (Park et al, 2008). Double immunostaining with R4109, a ciliary rootlet marker (Hagiwara et al, 2000), indicated that the Dvl2 was localized to the ciliary rootlets (Fig. 4F).…”

Section: Pcp Signals Regulate the Rotational But Not The Translationsupporting

confidence: 82%

“…The following antibodies were used: rabbit anti-pericentrin (Pcnt); (1:500; Covance, PRB-432C); goat anti-g-tubulin (1:100; Santa Cruz, sc7396); mouse CTR453 (1:10) (Bailly et al, 1989); mouse anti-b-catenin (1:200; BD 610154); chick anti-GFP (1:500; Aves, 2-P004-07D); rat anti-GFP (1:500; Nacalai, GF090R); rabbit anti-GFP (1:100; MBL); rabbit anti-Dvl2 (1:300; Santa Cruz, sc-13974); mouse anti-acetylated tubulin (1:1000; Sigma, T6793); rabbit anti-Myh9 (1:300; Covance, PRB-440P); rabbit anti-Myh10 (1:300; Covance, PRB-445P); rabbit anti-Myh14 (1:300; Covance, PRB-444P); rabbit pMLC2 (Myl9) (1:200; Cell Signaling, 3674); rat R4109 (1:2) (Hagiwara et al, 2000); mouse anti-ZO1 (1:300; Zymed, 33-9100); and species-specific secondary antibodies (Invitrogen or Jackson ImmunoResearch). Confocal images were obtained using LSM5-Pascal and LSM710 (Zeiss) microscopes.…”

Section: Histological Analysismentioning

confidence: 99%

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Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II

et al. 2010

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SUMMARYMotile cilia generate constant fluid flow over epithelial tissue, and thereby influence diverse physiological processes. Such functions of ciliated cells depend on the planar polarity of the cilia and on their basal bodies being oriented in the downstream direction of fluid flow. Recently, another type of basal body planar polarity, characterized by the anterior localization of the basal bodies in individual cells, was reported in the multiciliated ependymal cells that line the surface of brain ventricles. However, little is known about the cellular and molecular mechanisms by which this polarity is established. Here, we report in mice that basal bodies move in the apical cell membrane during differentiation to accumulate in the anterior region of ependymal cells. The planar cell polarity signaling pathway influences basal body orientation, but not their anterior migration, in the neonatal brain. Moreover, we show by pharmacological and genetic studies that non-muscle myosin II is a key regulator of this distribution of basal bodies. This study demonstrates that the orientation and distribution of basal bodies occur by distinct mechanisms.

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Section: Pcp Signals Regulate the Rotational But Not The Translationsupporting

confidence: 82%

Section: Histological Analysismentioning

confidence: 99%

Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II

et al. 2010

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“…They were preincubated with 2% gelatin in PBS for 15 min at 37°C, incubated with an anti-Golgi 58K antibody for 2 h at room temperature, washed, immersed in a 6 nm colloidal gold-conjugated anti-mouse IgG antibody for 2 h at room temperature, and processed for immunoelectron microscopy as in a previous report (Hagiwara et al 2000a).…”

Section: Indirect Immunofluorescence Microscopymentioning

confidence: 99%

Localization of Golgi 58K protein (formiminotransferase cyclodeaminase) to the centrosome

Hagiwara

Tajika

Matsuzaki

et al. 2006

Histochem Cell Biol

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In vertebrate cells, the centrosome consists of a pair of centrioles and surrounding pericentriolar material. Using anti-Golgi 58K protein antibodies that recognize formiminotransferase cyclodeaminase (FTCD), we investigated its localization to the centrosome in various cultured cells and human oviductal secretory cells by immunohistochemistry. In addition to the Golgi apparatus, FTCD was localized to the centrosome, more abundantly around the mother centriole. The centrosome localization of FTCD continued throughout the cell cycle and was not disrupted after Golgi fragmentation, which was induced by colcemid and brefeldin A. Centriole microtubules are polyglutamylated and stable against tubulin depolymerizing drugs. FTCD in the centrosome may be associated with polyglutamylated residues of centriole microtubules and may play a role in providing centrioles with glutamate produced by cyclodeaminase domains of FTCD.

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“…Cells were fixed in methanol for 5 min at -20°C or fixed with 1% paraformaldehyde in PHEM buffer (60 mM PIPES, 25 mM HEPES, 10 mM EGTA, 2 mM MgCl 2 , pH 6.9; Schliwa and van Blerkom 1981) for 5 min and extracted with 0.1% Triton X-100 for 90 s. Immunofluorescence staining was carried out as described previously (Hagiwara et al 2000b) using a rat monoclonal antibody R4109 against striated rootlets (Hagiwara et al 2000a), mouse monoclonal antibodies against α-tubulin, acetylated α-tubulin, and γ-tubulin (Sigma, St. Louis, Mo., USA), and fluorescein-isothiocyanate-conjugated goat anti-rat IgM (Immunotech, Marseille, France) or rhodamine-conjugated goat anti-mouse IgG (Jackson ImmunoResearch, West Grove, Pa., USA). TO-PRO-3 iodide (Molecular Probes, Eugene, Or., USA) was used for staining nuclei.…”

Section: Indirect Immunofluorescence Microscopymentioning

confidence: 99%

“…We have revealed the developmental course of striated rootlets during ciliogenesis (Hagiwara et al 1997), identified several proteins in the rootlets of human ciliated cells (Hagiwara et al 2000a), and analyzed their localization in the rootlets by immunohistochemistry (Hagiwara et al 2000b). However, little is known about the mechanism that regulates the organization of striated rootlets.…”

Section: Introductionmentioning

confidence: 99%

Depolymerization of microtubules by colcemid induces the formation of elongated and centriole-nonassociated striated rootlets in PtK 2 cells

Hagiwara

Takata

2002

Cell and Tissue Research

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Striated rootlets are cross-banded structures associated with the basal body, which extends the cilium. To determine whether microtubule dynamics influence the shape and distribution of striated rootlets, we have depolymerized the microtubules by colcemid and observed the rootlets by the immunohistochemical technique with the R4109 antibody that specifically reacts with a 195-kDa protein in the rootlets in PtK(2) cells. In control interphase cells, striated rootlets were observed in various profiles such as fibrillar, branched, or looped shapes and were associated with a pair of centrioles. Treatment with colcemid (0.1 micro g/ml or more) resulted in the elongation and/or structural complication of the centriole-associated rootlets and the organization of intracytoplasmic free rootlets. These changes appeared 6 h after colcemid treatment and became more prominent with time. The changes were reversible and almost disappeared 2 h after removal of the drug. Immunoelectron microscopy confirmed that the R4109 antibody decorated both centriole-associated rootlets and free rootlets. These findings indicate functional relationships between cytoplasmic microtubules and striated rootlets and the existence of rootlet-nucleating factors in the cytoplasm, in addition to centrioles.

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Identification of a 195 Kda protein in the striated rootlet: Its expression in ciliated and ciliogenic cells

Cited by 9 publications

References 28 publications

Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II

Planar polarity of multiciliated ependymal cells involves the anterior migration of basal bodies regulated by non-muscle myosin II

Localization of Golgi 58K protein (formiminotransferase cyclodeaminase) to the centrosome

Depolymerization of microtubules by colcemid induces the formation of elongated and centriole-nonassociated striated rootlets in PtK 2 cells

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