Figure 6.
Kif6 deficiency impairs the planar polarity of multicilia across the ependyma. Quantification results were pooled together (mean ± SD plus sample dots) and subjected to unpaired two-tailed Student’s t test. Mi, Mild hydrocephalus; Se, Severe hydrocephalus. (A) Experimental scheme. Whole-mounts of ventricle walls were freshly dissected and soaked in a culture medium. The ex-vivo tissues were stained with SiR-tubulin, a fluorescent dye for MTs, to label cilia. A fixed z-plane was imaged at 10-ms intervals to visualize ciliary motilities. (B–E) Motilities of multicilia in ependymal tissues of the indicated genotypes. Representative image sequences (B and D) show ciliary motilities in two consecutive beat cycles. Color-coded arrows indicate directions of effective and recovery strokes, respectively. Color images are overlaid pseudo-colored image sequences covering the first beat cycle. Multiciliary beat frequencies were quantified from five pairs of littermates (60 cells per mouse) (C) or three pairs of littermates (50 cells per mouse) (E). No gross differences were observed in the back-and-forth beat pattern of multicilia (B and D) and their beat frequencies (C and E) between Kif6+/+ and Kif6−/− or Kif9+/+ and Kif9−/− littermates. (F)Kif6 deficiency abolished the planar polarity of ependymal multicilia. The first three consecutive frames cropped from Video 9 were pseudocolored and overlaid to show ciliary motilities. Beat directions of multicilia are indicated by arrows. (G)Kif9 deficiency did not impair the planar polarity of ependymal multicilia. (H) Schematic for assays on multicilia-driven liquid flows. Fresh ependymal tissues were positioned on glass-bottomed culture dishes with the multicilia-containing side down. Fluorescent beads were added into culture medium. Beads in a fixed z-plane beneath ex-vivo ependymal tissues were imaged at 100-ms intervals. (I) Liquid flows and flow directions are shown by merging image sequences in the first 500 ms of Video 10. Colored arrows indicate trajectories of representative, rapidly moving fluorescent puncta during the period of time. (J) Quantification results for bead velocities. Data were from two pairs of P36 and one pair of P48 littermates (60 traceable beads for each mouse).

Kif6 deficiency impairs the planar polarity of multicilia across the ependyma. Quantification results were pooled together (mean ± SD plus sample dots) and subjected to unpaired two-tailed Student’s t test. Mi, Mild hydrocephalus; Se, Severe hydrocephalus. (A) Experimental scheme. Whole-mounts of ventricle walls were freshly dissected and soaked in a culture medium. The ex-vivo tissues were stained with SiR-tubulin, a fluorescent dye for MTs, to label cilia. A fixed z-plane was imaged at 10-ms intervals to visualize ciliary motilities. (B–E) Motilities of multicilia in ependymal tissues of the indicated genotypes. Representative image sequences (B and D) show ciliary motilities in two consecutive beat cycles. Color-coded arrows indicate directions of effective and recovery strokes, respectively. Color images are overlaid pseudo-colored image sequences covering the first beat cycle. Multiciliary beat frequencies were quantified from five pairs of littermates (60 cells per mouse) (C) or three pairs of littermates (50 cells per mouse) (E). No gross differences were observed in the back-and-forth beat pattern of multicilia (B and D) and their beat frequencies (C and E) between Kif6+/+ and Kif6−/− or Kif9+/+ and Kif9−/− littermates. (F)Kif6 deficiency abolished the planar polarity of ependymal multicilia. The first three consecutive frames cropped from Video 9 were pseudocolored and overlaid to show ciliary motilities. Beat directions of multicilia are indicated by arrows. (G)Kif9 deficiency did not impair the planar polarity of ependymal multicilia. (H) Schematic for assays on multicilia-driven liquid flows. Fresh ependymal tissues were positioned on glass-bottomed culture dishes with the multicilia-containing side down. Fluorescent beads were added into culture medium. Beads in a fixed z-plane beneath ex-vivo ependymal tissues were imaged at 100-ms intervals. (I) Liquid flows and flow directions are shown by merging image sequences in the first 500 ms of Video 10. Colored arrows indicate trajectories of representative, rapidly moving fluorescent puncta during the period of time. (J) Quantification results for bead velocities. Data were from two pairs of P36 and one pair of P48 littermates (60 traceable beads for each mouse).

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