Figure 6

Normal ciliary beating and ependymal flow in Zfta^tm1/tm1 mice. (A) Schematic of the live imaging of ciliary beating. Freshly isolated wholemount preparations of the lateral walls of the LV were stained with FITC-labelled CD24, washed with DMEM culture medium and stapled to a dissection dish filled with DMEM. Ciliary beating was observed with a water immersion objective and a high-speed camera. (B) The CBF was unaffected in Zfta^tm1/tm1 mice at P31–35. Data shown are the mean ± SD. Forty-four cells from 4 control mice, 44 cells from 3 ZFTA^tm1/tm1 mice, p = 0.89. N.S., not significant. Each point on the graph is the CBF of an individual E1 cell. See also Movies S1–S2. (C) Schematic of the ependymal flow assay. A glass needle was filled with fluorescent beads (green) and placed onto the wholemount preparations of the lateral walls of the LV. A, anterior; D, dorsal; P, posterior; PA, point of adhesion; V, ventral. (D) The speeds of the fluorescent beads on the wholemount preparation are expressed as the mean ± SD. Each point on the graph represents the speed of individual fluorescent beads. Control; n = 25 beads, 5 mice. Zfta^tm1/tm1; n = 20 beads, 4 mice, p = 0.15. See also Movies S3–S4. (E–H) Coronal MRI scans showing the lateral (E, F) and fourth (G, H) ventricles in the control (E, G) and Zfta^tm1/tm1 (F, H) mice at P31–35. White arrows indicate the lateral (E, F) and fourth (G, H) ventricles. Bar = 5 mm. (I, J) Quantification of the ventricular volumes in LV (I) and 4 V (J). Data shown are the mean ± SD. Each point on the graph represents the volume of ventricles in individual mice. Control (cont.); n = 8 mice: Zfta^tm1/tm1; n = 8 mice; p = 0.28 (LV) and 0.64 (4 V). (K, K′, L, L′) Chain migration of new neurons is not affected in the Zfta^tm1/tm1 V-SVZ. Migrating new neurons were stained with anti-DCX antibody in the control (K, K′) and Zfta^tm1/tm1 (L, L′) wholemount preparations of the lateral walls of the LV at P31. The insets in K and L are magnified in K′ and L′, respectively. Bars = 1 mm. *Bubble in mounting media.