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. 2014 Dec 19;8:147. doi: 10.3389/fncir.2014.00147

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Copyright © 2014 Hsieh, Ulrich, Issa, Wan and Papazian.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Expression of Nav1.6 and Kv3.3 coincides with the emergence of spontaneous tonic firing in Purkinje cells. (A) Whole mount double fluorescent in situ hybridization was performed on TLN zebrafish at 3, 4, 5, and 6 dpf using digoxigenin-labeled aldoca (Zebrin-II, red) and fluorescein-labeled scn8aa (Nav1.6, green) antisense probes (Thisse and Thisse, 2008; Brend and Holley, 2009). Projected confocal image stacks for aldoca, scn8aa, and merged images are shown in the 3 left-most panels, respectively. Dashed white boxes in merged images denote regions of one cerebellar hemisphere that have been enlarged and shown in the right-most panel (zoom). Here and in subsequent parts of the figure, dorsal views are shown. Anterior is to the left. Scale bars: 40 μm. (B) Enlarged single optical sections from the projections in Figure 2A at 4, 5, and 6 dpf are shown. Arrowheads identify individual Purkinje cell bodies. Scale bars: 10 μm. (C) Whole mount double fluorescent in situ hybridization was performed on TLN zebrafish at 5 dpf using digoxigenin-labeled aldoca (Zebrin-II, red, left panel) and fluorescein-labeled scn8aa (Nav1.6, green, center panel) sense probes (Thisse and Thisse, 2008; Brend and Holley, 2009). Merged image is shown in right panel. Scale bar: 40 μm. (D) Whole mount double fluorescent in situ hybridization was performed on TLN zebrafish at 3, 4, 5, and 6 dpf using digoxigenin-labeled aldoca (Zebrin-II, red) and fluorescein-labeled kcnc3a (Kv3.3, green) antisense probes (Thisse and Thisse, 2008; Brend and Holley, 2009). Projected confocal image stacks for aldoca, kcnc3a, and merged images are shown in the 3 left-most panels, respectively. Dashed white boxes in merged images denote regions of one cerebellar hemisphere that have been enlarged and shown in the right-most panel (zoom). Scale bars: 40 μm. (E) Enlarged single optical sections from the projections in Figure 2D at 4, 5, and 6 dpf are shown. Arrowheads identify individual Purkinje cell bodies. Scale bars: 10 μm. (F) Whole mount double fluorescent in situ hybridization was performed on TLN zebrafish at 5 dpf using digoxigenin-labeled aldoca (Zebrin-II, red, left panel) and fluorescein-labeled kcnc3a (Kv3.3, green, center panel) sense probes (Thisse and Thisse, 2008; Brend and Holley, 2009). Merged image is shown in right panel. Scale bar: 40 μm.