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Fig. 1. EFHC1b is associated with various types of cilia. A: An RT-PCR analysis during early development (stages indicated) revealed that EFHC1b RNA levels increase at the onset of gastrulation, confirming previous data from Yanai et al. accessed through the Xenbase website. EF1α RNA was used as an internal control. In situ hybridization reveals the presence of EFHC1b RNA in the ciliated cells of the embryonic epidermis in stage 18 embryos (B) and in a number of different tissues in tailbud stage (stage 33) embryos (C)(CNS: central nervous system; NC: neural crest). D–F: RNAs encoding EFHC1b-GFP and Cetn2-RFP were injected into fertilized eggs and ectodermal explants were generated and analyzed when intact embryos reached stage 24; EFHC1b-GFP (D) was localized to the axonemes and excluded from the basal body regions of the cilia in multiciliated cells (E, F)(see Fig. 2). Scale bar of main image: 20 µm; scale bar for inserts: 10 µm. G–I: The gastrocoele roof plate regions of EFHC1b-GFP RNA injected, stage 19 embryos were dissected and probed with antibodies against GFP (G) and acetylated-α-tubulin (AAT)(H,I-merged image). Individual GRP cilia are indicated by arrowheads. Scale bar of main image: 15 µm; scale bar for inserts: 8 µm. J–L: At stage 26, a section through the neural tube region of an EFHC1b-GFP RNA injected embryos were stained for EFHC1b-GFP (J) and AAT (K, L-merged image); arrowheads indicate primary cilia. Other AAT structures may be neuronal axons or radial glia. Scale bar of main image: 10 µm; scale bar for inserts: 5 µm. |
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efhc1 (EF-hand domain (C-terminal) containing 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 32, lateral view, anterior right, dorsal up. |
