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Fig.â1. Whole mount in situ hybridization showing spatial and temporal expression of tweety homologs in developing X. laevis embryos. Row 1 and 2: dorsal views. Rows 3, 4, 5: both lateral (top embryo, with dorsal side on top) and dorsal (bottom embryo) views. Anterior is oriented to the right for all embryos. Row 6: close up of anterior part of the embryo, with a lateral view (dorsal side on top) and a dorsal view (with anterior directed upwards). (A, G, M) Stages 19â20 (late neurula). (B, H, N) Stages 22â23 (early tailbud). (C, I, O) Stages 25â26 (mid-tailbud). (D, J, P) Stages 29â31 (late tailbud). (E, K, Q) Stages 34â35 (swimming tadpole). (F, L, R) Stages 34â35 (swimming tadpole, close up). (AâF) Ttyh1 expression. (GâL) Ttyh2 expression. (MâR) Ttyh3 expression. Arrowheads indicate regions of expression. (fb, forebrain; e, eye; hb, hindbrain; mb, midbrain; ot, otic vesicle; pa, pharyngeal arches; sc, spinal cord; nt, neural tube; so, somites; Roman numerals: cranial ganglia.) Scale barsâ=â1.0âmm.
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Fig. 2. Histological analysis of tweety homolog expression in X. laevis stages 35â37 (swimming tadpole) using whole-mount in situ hybridization. Transverse 18 μm sec- tions along the anteriorâposterior axis with dorsal to the top. (AâO) 10à magnification images focused on neural regions with signal. (Aâ²âOâ²) 4à magnification images to show to the absence of signal in other regions. (A, F, K) Sections showing forebrain. (B, G, L) Sections showing midbrain. (C, H, M) Sections showing hindbrain. (D, I, N) Sections showing anterior spinal cord. (E, J, O) Sections showing posterior spinal cord. (AâE) Ttyh1 expression. (FâJ) Ttyh2 expression. (KâO) Ttyh3 expression. Arrowheads indicate regions of expression. (asc, anterior spinal cord; mb, midbrain; hb, hindbrain; fb, forebrain; e, eye; nc, neural crest; ot, otic vesicle; psc, posterior spinal cord; sc, spinal cord; Roman numerals: cranial ganglia.) Scale bars = 0.25 mm.
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Fig. 3. qRT-PCR showing ttyh gene expression from single cell to stage 45. Stage 0 in the figure denotes unfertilized eggs. At each stage gene expression was quantitated using the δδCT method and corrected for expression of the control gene (Drosha). Relative expression is displayed log10 transformed.
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Fig.â4. Due to the concentrated expression of ttyh2 in the cranial ganglia, double fluorescent in situ hybridization was carried out to determine its co-localization with another known cranial ganglia marker. Histological analysis of tweety homolog 2 (ttyh2) (Cy3-tyramide) and xVGlut1 (FITC-tyramide) expression in X. laevis stage 35 (swimming tadpole) using whole-mount fluorescent in situ hybridization. Transverse 25âµm sections along the anteriorâposterior axis with dorsal to the top. (A) Section showing forebrain. (B) Section showing midbrain. (C) Section showing hindbrain. (mb, midbrain; hb, hindbrain; fb, forebrain; e, eye; Roman numerals: cranial ganglia.) Scale barsâ=â0.25âmm.
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ttyh1 (tweety family member 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 34-35, lateral view, anterior right, dorsal up, (left) , and dorsal view (right).
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ttyh1 (tweety family member 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25-26, lateral view, anterior right, dorsal up (above), and dorsal view (below).
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ttyh1 (tweety family member 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19-20, dorsal view, anterior right.
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ttyh3 (tweety family member 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25-26, dorsal view, anterior right.
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ttyh3 (tweety family member 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19-20, dorsal view, anterior right.
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