Halleran AD et al. (2015), Characterization of tweety gene (ttyh1-3) expre...

XB-ART-50234

Gene Expr Patterns 2015 Jan 01;171:38-44. doi: 10.1016/j.gep.2014.12.002.

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Characterization of tweety gene (ttyh1-3) expression in Xenopus laevis during embryonic development.

Halleran AD , Sehdev M , Rabe BA , Huyck RW , Williams CC , Saha MS .

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The tweety family of genes encodes large-conductance chloride channels and has been implicated in a wide array of cellular processes including cell division, cell adhesion, regulation of calcium activity, and tumorigenesis, particularly in neuronal cells. However, their expression patterns during early development remain largely unknown. Here, we describe the spatial and temporal patterning of ttyh1, ttyh2, and ttyh3 in Xenopus laevis during early embryonic development. Ttyh1 and ttyh3 are initially expressed at the late neurula stage are and primarily localized to the developing nervous system; however ttyh1 and ttyh3 both show transient expression in the somites. By swimming tadpole stages, all three genes are expressed in the brain, spinal cord, eye, and cranial ganglia. While ttyh1 is restricted to proliferative, ventricular zones, ttyh3 is primarily localized to postmitotic regions of the developing nervous system. Ttyh2, however, is strongly expressed in cranial ganglia V, VII, IX and X. The differing temporal and spatial expression patterns of ttyh1, ttyh2, and ttyh3 suggest that they may play distinct roles throughout embryonic development.

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Species referenced: Xenopus laevis
Genes referenced: slc17a7 ttyh1 ttyh2 ttyh3

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References [+] :

Al-Jumaily, Expression of three distinct families of calcium-activated chloride channel genes in the mouse dorsal root ganglion. 2007, Pubmed

	Fig. 1. 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 19–20 (late neurula). (B, H, N) Stages 22–23 (early tailbud). (C, I, O) Stages 25–26 (mid-tailbud). (D, J, P) Stages 29–31 (late tailbud). (E, K, Q) Stages 34–35 (swimming tadpole). (F, L, R) Stages 34–35 (swimming tadpole, close up). (A–F) Ttyh1 expression. (G–L) Ttyh2 expression. (M–R) 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 bars = 1.0 mm.
	Fig. 2. Histological analysis of tweety homolog expression in X. laevis stages 35–37 (swimming tadpole) using whole-mount in situ hybridization. Transverse 18 μm sec- tions along the anterior–posterior axis with dorsal to the top. (A–O) 10× magnification images focused on neural regions with signal. (A′–O′) 4× 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. (A–E) Ttyh1 expression. (F–J) Ttyh2 expression. (K–O) 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.
	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.
	Fig. 4. 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 25 µm sections along the anterior–posterior 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 bars = 0.25 mm.
	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).
	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).
	ttyh1 (tweety family member 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19-20, dorsal view, anterior right.




	ttyh3 (tweety family member 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25-26, dorsal view, anterior right.
	ttyh3 (tweety family member 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19-20, dorsal view, anterior right.