<i>Axin2</i> transcripts in chicken embryos from stage HH 23 to HH 32.

<p>(A) HH 23: expression pattern resembles what is described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0163610#pone.0163610.g002" target="_blank">Fig 2</a>. Limbs, inner ear, brain, eye, and <i>tb</i> express <i>Axin2</i> (A). (B, B.1 & B.2) HH 24: <i>Axin2</i> expression in the <i>ov</i> (B.1, white arrow) and <i>ba</i> (B.1), as well as in <i>nt</i> and <i>isf</i>(B.2, dorsal view). (C, C.1) HH 25: <i>Axin2</i> is expressed at similar embryonic structures with little change. (D, D.1, D.2, D.3) HH 26: <i>Axin2</i> expression in <i>ba</i> (D, black and red arrow; D.2), <i>ov</i>(D.2), <i>lb</i>(D.1), brain (D), eye (D), <i>isf</i> (D.3, white arrow) and <i>nt</i> (D.3). (E) HH27: <i>Axin2</i> transcripts in the facial whilst (E, white arrow), in the <i>ba</i> (black arrows), as well as in <i>lb</i>. The dorsal view (E.1) expression in the <i>nt</i> (white arrow). <i>Ba</i> display specific staining for c<i>Axin2</i> (HH 28: F & HH 29: G, white arrows). HH 28 and 29: intense expression in the embryonic shoulder (F & G, red arrows). Expression of <i>Axin2</i> in the forming interdigital spaces (F & G, black arrows). (H, H.1) HH 31: expression of <i>Axin2</i> mRNA in <i>lb</i> and apoptotic interdigital zones (H, black arrow), at the <i>ee</i>(H, red arrow) and in the <i>fb</i>(H & H.1, white arrows). (I, I.1) HH 32: <i>Axin2</i> transcripts in the eye (I, white arrow), <i>ee</i> (I, red arrow) and in the <i>fb</i> (I.1, white arrow). lb-limb buds, ov-otic vesicle, ba-branchial arches, tb-tail bud, nt-neural tube, isf-intersomitic furrow, fb-feather buds, ee-external ear.</p

Expression pattern of c<i>Axin2</i> from HH stage 17 to 22.

<p>(A) HH 17 embryo with <i>Axin2</i> expression in the brain, <i>lb</i> (red arrows), tail and <i>ov</i> (black arrow). (A.1) dorsal view: expression in the <i>dml</i> (white arrow). (B) HH 18: more prominent staining in the <i>lb</i> and in the <i>ba</i>. (C, C.1, C.2 & C.3) HH 19: <i>Axin2</i> transcripts in the <i>nt</i>(C.1) and <i>dml</i> (C.1, white arrow), in <i>lb</i> (C, C.1), in <i>ov</i> (C.2, white arrow), <i>ba</i> (C.2, black arrow) and in the tip of the tail (C.3, black arrow). (D, D.1, D.2) HH 20: similar expression of c<i>Axin2</i>. (D.1) the wing bud expresses <i>Axin2</i>, white arrow: <i>aer</i>. (D.2) dorsal view: prominent expression in <i>nt</i> (black arrowhead) and <i>isf</i>(white arrow). (E, E.1, E.2, E.3) HH 21: Transcripts in the tip of the tail (E, black arrowhead), in <i>nt</i> and <i>so</i>(E.1), in the otic anlage (E.2, black arrowhead) and in the developing <i>lb</i> (E.3). (F, F.1, F.2, F.3 and F.4) HH 22: consecutive expression of <i>Axin2</i> mRNA in <i>ba</i> (F, black arrow; F.3), <i>nt</i> (F.1, black arrow), <i>isf</i> (F, white arrow, F.2, white arrow) and tail (F.4, red arrow). lb-limb buds, ov-otic vesicle, dml-dorso medial lip, ba-branchial arches, nt-neural tube, aer-apical ectodermal ridge, isf-intersomitic furrow, so-somites.</p

Expression of <i>Axin2</i> in nephric duct.

<p><i>Axin2</i> mRNA expression in the <i>md</i> (HH 19: A:1, leg level, black arrow; HH 20: B.1, interlimb level, black arrow and B.2, leg level; HH23: C.1, caudal interlimb level, black arrow). The overlying thickened ectoderm strongly expresses <i>Axin2</i> from HH stage 20 (B.1 & B.2, black arrows). Pictures D.1 and E.1 demonstrate detectable transcripts in the cloacal ectoderm (HH 26: D.1, HH 28: E.1 black arrows). md-mesonephric duct.</p

<i>Axin2</i> expression during somitogenesis and in the developing neural tube.

<p>Transversal sections: green bars in overviews indicate sectioning level. (A) HH 15, overview; (A.1) expression in medial epithelial <i>so</i>(black arrow) and faintly all over the <i>nt</i>. (B) HH 16, overview; (B.1) increased expression in the medial somitic epithelium. (C) HH 17, overview; (C.1) <i>Axin2</i> in <i>dml</i> (black arrow), in the mesenchyme ventrally flanking the <i>nt</i> (red arrow) and predominantly in the dorsal <i>nt</i>. (C.2) Expression in dorso-medial somitic epithelium (black arrow), ventrally in the mesenchyme (red arrow) and all over the <i>nt</i>. (C.3) Expression in the medial epithelial <i>so</i>. (D) HH 19, overview; (D.1) expression in <i>dml</i> and <i>nt</i>; (D.2) expression restricted to dorsal <i>nt</i> (green arrow) and ventrally in the neighbouring tissue (red arrow). (E) HH 20, overview; (E.1) expression throughout the <i>dm</i>; (E.2) <i>Axin2</i> transcripts in the dorsal <i>nt</i> (green arrow), in the ventro-medially adjacent mesenchyme (red arrow) and weakly in the <i>dml</i>. (F) HH 22, overview; (F.1) <i>Axin2</i> in the dorsal-most <i>nt</i> (green arrow) and in the overlying ectoderm (black arrow). (G) HH27, overview; (G.1) <i>Axin2</i> expression in <i>rp</i>, <i>fp</i> (red arrow), <i>drg</i> (black arrow) and subectodermal space (green arrow). (H.1, H.2, H.3) <i>snt</i> of the tail. (H.1) HH 24, faint expression in all parts of the <i>snt</i>; (H.2) HH 27, restricted and intensified expression in the dorsal <i>snt</i> and in overlying subectodermal space; (H.3) HH 28, <i>Axin2</i> in dorsal most <i>snt</i> and subectodermal space. so-somite, nt-neural tube, dml-dorso medial lip, dm-dermomyotome, rp-roof plate, fp-floor plate, drg-dorsal root ganglion, snt-secondary neural tube.</p

<i>Axin2</i> transcripts in optic development.

<p>Transversal sections through the developing eye in <i>in-situ</i> hybridized embryos. (A.1) HH 15: the primary <i>ov</i> expresses <i>Axin2</i> in the proximal layer (black arrow). (B.1) HH 16: <i>Axin2</i> expression in both layers of the bi-layered secondary <i>oc</i>. (C.1) HH 17: <i>Axin2</i> expression in the proximal epithelium of the <i>oc</i> (black arrow). (D.1) HH 18: <i>Axin2</i> expression in the proximal layer of the <i>oc</i> (red arrow). The ectoderm surrounding the invaginating, <i>lv</i> (black arrow) expresses <i>Axin2</i>. (D.2) HH 18: A section through the rostral most part of the <i>oc</i>, with highest expression rate in these margins. (E.1) HH 19: the proximal layer of the <i>lv</i> expresses <i>Axin2</i> (red arrow) as well as the <i>oc</i> in both layers adjacent to the vesicular space (black arrow). (F.1, F.2, F.3, F.4) HH 20: expression in the ectoderm surrounding the eye, in the directly underlying mesenchyme, in the epithelium of the <i>oc</i> towards the lumen (F.1, black arrow; F.4, black arrow) and in the lens epithelium (F.1, F.2, F.3, red arrows). (HH 23: G.1; HH 24: H.1 and H.2, black arrows; HH 25: I.1; HH 26: J.1, J.2, J.3; HH 27: K.2, black arrow, HH 28: L.1, L.2, black arrows) <i>Axin2</i> expression in the ectoderm and subectodermal mesenchyme (future cornea). (HH 25: I.1, black arrow; HH 26: J.1, black arrow, HH 27: K.1, black arrow) c<i>Axin2</i> expression in the developing <i>on</i> and <i>och</i>. (H.3, HH 24; K.2, HH 27) expression of <i>Axin2</i> in the developing lens (red and green arrows). ov-optic vesicle, oc-optic cup, lv-lens vesicle, on-optic nerve, och-optic chiasm, rpe-retinal pigmented epithelium.</p

Expression of <i>Axin2</i> in chicken limb development.

<p>(A.1, wing bud, w) HH 17, strong expression of <i>Axin2</i> in the limb, predominantly in the dorsal most part (black arrow). (A.2) The leg bud (l) shows less staining, a patch in the ventro-proximal tissue expresses <i>Axin2</i> (black arrow). (B.1, leg bud) HH 18, transcripts in the dorsal mesenchyme and in the distal ectoderm. (C.1) HH 19, expression in the dorsal and most proximal mesenchyme of the wing bud. (HH 19, C.1; HH 20, D.1 black arrow; D.2) <i>Axin2</i> expression <i>aer</i>. (C.1) HH 19 and (D.1) HH 20, <i>Axin2</i> in dorsal subectodermal mesenchyme, in ectoderm and in the <i>aer</i>. (E.1, E.2) HH 23; (F.1) HH 25: Transcripts in the ectoderm and <i>aer</i> of the limb buds. (E.2–I.3) HH 23 to HH 28: the ectoderm expresses high levels of <i>Axin2</i>. (G.1) HH 26, black arrow; (I.3) HH 28, black arrow: the margins of the developing <i>bo</i> show transcripts of <i>Axin2</i>. aer-apical ectodermal ridge, bo-bones.</p

Expression of c<i>Axin2</i> in stages HH 11 to HH 15.

<p>Overviews (A, B, C) and transversal sections of chicken embryos. Green bars in overviews indicate sectioning level (C). (A) HH 11: <i>Axin2</i> expression in the head, <i>pnp</i>, <i>nt</i> and (A, A.1) <i>dml</i> of the <i>so</i> (black arrow). (B) HH 14: strong expression in the <i>dml</i> (black arrow). (B.1) transversal section through the head at mid-brain level with transcripts in the medial head fold in in the adjacent neighbouring mesenchyme (black arrow). (C) HH 15: <i>Axin2</i> expression in the brain (white arrow), the <i>dml</i> (C.1 black arrow) and in the <i>pnp</i>. (C.2) expression in the <i>snc</i> and in the ventral <i>snt</i>. (C.3, C.4) <i>Axin2</i> expression in the <i>tbm</i>. (C.5) section through head and neck with expression of <i>Axin2</i> in the <i>oc</i>, the neighbouring <i>hb</i> and the <i>ov</i>. pnp-posterior neuropore, nt-neural tube, dml-dorso-medial lip, so-somites, snc-secondary notochord, snt-secondary neural tube, tbm-tail bud mesoderm, oc-otic cup, hb-hind-brain, ov-optic vesicle.</p

Additional file 1 of Multiple transcriptome analyses reveal mouse testis developmental dynamics

Supplementary Material

Additional file 1: of Inflammation and neuronal death in the motor cortex of the wobbler mouse, an ALS animal model

Comparison between Iba-1-labeled microglial cells in the motor cortex of WT mice and intense symptomatic WR mice 60 d.p.n. (A–C) Relation of activated microglial cells (red) and neurons labeled with neuronal nuclei antibody (green) in brain tissue of WT mice. (D–F) Relation of activated microglial cells (red) and neurons labeled with neuronal nuclei antibody (green) in the brain tissue of severely symptomatic WR mice. (Scale bar = 20 μm). (TIF 10223 kb

Additional file 2: of Inflammation and neuronal death in the motor cortex of the wobbler mouse, an ALS animal model

Visualization of the weak staining of Iba-1- and TNF-α-labeled microglial and caspase 3-positive neuronal cells in motor cortex tissue of WR mice 20 d.p.n. (A–C) Relation of activated microglial cells (red) and neurons labeled with neuronal nuclei antibody (green) in brain tissue of non-symptomatic WR mice. (D–F) Iba-1-labeled microglial cells (red) synthesizing the cytokine TNF-α (green). (G–H) Caspase 3-positive (red) neurons labeled with NeuN (neuronal nuclei antibody) (green). (Scale bar = 20 μm). (TIF 15107 kb