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. 2013 Aug 30;4:112. doi: 10.3389/fendo.2013.00112

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Copyright © 2013 Wakayama, Katow and Katow.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or 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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Immunochemical expression pattern of Epith-2 during early development of H. pulcherrimus. (A) Immunoblotting pattern. Lane 1; unfertilized eggs (uf), lane 2; fertilized eggs (f), lane 3; the 2-cell stage embryos (2cs), lane 4; the 4-cell stage embryos (4cs), lane 5; the 16-cell stage embryos (16cs), lane 6; morulae (Mor), lane 7; swimming blastulae (sBl), lane 8; mesenchyme blastulae (mBl), lane 9; mid-gastrulae (mG), lane 10; late gastrulae (lG), lane 11; prism larvae (Prsm), lane 12; pluteus larvae (Plut). Epith-2 is detected consistently at the 143 kDa region from unfertilized eggs to prism larvae, but a new band is detected at the 126 kDa region at the pluteus stage, which replaced the larger band of relative molecular mass. (B) Whole-mount double-stained immunohistochemical expression pattern of Epith-2 (green) and DNA with propidium iodide (red) (a–j’) and Polywax sagittal 6-μm thick sections (g”). (a) A 4-μm thick optical section of an unfertilized egg (uf). (a’) Higher magnification image of the box in (a). Arrow, anti-Epith-2 mAb-positive egg surface. (b) A 4-μm thick optical section of a fertilized egg (f). (b’) Higher magnification image of the box in (b). (c) A 4-μm thick optical section of a two-cell embryo (2cs). (c’) Higher magnification image of the box in (c). (d) A 4-μm thick optical section of a 4-cell embryo (4cs). (d’) Higher magnification image of the box in (d). Arrow, the basal surface of a blastomere. (e) Stacked image of a whole 16-cell stage embryo (16cs). (e’) A 3-μm thick optical section at the vegetal hemisphere. Arrow, the basal surface of a blastomere at the vegetal hemisphere. (e”) A 3-μm thick optical section at the animal hemisphere. Arrow, the basal surface of a blastomere. (f) Stacked image of a whole 32-cell embryos (32cs). (f’) A 3-μm thick optical section at the equator region. Arrow, the blastomere basal surface. (g) A 2-μm optical section of swimming blastula (sBl). (g’) Higher magnification image of the box in (g). (g”) A 6-μm thick sagittal Polywax section. (h) A 2-μm thick optical section of a mesenchyme blastula (mBl). (h’) Higher magnification image of the box in (h). Arrow, primary mesenchyme cells. (i) A 3-μm thick optical section of early mesenchyme blastula (eG). (i’) Higher magnification image of the box in (i). Arrow, primary mesenchyme cells. (j) A 3-μm thick optical section of late gastrula (lG). (j’) Higher magnification image of the box in (j). Arrows, secondary mesenchyme cells. Scale bars, 20 μm (a–j’), 50 μm (g”).