18 research outputs found
The number of genes expressed in respective regions.
<p>The number of genes expressed in respective regions.</p
AERO images of <i>Pax1</i> and <i>Aes.</i>
<p>Expression of <i>Pax1</i> (paired box gene 1) and <i>Aes</i> (amino-terminal enhancer of split) in the E11.5 mouse embryo was visualized by in situ hybridization. (A) Left- and Right-side AERO images of <i>Pax1</i>. The expression in the somite and maxillary process is observed. (B)Three AERO images of <i>Aes</i>. The Left-, Front- and Back-side AERO images cover almost all the body parts of the embryo. The expression in the brain, somite, forelimb and hindlimb is clearly observed from various angles.</p
Comparison of conventional 2D images and AERO images.
<p>The expression of <i>Lmx1b</i>, <i>Isl1</i> and <i>Cited1</i> at E11.5 is visualized by in situ hybridization. The conventional 2D images (A, C, E) from the EMBRYS database and the AERO images (B, D, F). (A, B) The AERO images reveal <i>Lmx1b</i> expression is restricted to the dorsal side and is absent on the ventral side of the forelimb-bud (arrow) and hindlimb bud (arrow head). D:dorsal side, V: ventral side. (C, D) <i>Isl1</i> expression in the genitalia is clearly observed in the AERO images (red arrows). (E, F) <i>Cited1</i> expression in the maxillofacial region is more clearly observed in the AERO images than 2D image (red arrows).</p
Overall setup of the AERO system hardware.
<p>(A) HiRox microscopy and (B) schematic representation of the rotary head. A specimen is placed on an agarose layer prepared in a cylindrical container or petri-dish. The key feature of this component is the two mirrors installed in the rotary head, which allows the camera to capture an image that is tilted 45 degrees to the microscopic stage. The rotary head is rotated by 360 degrees while capturing images at 2-degree intervals. Therefore the 180 serial images are captured in one simple operation. After image acquisition, these serial images are automatically arranged so as to produce a movie.</p
The AERO System: A 3D-Like Approach for Recording Gene Expression Patterns in the Whole Mouse Embryo
<div><p>We have recently constructed a web-based database of gene expression in the mouse whole embryo, EMBRYS (<a href="http://embrys.jp/embrys/html/MainMenu.html" target="_blank">http://embrys.jp/embrys/html/MainMenu.html</a>). To allow examination of gene expression patterns to the fullest extent possible, this database provides both photo images and annotation data. However, since embryos develop via an intricate process of morphogenesis, it would be of great value to track embryonic gene expression from a three dimensional perspective. In fact, several methods have been developed to achieve this goal, but highly laborious procedures and specific operational skills are generally required. We utilized a novel microscopic technique that enables the easy capture of rotational, 3D-like images of the whole embryo. In this method, a rotary head equipped with two mirrors that are designed to obtain an image tilted at 45 degrees to the microscope stage captures serial images at 2-degree intervals. By a simple operation, 180 images are automatically collected. These 2D images obtained at multiple angles are then used to reconstruct 3D-like images, termed AERO images. By means of this system, over 800 AERO images of 191 gene expression patterns were captured. These images can be easily rotated on the computer screen using the EMBRYS database so that researchers can view an entire embryo by a virtual viewing on a computer screen in an unbiased or non-predetermined manner. The advantages afforded by this approach make it especially useful for generating data viewed in public databases.</p></div
Enlargement of GLA due to slow cell proliferation and inter-gla fusion in the 3d stemness-inducing nanoenvironment.
<p>(A) Representative photomicrographs of PC-3 cells after reaching confluent. Cells were cultured in the 2D condition. Cellular morphologies at day 4, 5, and 7 were shown. Arrowheads indicate GLA on the 2D monolayer cells. Scale bar, 100 μm. (B) Representative photomicrographs of PC-3 cells in the 3D culture condition. Cellular morphologies at day 11 and 14 were shown. Scale bar, 100 μm. (C) Growth curves of PC-3 cells cultured in 2D serum-contained and 3D stem cell medium conditions. Cells were cultured in a 96-well plate. **P < 0.01 (2D serum vs 3D stem), n = 3. (D) Growth curves of PC-3 cells cultured in 2D serum-contained and 2D stem cell medium conditions. *P < 0.05 (2D stem vs 2D serum), n = 3. (E) Growth curves of PC-3 cells cultured in 3D serum-contained and 3D stem cell medium conditions. *P < 0.05 (3D stem vs 3D serum), n = 3. **P < 0.01 (3D stem vs 3D serum), n = 3. (F-H) Viabilities of PC-3 cells cultured in 2D or 3D conditions in serum-contained or stem cell media. Same data with different vertical axis values were shown between F and H and between G and I. (F, H) P < 0.05 (2D serum vs 2D stem), n = 3. (G, I) *P < 0.05 (vs day 0), n = 3. **P < 0.01 (vs day 0), n = 3. 3D serum d0 vs d7, P = 0.028. 3D serum d0 vs d11, P = 0.0052. 3D serum d0 vs d14, P = 0.0012. 3D stem d0 vs d7, P = 0.0138. 3D stem d0 vs d11, P = 0.0007. 3D stem d0 vs d14, P = 0.0004.</p
Classification of the morphologies of cellular aggregations.
<p>Classification of the morphologies of cellular aggregations.</p
Tumor morphologies and metastatic potentials of the prostate-derived cell lines.
<p>Tumor morphologies and metastatic potentials of the prostate-derived cell lines.</p
Gene expression switching of Epithelial-Splicing Regulatory Proteins (ESRPs), CD44 variant, and stem cell markers depending on cell culture nanoEnvironments.
<p>(A) Representative morphologies of PC-3 cells cultured in the 4 different conditions. Cells were cultured in 10% serum-containing F12K medium or mTeSR1 stem-cell medium on 2D plates or 3D NCPs. Arrows indicate projections of cells. Scale bars, 100 μm. (B) Schematic structures of <i>CD44</i> gene, <i>CD44 variant 8–10</i> (<i>CD44v8-10</i>) and <i>CD44 standard</i> (<i>CD44s</i>). Blue and gray rectangles represent standard exons (exon 1 to 10) and variant exons (V1 to V10), respectively. The red primer pair is for all variants and the CD44s. The green primer pair is for CD44v containing exon V9. The blue primer pair is for CD44s only. (C) Agarose gel electrophoresis analysis of RT-PCR amplicons of CD44v and CD44s. An arrow indicates <i>CD44v8-10</i> amplicon. An arrowhead indicates <i>CD44s</i> amplicon. M1k, a 1 kbp DNA ladder marker. M100, a 100 bp DNA ladder marker. <i>ACTB</i>, β-actin mRNA as an internal control. (D) qRT-PCR analysis of stem-cell-related and epithelial-splicing regulatory genes. The mRNA expression levels of <i>CD44s</i>, <i>CD44v</i>, <i>ECAD/CDH1</i>, <i>ESRP1</i>, <i>ESRP2</i>, and <i>CD133</i> were examined. Relative mRNA expression levels versus those of <i>GAPDH</i> are shown. n = 3. (E) Flow cytometry analysis of CD44v9. PC-3 cells were cultured in serum-containing medium and passage number 1, 7, and 13 were examined by flow cytometry. An anti-prostate-specific antigen (PSA) antibody was used as a negative control. Serum promoted differentiation of the cells and reduced stemness.</p