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Chronic Exposure to Altered Gravity During the Pregnancy-to-Lactation Transition Affects Abundance of Cytoskeletal Proteins in the Rat Mammary Gland

Kibrom M. Alula
Kibrom M. Alula
, 
James H. Resau
James H. Resau
 and 
Osman V. Patel
Osman V. Patel
   | Jul 20, 2020
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Article Category: Research Article
Published Online: Jul 20, 2020
Page range: 58 - 72
DOI: https://doi.org/10.2478/gsr-2018-0005
Keywords
© 2018 Kibrom M. Alula et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Representative images of lobular outlining and unmixing for protein quantification. (a) An overlay image. (b) Overlay image unmixed into blue hematoxylin. (c) Overlay image unmixed into red cytokeratin. (d) Overlay image unmixed into brown vimentin. The unmixed images of a multi-labeled slide enable quantification of individual, as well as overlapping signals within the same outlined lobular border. The scale bar represents 50 μm (10X magnification) in the photomicrographs.
Representative images of lobular outlining and unmixing for protein quantification. (a) An overlay image. (b) Overlay image unmixed into blue hematoxylin. (c) Overlay image unmixed into red cytokeratin. (d) Overlay image unmixed into brown vimentin. The unmixed images of a multi-labeled slide enable quantification of individual, as well as overlapping signals within the same outlined lobular border. The scale bar represents 50 μm (10X magnification) in the photomicrographs.

Figure 2

Representative immunohistochemistry (IHC) sections of rat mammary gland from stationary-control (SC) and hypergravity-exposed (HG) animals at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3), stained for actin (red) and tubulin (brown). (a) Photomicrographs at 10X magnification with scale bar representing 50 μm. Boxes represent images at 40X. (b) Photomicrographs at 40X magnifications with scale bar representing 25 μm and arrows point to actin stained in red.
Representative immunohistochemistry (IHC) sections of rat mammary gland from stationary-control (SC) and hypergravity-exposed (HG) animals at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3), stained for actin (red) and tubulin (brown). (a) Photomicrographs at 10X magnification with scale bar representing 50 μm. Boxes represent images at 40X. (b) Photomicrographs at 40X magnifications with scale bar representing 25 μm and arrows point to actin stained in red.

Figure 3

Quantitative comparison of mammary gland lobular actin and tubulin proteins measured in stationary-control (SC) and hypergravity-exposed (HG) rat groups following unmixing of chromophores using the CRi Nuance Multispectral Imaging System at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3). (a) Comparison of actin levels within groups. (b) Comparison of actin levels between groups. (c) Comparison of tubulin levels within groups. (d) Comparison of tubulin levels between groups. Data are shown as Mean ± SEM; *means within and between each group are significantly different (*p<0.01; **p<0.005; ****p<0.0001). The scale has been expanded in insert Figure 3a and 3b to display the relationship of the quantified actin protein.
Quantitative comparison of mammary gland lobular actin and tubulin proteins measured in stationary-control (SC) and hypergravity-exposed (HG) rat groups following unmixing of chromophores using the CRi Nuance Multispectral Imaging System at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3). (a) Comparison of actin levels within groups. (b) Comparison of actin levels between groups. (c) Comparison of tubulin levels within groups. (d) Comparison of tubulin levels between groups. Data are shown as Mean ± SEM; *means within and between each group are significantly different (*p<0.01; **p<0.005; ****p<0.0001). The scale has been expanded in insert Figure 3a and 3b to display the relationship of the quantified actin protein.

Figure 4

Representative immunohistochemistry (IHC) sections of rat mammary gland from stationary-control (SC) and hypergravity-exposed (HG) animals at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3), stained for cytokeratin (red) and vimentin (brown). (a) Photomicrographs at 10X magnification with scale bar representing 50 μm. Boxes represent images at 40X. (b) Photomicrographs at 40X magnifications with scale bar representing 25 μm and arrows point to cytokeratin stained in red.
Representative immunohistochemistry (IHC) sections of rat mammary gland from stationary-control (SC) and hypergravity-exposed (HG) animals at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3), stained for cytokeratin (red) and vimentin (brown). (a) Photomicrographs at 10X magnification with scale bar representing 50 μm. Boxes represent images at 40X. (b) Photomicrographs at 40X magnifications with scale bar representing 25 μm and arrows point to cytokeratin stained in red.

Figure 5

Quantitative comparison of mammary gland lobular cytokeratin and vimentin proteins measured in stationary-control (SC) and hypergravity-exposed (HG) rat groups following unmixing of chromophores using the CRi Nuance Multispectral Imaging System at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3). (a) Comparison of cytokeratin levels within groups. (b) Comparison of cytokeratin levels between groups. (c) Comparison of vimentin levels within groups. (d) Comparison of vimentin levels between groups. Data are shown as Mean ± SEM; *means within and between each group are significantly different (*p<0.05; **p<0.005; ***p<0.001; ****p<0.0001). The scale has been expanded in insert Figure 5a and 5b to display the relationship of the quantified cytokeratin protein.
Quantitative comparison of mammary gland lobular cytokeratin and vimentin proteins measured in stationary-control (SC) and hypergravity-exposed (HG) rat groups following unmixing of chromophores using the CRi Nuance Multispectral Imaging System at day 20 of gestation (G20), postpartum day 1 (P1), and postpartum day 3 (P3). (a) Comparison of cytokeratin levels within groups. (b) Comparison of cytokeratin levels between groups. (c) Comparison of vimentin levels within groups. (d) Comparison of vimentin levels between groups. Data are shown as Mean ± SEM; *means within and between each group are significantly different (*p<0.05; **p<0.005; ***p<0.001; ****p<0.0001). The scale has been expanded in insert Figure 5a and 5b to display the relationship of the quantified cytokeratin protein.
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