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Neural cell adhesion molecule (NCAM) and polysialic acid–NCAM expression in developing ICR mice

Chairat Turbpaiboon
Chairat Turbpaiboon
, 
Wongsakorn Siripan
Wongsakorn Siripan
, 
Pornkanok Nimnoi
Pornkanok Nimnoi
, 
Gopinathan Pillai Sreekanth
Gopinathan Pillai Sreekanth
, 
Witthawat Wiriyarat
Witthawat Wiriyarat
, 
Boonrat Tassaneetrithep
Boonrat Tassaneetrithep
 and 
Supin Chompoopong
Supin Chompoopong
   | Sep 25, 2019
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Article Category: Brief communication (original)
Published Online: Sep 25, 2019
Page range: 179 - 187
DOI: https://doi.org/10.1515/abm-2019-0018
Keywords
© 2018 Chairat Turbpaiboon et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

E17 fetal and P3, P5, and P7 postnatal mice with normal appearance. The black scale bar for E17 represents 1 mm. Scale bars in white for the pups represent 1 cm. Neural cell adhesion molecule (NCAM) immunoreactivity (brown) and hematoxylin (purple)-stained coronal brain sections were also compared. Progression of neocortex shows 6 cortical cell layers starting from E17 to P7. At E17, cortical neurons continue to aggregate and separate into layers, but postnatal brains P3, P5, and P7 show the migration of cortical neurons and their location in overlapped layers. NCAM immunoreactivity was increased from E17 to P7. The double-ended arrow indicates the measurement of the cortical thickness. Scale bars on the coronal sections represent 200 μm E17, fetal mouse at embryonic age of 17 days; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively
E17 fetal and P3, P5, and P7 postnatal mice with normal appearance. The black scale bar for E17 represents 1 mm. Scale bars in white for the pups represent 1 cm. Neural cell adhesion molecule (NCAM) immunoreactivity (brown) and hematoxylin (purple)-stained coronal brain sections were also compared. Progression of neocortex shows 6 cortical cell layers starting from E17 to P7. At E17, cortical neurons continue to aggregate and separate into layers, but postnatal brains P3, P5, and P7 show the migration of cortical neurons and their location in overlapped layers. NCAM immunoreactivity was increased from E17 to P7. The double-ended arrow indicates the measurement of the cortical thickness. Scale bars on the coronal sections represent 200 μm E17, fetal mouse at embryonic age of 17 days; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively

Figure 2

Coronal section through the hippocampus of P3 postnatal mice indicating layers in the DG: po layer, strata gr, and mo. Immunoperoxidase staining with NCAM (A) was localized to the extracellular matrix around cell surface of neurons. *Nonstaining neuronal cells and SGZ indicated by arrows for neuronal lining at the base of gr layer. PSA–NCAM (B) was localized in the cytoplasm and processes of neural cells (indicated by arrow heads). Scale bar represents 50 μm DG, dentate gyrus; gr, stratum granulosum; mo, stratum moleculare; po, polymorphic; SGZ, subgranular zone
Coronal section through the hippocampus of P3 postnatal mice indicating layers in the DG: po layer, strata gr, and mo. Immunoperoxidase staining with NCAM (A) was localized to the extracellular matrix around cell surface of neurons. *Nonstaining neuronal cells and SGZ indicated by arrows for neuronal lining at the base of gr layer. PSA–NCAM (B) was localized in the cytoplasm and processes of neural cells (indicated by arrow heads). Scale bar represents 50 μm DG, dentate gyrus; gr, stratum granulosum; mo, stratum moleculare; po, polymorphic; SGZ, subgranular zone

Figure 3

Coronal sections through DG of mice at postnatal day 5 (P5) (A and C) indicating layers in the DG: po layer, strata gr, and mo. PSA–NCAM immunoperoxidase staining with (A and B) or without (C and D) hematoxylin was localized in the cytoplasm and nucleus of neurons in gr (arrow heads) and intense staining in the small nucleus (arrows) is found in lm and ra layers of CA1. Scale bars are 100 μm (C), 50 μm (A), and 20 μm (B, D) CA1, cornu ammonis 1 area of the hippocampus; DG, dentate gyrus; gr, stratum granulosum; lm, stratum lacunosum moleculare; mo, stratum moleculare; NCAM, neural cell adhesion molecule; po, polymorphic layer; PSA, polysialic acid; ra, stratum radiatum
Coronal sections through DG of mice at postnatal day 5 (P5) (A and C) indicating layers in the DG: po layer, strata gr, and mo. PSA–NCAM immunoperoxidase staining with (A and B) or without (C and D) hematoxylin was localized in the cytoplasm and nucleus of neurons in gr (arrow heads) and intense staining in the small nucleus (arrows) is found in lm and ra layers of CA1. Scale bars are 100 μm (C), 50 μm (A), and 20 μm (B, D) CA1, cornu ammonis 1 area of the hippocampus; DG, dentate gyrus; gr, stratum granulosum; lm, stratum lacunosum moleculare; mo, stratum moleculare; NCAM, neural cell adhesion molecule; po, polymorphic layer; PSA, polysialic acid; ra, stratum radiatum

Figure 4

Coronal sections through hippocampus of E17 fetal mice (A and E), and P3 (B and F), P5 (C and G), and P7 (D and H) postnatal mice after immunoperoxidase staining for NCAM (A–D) and PSA–NCAM (E–H). Scale bars represent 200 μm E17, mouse at embryonic age of 17 days; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid
Coronal sections through hippocampus of E17 fetal mice (A and E), and P3 (B and F), P5 (C and G), and P7 (D and H) postnatal mice after immunoperoxidase staining for NCAM (A–D) and PSA–NCAM (E–H). Scale bars represent 200 μm E17, mouse at embryonic age of 17 days; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid

Figure 5

Image analysis of NCAM (A) and PSA–NCAM (B) immunoreactivity in the SGZ of dentate expressed as % area. The immunoreactivity in the brains of fetal E17 and postnatal P3, P5, and P7 mice was compared. Significant differences (*P < 0.05) were found at P3, P5, and P7 when compared with E17. Bars indicate mean % area and error bars indicate SEM E17, mouse at embryonic age of 17 days; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid; SEM, standard error of the mean; SGZ, subgranular zone
Image analysis of NCAM (A) and PSA–NCAM (B) immunoreactivity in the SGZ of dentate expressed as % area. The immunoreactivity in the brains of fetal E17 and postnatal P3, P5, and P7 mice was compared. Significant differences (*P < 0.05) were found at P3, P5, and P7 when compared with E17. Bars indicate mean % area and error bars indicate SEM E17, mouse at embryonic age of 17 days; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid; SEM, standard error of the mean; SGZ, subgranular zone

Figure 6

Western blot analysis of NCAM (A) and PSA–NCAM (B) immunoreactivity in brain tissue (homogenates) from individual P3, P5, and P7 postnatal mice (n = 3). Bars indicate mean values of NCAM and PSA–NCAM immunoreactivity levels normalized to that for GAPDH. Significant differences (*P < 0.05) were found for P5 and P7 when compared with P3. Error bars indicate SEM GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid; SEM, standard error of the mean
Western blot analysis of NCAM (A) and PSA–NCAM (B) immunoreactivity in brain tissue (homogenates) from individual P3, P5, and P7 postnatal mice (n = 3). Bars indicate mean values of NCAM and PSA–NCAM immunoreactivity levels normalized to that for GAPDH. Significant differences (*P < 0.05) were found for P5 and P7 when compared with P3. Error bars indicate SEM GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NCAM, neural cell adhesion molecule; P3, P5, and P7, postnatal mice 3, 5, and 7 days after birth respectively; PSA, polysialic acid; SEM, standard error of the mean

Cortical thickness, brain weight, and body weight, and NCAM expression in the cerebral cortex of E17 fetal, and P3, P5, and P7 postnatal mice

Variable (mean ± SD)E17P3P5P7
Cortical thickness (μm)541 ± 4.35574.14 ± 6.4*780.96 ± 7.05*
Brain weight (g)–0.21 ± 0.010.24 ± 0.02*0.27 ± 0.08*
Body weight (g)2.77 ± 0.143.28 ± 0.21*4.71 ± 0.41*
Body length (cm)5.58 ± 0.045.64 ± 0.07*7.39 ± 0.03*
NCAM expression in the cerebral cortex (integrated density)25.87 ± 2.46234.75 ± 13.61259.13 ± 11.71387.34 ± 5.87
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