Journal: Scientific Reports

Article Title: Amyloid-β slows cilia movement along the ventricle, impairs fluid flow, and exacerbates its neurotoxicity in explant culture

doi: 10.1038/s41598-023-40742-0

Figure Lengend Snippet: Neurotoxic effects of Aβ1–42 observed in the explant culture. ( A ) Fluorescence images of tubulin βIII positive neurons migrated from the explant; images are from the area 0.5 mm up and down, left and right of the schematically illustrated explant. ( B ) Typical time-lapse images of neurites retracted in 10 µM Aβ1–42 (time-lapse images were taken at 1, 2, 3, 4, 5, and 13 h from left to right). The soma is shown by the arrows. ( C ) A typical shrunken neuron in 3 µM Aβ1–42 for three days is positive for tubulin βIII (left) and superimposed on the DIC image (right). Neurons in the control medium in the same notation (lower panels). ( D ) TB-positive shrunken cells in the explant culture in the Aβs. DIC images of explant culture of the control (upper), Aβ1–42 (3 µM, middle), and Aβ1–40 (3 µM, lower). Live (green) and dead (red) cell staining of neurons superimposed on the DIC images (10 µM Aβ1–42 for four days, bottom) on the cilia side (left column) and non-cilia side (right column). ( E ) A typical explant culture treated with Aβ1–42 (10 µM) and RAβ1–42 (0.4 µM). (a) DIC image of an explant culture. The yellow arrow shows a typical shrunken cell. (b) Fluorescence image of RAβ1–42. The shrunken cell positive for RAβ1–42 is pointed by the arrow. (c) Neurons on the ciliated side are positive for MAP-2 (blue). Small fraction of neurons positive for RAβ1–42 (magenta) shown by an arrow. (d) Neurons on the non-ciliated side are positive for MAP-2 and RAβ1–42. The green line shows the ependymal cilia cells (a and b). ( F ) (a)The distribution of RAβ1–42 plotted in the polar coordinate system of the explant culture; the fluorescence intensity of RAβ1–42 in the area 100 μm from the edge of the explant culture was plotted. N = 3 culture wells. The inset is an illustration of an explant culture and RAβ1–42 positive cells (red dots) and the assignment of the angle; the polar coordinate system of the explant culture is divided into 12 sections [ 0, 30), [ 30, 60) …, [ 330, 360); the center of the ciliated area is assigned 180 degrees. (b) The individual bar denotes the distribution of beating cilia. The fluorescence intensity of RAβ1–42 on the non-ciliated area is significantly higher than that on the ciliated area control ( p = 0.006, one-way ANOVA test, Origin ver. 2020b). ( G ) fluorescence image of RAβ1–42 positive cells of an explant culture that had no ciliated cells. Bars are 50 µm (panel a) and 30 µm (panels C ), 100 µm (panels B and D ), 500 µm (panels E a and b), 50 µm (panels E c and d), and 300 µm (panel G ). N denotes the number of culture wells.

Article Snippet: For immunocytochemistry, cells were fixed with 4% paraformaldehyde, and incubated overnight at 4 °C with mouse anti-β-III-tubulin (1:100; Promega, USA) and with donkey anti-mouse IgG 488 (1:100; FluoProbes, USA), which showed the majority of large soma with neurites were neurons.

Techniques: Fluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Generation of Periventricular Reactive Astrocytes Overexpressing Aquaporin 4 Is Stimulated by Mesenchymal Stem Cell Therapy

doi: 10.3390/ijms24065640

Figure Lengend Snippet: Integration of BM-MSCs into the periventricular cerebral tissue of hydrocephalic hyh mice. ( a ) Wall of the neocortical (ncx) lateral ventricle (v) of a hydrocephalic hyh mouse at P18; administered BM-MSCs (white arrows, mRFP1 fluorescence in red) at P4. BM-MSCs are located between periventricular reactive astrocytes (GFAP, green). ( b , b′ ) Separated channels showing fluorescence of the green cell tracker and mRFP1 (red) in the transplanted BM-MSCs. ( c , c′ , c″ ) Labeling of BM-MSCs (arrow, mRFP1 fluorescence in red) with an antibody against mRFP1 (green, white arrow). Expression of ( d , d′ , d″ ) δGFAP (green), ( e , e′ , e″ ) nestin (green), ( f – f″ ) NG2 (green), ( g – g″ ) βIII-tubulin (green), and ( h – h″ ) GFAP (green) in BM-MSCs (mRFP1, fluorescence in red, arrows). ( f’ , f″ , g′ , g″ ) are separated channels for fluorescence of the cells indicated in the framed area in f and g , respectively. The BM-MSCs present a weak immunoreaction (arrows) compared to NG2 cells and reactive astrocytes of the host tissue (arrowheads in ( f , f′ ), arrows in ( h – h″ )). Magnifications are the same in the different separated channels. All images are from immunofluorescence in vibratome sections. mRNA levels of the interleukins ( i ) IL-1α and ( j ) IL-1β, and ( k ) CD45. Means ± SEM are shown (nonhydrocephalic mice, nh; = 5; hyh hydrocephalic sham controls, sham, n = 6, n = 4 for CD45; hyh hydrocephalic treated with BM-MSCs, BM-MSC, n = 7, n = 5 for CD45). * p < 0.05, ** p < 0.01; Wilcoxon–Mann–Whitney test. Abbreviations: str, striatum.

Article Snippet: β–III tubulin , Promega, A6712 , Mouse monoclonal , 1:5000, I.

Techniques: Fluorescence, Labeling, Expressing, Immunofluorescence, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Generation of Periventricular Reactive Astrocytes Overexpressing Aquaporin 4 Is Stimulated by Mesenchymal Stem Cell Therapy

doi: 10.3390/ijms24065640

Figure Lengend Snippet: Upregulation of proteins associated with the overexpressed pathways in hyh mice 14 days after BM-MSC treatment at P4. Sum PEP score corresponds to the score calculated based on the posterior error probability (PEP) values of the peptide spectrum matches (PSM). Sum PEP score indicates the probability that an observed PSM is incorrect. Abundance ratio (AR) indicates the relation of the protein expression in BM-MSC-transplanted mice/sham-injected hyh mice.

Article Snippet: β–III tubulin , Promega, A6712 , Mouse monoclonal , 1:5000, I.

Techniques: Expressing, Protein Binding, Migration

Journal: International Journal of Molecular Sciences

Article Title: Generation of Periventricular Reactive Astrocytes Overexpressing Aquaporin 4 Is Stimulated by Mesenchymal Stem Cell Therapy

doi: 10.3390/ijms24065640

Figure Lengend Snippet: Primary antibodies used for immunolabeling. Chromotek; Planegg-Martinsried, Germany. DSHB; Developmental Studies Hybridoma Bank, Iowa City, IA, USA. Promega; Madison, WI, USA. Abbreviations: I, immunofluorescence or immunohistochemistry; WB, Western blot.

Article Snippet: β–III tubulin , Promega, A6712 , Mouse monoclonal , 1:5000, I.

Techniques: Immunolabeling, Immunofluorescence, Immunohistochemistry, Western Blot, In Vitro

Journal: ACS Biomaterials Science & Engineering

Article Title: Aligned Polyhydroxyalkanoate Blend Electrospun Fibers as Intraluminal Guidance Scaffolds for Peripheral Nerve Repair

doi: 10.1021/acsbiomaterials.2c00964

Figure Lengend Snippet: Confocal micrographs of NG108-15 neuronal cells immunolabeled for βIII-tubulin and DAPI on (A) P(3HO)/P(3HB) (50:50) 5 μm fibers, (B) P(3HO)/P(3HB) (50:50) 8 μm fibers, (C) PCL 5 μm fibers, (D) PCL 8 μm fibers, and (E) tissue culture plastic control (scale bar = 100 μm). (F) The percentage of neurite bearing neuronal cells after 6 days in culture. (G) The average number of neurites expressed per neuron. (H) Average neurite length per condition after 6 days in culture (mean ± SD, n = 3 independent experiments; * p < 0.05 and ** p < 0.01).

Article Snippet: NG108-15 neuronal cells, and neurites outgrown from DRG explants, were labeled for βIII-tubulin (neurite marker) using a mouse anti-β III-tubulin antibody (1:500 dilution in 1% BSA) (Promega, UK) and incubated for 24 h at 4 ° C. After a wash in PBS, a Texas Red-conjugated anti-mouse IgG antibody was added to samples (1:100 dilution in 1% BSA) (Vector Labs, USA) for 2 h at room temperature.

Techniques: Immunolabeling

Journal: ACS Biomaterials Science & Engineering

Article Title: Aligned Polyhydroxyalkanoate Blend Electrospun Fibers as Intraluminal Guidance Scaffolds for Peripheral Nerve Repair

doi: 10.1021/acsbiomaterials.2c00964

Figure Lengend Snippet: Confocal micrographs DRG explants on fibers immunolabeled for βIII tubulin, S100β, and DAPI on (A) P(3HO)/P(3HB) (50:50) 5 μm fibers, (B) P(3HO)/P(3HB) (50:50) 8 μm fibers, (C) PCL 5 μm fibers, and (D) PCL 8 μm fibers (scale bar = 200 μm). (E) Schwann cell migration length and (F) average neurite outgrowth length were determined from confocal micrographs (mean ± SD, n = 3 independent experiments; * p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: NG108-15 neuronal cells, and neurites outgrown from DRG explants, were labeled for βIII-tubulin (neurite marker) using a mouse anti-β III-tubulin antibody (1:500 dilution in 1% BSA) (Promega, UK) and incubated for 24 h at 4 ° C. After a wash in PBS, a Texas Red-conjugated anti-mouse IgG antibody was added to samples (1:100 dilution in 1% BSA) (Vector Labs, USA) for 2 h at room temperature.

Techniques: Immunolabeling, Migration

Journal: Neurology Research International

Article Title: Differentiation of Human Breast-Milk Stem Cells to Neural Stem Cells and Neurons

doi: 10.1155/2014/807896

Figure Lengend Snippet: (a) Neurons and astrocytes differentiated from breast milk stem cells-derived neural stem cell stained with anti- β -tubulin-Alexa Flour 568 (red) and anti-GFAP-Alexa Flour 488 (green); (b) oligodendrocyte differentiated from breast milk stem cell-derived neural stem cell stained with anti-O4-Alexa Flour 568.

Article Snippet: For neuron, oligodendrocyte, and astrocyte detection, antibodies, β -tubulin III (Promega G7121, 1 : 2000), O4 [ ] (Millipore, MAB345 1 : 50), and GFAP [ ] (DakoCytomation, Code number Z0334 1 : 500), respectively, were assessed by an immunocytochemistry method similar to the one previously described.

Techniques: Derivative Assay, Staining

Journal: Neurology Research International

Article Title: Differentiation of Human Breast-Milk Stem Cells to Neural Stem Cells and Neurons

doi: 10.1155/2014/807896

Figure Lengend Snippet: The percentage of β -tubulin, O4, and GFAP.

Article Snippet: For neuron, oligodendrocyte, and astrocyte detection, antibodies, β -tubulin III (Promega G7121, 1 : 2000), O4 [ ] (Millipore, MAB345 1 : 50), and GFAP [ ] (DakoCytomation, Code number Z0334 1 : 500), respectively, were assessed by an immunocytochemistry method similar to the one previously described.

Techniques: