Journal: Stem cell research

Article Title: EphrinB3 restricts endogenous neural stem cell migration after traumatic brain injury

doi: 10.1016/j.scr.2016.09.029

Figure Lengend Snippet: EphrinB3 and CCI injury alter neuroblast migration in the subventricular zone (SVZ). The SVZ was dissected from non-injured (a,b) and CCI injured (c,d) wild type (WT) (a,c) and ephrinB3−/− (b,d) mice, and labeled with anti-DCX (red) and anti-CD31 (green) antibodies. In non-injured WT, neuroblasts can be seen predominantly in thick streams (arrowheads) across the dorsal aspect of the SVZ (a). High magnification insets (a′,a″). CCI injury leads to a mixture of tight (arrowheads) and broader streams (arrows) (c). High magnification insets (c′,c″). In ephrinB3−/− mice, neuroblasts are mainly observed in broad streams (b) and little change after CCI injury (d). High magnification insets (b′,b″,d′,d″).

Article Snippet: Explants were cultured under four conditions: basal (Matrigel alone);Matrigel containing 1 μg/mL human anti-Fc-molecules (Vector Laboratories, USA); Matrigel containing 10 μg/mL ephrinB3-Fc (R&D Systems Inc., USA) or Matrigel containing clustered ephrinB3-Fc (10 μg/mL ephrinB3 pretreated with 1 μg/mL human anti-Fc molecules for 30 min at room temperature).

Techniques: Migration, Labeling

Journal: Stem cell research

Article Title: EphrinB3 restricts endogenous neural stem cell migration after traumatic brain injury

doi: 10.1016/j.scr.2016.09.029

Figure Lengend Snippet: EphrinB3 increases neuroblast migration outside the SVZ and RMS. Immunolabeled sagittal brain section from a WT mouse showing anti-DCX labeled neuroblasts (red) in RMS, CC and perilesional cortex 3 days after CCI injury, while anti-CD31 (green) labeled vessels were used for tissue referencing (a). Stereological cell counts of the RMS (b), rostral CC (c), caudal CC (d) and peri-lesional cortex (e) show a rostral-to-caudal gradient of neuroblasts from the rostral CC towards the caudal CC and injury site. Increased neuroblasts numbers were observed in the rostral CC (c), caudal CC (d) and perilesional cortex (e) in ephrinB3−/− as compared to WT mice. High-magnification images of neuroblasts in the rostral CC (f), caudal CC (g) and perilesional cortex (h). CC, corpus callosum; DG, dentate gyrus; Hipp, Hippocampus; LV, lateral ventricle; OB, olfactory bulb; RMS, rostral migratory stream; SVZ, subventricular zone. *p < 0.05, **p < 0.01 as compared with their respective non-injured controls.

Article Snippet: Explants were cultured under four conditions: basal (Matrigel alone);Matrigel containing 1 μg/mL human anti-Fc-molecules (Vector Laboratories, USA); Matrigel containing 10 μg/mL ephrinB3-Fc (R&D Systems Inc., USA) or Matrigel containing clustered ephrinB3-Fc (10 μg/mL ephrinB3 pretreated with 1 μg/mL human anti-Fc molecules for 30 min at room temperature).

Techniques: Migration, Immunolabeling, Labeling

Journal: Stem cell research

Article Title: EphrinB3 restricts endogenous neural stem cell migration after traumatic brain injury

doi: 10.1016/j.scr.2016.09.029

Figure Lengend Snippet: EphrinB3-Fc infusion reduces neuroblast migration into the overlying CC at 2 days following cannula injury. Immunolabeled sagittal brain section from a WT mouse shows anti-DCX labeled neuroblasts (red) in the RMS, CC and subcortical tissues at 2 days post-injury, while anti-CD31 (green) labeled vessels were used for tissue referencing (a). High-magnification inset of neuroblasts migrating from RMS to CC and subcortical tissues (arrowheads depict neuroblast chains) (a′). Stereology shows increased numbers of neuroblasts in the RMS (b) and CC (c) in Fc-control WT mice as compared with non-injured WT mice. Infusion of ephrinB3-Fc reduced neuroblast numbers in the CC of WT mice (c). EphrinB3−/− mice show increased chain (d) and isolated cell (e) migration in the rostral CC as compared with WT mice. Infusion of clustered ephrinB3-Fc reduced the number of neuroblast chains/clusters but increased the number of isolated neuroblasts. CC, corpus callosum; RMS, rostral migratory stream; SVZ, subventricular zone. *p < 0.05, **p < 0.01, ***p < 0.001 as compared with their respective non-injured controls; #p < 0.05 as compared with their respective Fc controls.

Article Snippet: Explants were cultured under four conditions: basal (Matrigel alone);Matrigel containing 1 μg/mL human anti-Fc-molecules (Vector Laboratories, USA); Matrigel containing 10 μg/mL ephrinB3-Fc (R&D Systems Inc., USA) or Matrigel containing clustered ephrinB3-Fc (10 μg/mL ephrinB3 pretreated with 1 μg/mL human anti-Fc molecules for 30 min at room temperature).

Techniques: Migration, Immunolabeling, Labeling, Isolation

Journal: Stem cell research

Article Title: EphrinB3 restricts endogenous neural stem cell migration after traumatic brain injury

doi: 10.1016/j.scr.2016.09.029

Figure Lengend Snippet: Application of ephrinB3 regulates neuroblast chain migration from cultured SVZ explants. Brightfield (a,c,e) and DCX-immunolabeled (b,d,f) SVZ explants show migrating neuroblast in basal (a,b) conditions, unclustered (Uncl.) ephrinB3-Fc (c,d) to block Eph signaling, and clustered (Cl.) ephrinB3-Fc (e,f) to activate Eph signaling. The extent of chain migration and single cell migration were measured using a semi-quantitative scale: 0 = no outgrowth; 1 ≤ 10 chains/cells; 2 = 10–50 chains/cells; 3 = 50–100 chains/cells; 4 = extensive growth. The area of explant outgrowth was measured and expressed as a ratio of outgrowth area (i.e. explant size). Graphs show increased numbers of chains migrating out of the explants after treatment with Uncl. ephrinB3-Fc (g), while isolated cells were increased after treating with Cl. ephrinB3-Fc (h). Both Cl. and Uncl. ephrinB3-Fc increased the total outgrowth of the explants (i). Results in all graphs show mean ± SEM of 39–48 explants per treatment; *p < 0.05 compared to basal conditions.

Article Snippet: Explants were cultured under four conditions: basal (Matrigel alone);Matrigel containing 1 μg/mL human anti-Fc-molecules (Vector Laboratories, USA); Matrigel containing 10 μg/mL ephrinB3-Fc (R&D Systems Inc., USA) or Matrigel containing clustered ephrinB3-Fc (10 μg/mL ephrinB3 pretreated with 1 μg/mL human anti-Fc molecules for 30 min at room temperature).

Techniques: Migration, Cell Culture, Immunolabeling, Blocking Assay, Isolation

Journal: Stem cell research

Article Title: EphrinB3 restricts endogenous neural stem cell migration after traumatic brain injury

doi: 10.1016/j.scr.2016.09.029

Figure Lengend Snippet: EphrinB3 is expressed in the human control and TBI brains. Expression of ephrinB3 in uninjured (n = 2) or traumatized (n = 7) human brain samples as detected by Western blotting. The traumatized samples were grouped into tissue collection between 3 and 24 h (acute; n= 6) or >5 days (122 h; n = 1). No differences were observed between acute TBI samples and uninjured controls; however, in the one sample collected from a patient 122 h after injury a reduced level of ephrinB3 was observed despite a high level of expression of β-actin in this sample.

Article Snippet: Explants were cultured under four conditions: basal (Matrigel alone);Matrigel containing 1 μg/mL human anti-Fc-molecules (Vector Laboratories, USA); Matrigel containing 10 μg/mL ephrinB3-Fc (R&D Systems Inc., USA) or Matrigel containing clustered ephrinB3-Fc (10 μg/mL ephrinB3 pretreated with 1 μg/mL human anti-Fc molecules for 30 min at room temperature).

Techniques: Expressing, Western Blot