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The SH2 domain is crucial for function of Fyn in neuronal migration and cortical lamination.

Lu X, Hu X, Song L, An L, Duan M, Chen S, Zhao S - BMB Rep (2015)

Bottom Line: Here, we demonstrate that the SH2 domain is essential for the action of Fyn in neuronal migration and cortical lamination.A point mutation in the Fyn SH2 domain (FynR176A) impaired neuronal migration and their final location in the cerebral cortex, by inducing neuronal aggregation and branching.Thus, we provide the first evidence of the Fyn SH2 domain contributing to neuronal migration and neuronal morphogenesis.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China.

ABSTRACT
Neurons in the developing brain form the cortical plate (CP) in an inside-out manner, in which the late-born neurons are located more superficially than the early-born neurons. Fyn, a member of the Src family kinases, plays an important role in neuronal migration by binding to many substrates. However, the role of the Src-homology 2 (SH2) domain in function of Fyn in neuronal migration remains poorly understood. Here, we demonstrate that the SH2 domain is essential for the action of Fyn in neuronal migration and cortical lamination. A point mutation in the Fyn SH2 domain (FynR176A) impaired neuronal migration and their final location in the cerebral cortex, by inducing neuronal aggregation and branching. Thus, we provide the first evidence of the Fyn SH2 domain contributing to neuronal migration and neuronal morphogenesis.

Show MeSH
FynR176A induced reorganization of F-actin and focal accumulation of vinculin CHO cells were cultured in F12K medium containing 10% FBS and transfected with GFP or FynR176A . (A, B) CHO cells were stained with phalloidin and DAPI, FynR176A induced the depolymerization of F-actin. (C, D) CHO cells were stained with tubulin; no difference was observed between the GFP group and FynR176A group. (E, F) CHO cells were stained with vinculin, and FynR176A partially colocalized with vinculin and induced focal accumulation of vinculin. (G) Western blotting analysis of vinculin, tubulin, and actin in the GFP group and FynR176A group; no difference was observed between these two groups. Scale bar = 20 μm
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Figure 004: FynR176A induced reorganization of F-actin and focal accumulation of vinculin CHO cells were cultured in F12K medium containing 10% FBS and transfected with GFP or FynR176A . (A, B) CHO cells were stained with phalloidin and DAPI, FynR176A induced the depolymerization of F-actin. (C, D) CHO cells were stained with tubulin; no difference was observed between the GFP group and FynR176A group. (E, F) CHO cells were stained with vinculin, and FynR176A partially colocalized with vinculin and induced focal accumulation of vinculin. (G) Western blotting analysis of vinculin, tubulin, and actin in the GFP group and FynR176A group; no difference was observed between these two groups. Scale bar = 20 μm

Mentions: We expressed FynR176A in CHO cells and stained cytoskeletal components, microtubules and F-actin, as well as the focal adhesion molecule, vinculin. We also transfected 293T cells for Western blot analysis. As shown in Fig. 4G, the expression of FynR176A did not affect the total amount of tubulin, actin, or vinculin. Immunochemistry results showed that FynR176A induced the depolymerization of F-actin. Well-organized F-actin was detected in the control group, while very little typical F-actin was found in the FynR176A group (Fig. 4). Expression of FynR176A also induced focal accumulation of the adhesion molecule, vinculin. As shown in Fig. 4, FynR176A partly colocalized with bundled vinculin, while no ‘typical’ bundled vinculin was observed in the control group (Fig. 4E). The expression and organization of another cytoskeletal component, microtubules, was not affected by the expression of FynR176A . Thus, we further demonstrated the mechanism by which FynR176A controls neuronal aggregation, adhesion, and branching.


The SH2 domain is crucial for function of Fyn in neuronal migration and cortical lamination.

Lu X, Hu X, Song L, An L, Duan M, Chen S, Zhao S - BMB Rep (2015)

FynR176A induced reorganization of F-actin and focal accumulation of vinculin CHO cells were cultured in F12K medium containing 10% FBS and transfected with GFP or FynR176A . (A, B) CHO cells were stained with phalloidin and DAPI, FynR176A induced the depolymerization of F-actin. (C, D) CHO cells were stained with tubulin; no difference was observed between the GFP group and FynR176A group. (E, F) CHO cells were stained with vinculin, and FynR176A partially colocalized with vinculin and induced focal accumulation of vinculin. (G) Western blotting analysis of vinculin, tubulin, and actin in the GFP group and FynR176A group; no difference was observed between these two groups. Scale bar = 20 μm
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4352619&req=5

Figure 004: FynR176A induced reorganization of F-actin and focal accumulation of vinculin CHO cells were cultured in F12K medium containing 10% FBS and transfected with GFP or FynR176A . (A, B) CHO cells were stained with phalloidin and DAPI, FynR176A induced the depolymerization of F-actin. (C, D) CHO cells were stained with tubulin; no difference was observed between the GFP group and FynR176A group. (E, F) CHO cells were stained with vinculin, and FynR176A partially colocalized with vinculin and induced focal accumulation of vinculin. (G) Western blotting analysis of vinculin, tubulin, and actin in the GFP group and FynR176A group; no difference was observed between these two groups. Scale bar = 20 μm
Mentions: We expressed FynR176A in CHO cells and stained cytoskeletal components, microtubules and F-actin, as well as the focal adhesion molecule, vinculin. We also transfected 293T cells for Western blot analysis. As shown in Fig. 4G, the expression of FynR176A did not affect the total amount of tubulin, actin, or vinculin. Immunochemistry results showed that FynR176A induced the depolymerization of F-actin. Well-organized F-actin was detected in the control group, while very little typical F-actin was found in the FynR176A group (Fig. 4). Expression of FynR176A also induced focal accumulation of the adhesion molecule, vinculin. As shown in Fig. 4, FynR176A partly colocalized with bundled vinculin, while no ‘typical’ bundled vinculin was observed in the control group (Fig. 4E). The expression and organization of another cytoskeletal component, microtubules, was not affected by the expression of FynR176A . Thus, we further demonstrated the mechanism by which FynR176A controls neuronal aggregation, adhesion, and branching.

Bottom Line: Here, we demonstrate that the SH2 domain is essential for the action of Fyn in neuronal migration and cortical lamination.A point mutation in the Fyn SH2 domain (FynR176A) impaired neuronal migration and their final location in the cerebral cortex, by inducing neuronal aggregation and branching.Thus, we provide the first evidence of the Fyn SH2 domain contributing to neuronal migration and neuronal morphogenesis.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China.

ABSTRACT
Neurons in the developing brain form the cortical plate (CP) in an inside-out manner, in which the late-born neurons are located more superficially than the early-born neurons. Fyn, a member of the Src family kinases, plays an important role in neuronal migration by binding to many substrates. However, the role of the Src-homology 2 (SH2) domain in function of Fyn in neuronal migration remains poorly understood. Here, we demonstrate that the SH2 domain is essential for the action of Fyn in neuronal migration and cortical lamination. A point mutation in the Fyn SH2 domain (FynR176A) impaired neuronal migration and their final location in the cerebral cortex, by inducing neuronal aggregation and branching. Thus, we provide the first evidence of the Fyn SH2 domain contributing to neuronal migration and neuronal morphogenesis.

Show MeSH