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Essential roles of leucine-rich glioma inactivated 1 in the development of embryonic and postnatal cerebellum.

Xie YJ, Zhou L, Jiang N, Zhang N, Zou N, Zhou L, Wang Y, Cowell JK, Shen Y - Sci Rep (2015)

Bottom Line: The function of LGI1 in CNS development remains undefined.BrdU staining showed an inhibited proliferation of CGPs in KO embryos, which might be explained by the reduced Sonic hedgehog in embryos.Together, our results demonstrate that LGI1 is an essential player in the cerebellar development.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.

ABSTRACT
Leucine-rich glioma inactivated 1 (LGI1) is a secreted protein that interacts with ADAM transmembrane proteins, and its mutations are linked to human epilepsy. The function of LGI1 in CNS development remains undefined. Here, we report novel functions of LGI1 in the generation of cerebellar granule precursors (CGPs) and differentiation of radial glial cells (RGCs) in the cerebellum. A reduction in external granule layer thickness and defects in foliation were seen in embryonic and new-born LGI1 knockout (KO) mice. BrdU staining showed an inhibited proliferation of CGPs in KO embryos, which might be explained by the reduced Sonic hedgehog in embryos. In addition, the differentiation of RGCs into Bergmann glias was suppressed in KO mice. Enhanced Jagged1-Notch1 signaling in KO mice via reduced β-secretase proteolysis suggests that altered phenotype of RGCs is due to abnormal Notch1 signaling. Together, our results demonstrate that LGI1 is an essential player in the cerebellar development.

No MeSH data available.


Related in: MedlinePlus

Nestin expression is normal in the KO mice.(a) Nestin staining in the cerebellum (E15.5) and cerebellar lobules (P1). Arrowheads indicate abnormal EGL thickness and folia formation (n = 7 pairs for each age). Scale bars: 50 μm. (b) Total nestin expression levels were measured by western blotting using GAPDH as the internal control. (c) Quantitation of nestin immunoreactivity. The nestin/GAPDH ratios were 61.7 ± 8.0% (WT, E15.5) and 60.6 ± 6.9% (KO, E15.5) (n = 3, p = 0.34); 62.6 ± 8.0% (WT, P1) and 64.1 ± 7.2% (KO, P1) (n = 3, p = 0.17). Statistical analysis was done by two-sided Student's t-test. n.s.: no significance.
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f2: Nestin expression is normal in the KO mice.(a) Nestin staining in the cerebellum (E15.5) and cerebellar lobules (P1). Arrowheads indicate abnormal EGL thickness and folia formation (n = 7 pairs for each age). Scale bars: 50 μm. (b) Total nestin expression levels were measured by western blotting using GAPDH as the internal control. (c) Quantitation of nestin immunoreactivity. The nestin/GAPDH ratios were 61.7 ± 8.0% (WT, E15.5) and 60.6 ± 6.9% (KO, E15.5) (n = 3, p = 0.34); 62.6 ± 8.0% (WT, P1) and 64.1 ± 7.2% (KO, P1) (n = 3, p = 0.17). Statistical analysis was done by two-sided Student's t-test. n.s.: no significance.

Mentions: Since nestin is a well-known marker of neural stem cells (NSCs), and is expressed in undifferentiated CNS cells during development23, we investigated its expression in the cerebellum (E15.5 and P1) using immunocytochemical staining and western blotting. Confocal microscopy showed that nestin immunostaining in the KO cerebellum was almost identical to that in the WT cerebellum (Fig. 2a). When pooled cerebellar extracts were used to measure the total expression of the nestin protein (Fig. 2b), consistently, western blotting showed no difference in the nestin expression between WT and KO mice (Fig. 2c).


Essential roles of leucine-rich glioma inactivated 1 in the development of embryonic and postnatal cerebellum.

Xie YJ, Zhou L, Jiang N, Zhang N, Zou N, Zhou L, Wang Y, Cowell JK, Shen Y - Sci Rep (2015)

Nestin expression is normal in the KO mice.(a) Nestin staining in the cerebellum (E15.5) and cerebellar lobules (P1). Arrowheads indicate abnormal EGL thickness and folia formation (n = 7 pairs for each age). Scale bars: 50 μm. (b) Total nestin expression levels were measured by western blotting using GAPDH as the internal control. (c) Quantitation of nestin immunoreactivity. The nestin/GAPDH ratios were 61.7 ± 8.0% (WT, E15.5) and 60.6 ± 6.9% (KO, E15.5) (n = 3, p = 0.34); 62.6 ± 8.0% (WT, P1) and 64.1 ± 7.2% (KO, P1) (n = 3, p = 0.17). Statistical analysis was done by two-sided Student's t-test. n.s.: no significance.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4296302&req=5

f2: Nestin expression is normal in the KO mice.(a) Nestin staining in the cerebellum (E15.5) and cerebellar lobules (P1). Arrowheads indicate abnormal EGL thickness and folia formation (n = 7 pairs for each age). Scale bars: 50 μm. (b) Total nestin expression levels were measured by western blotting using GAPDH as the internal control. (c) Quantitation of nestin immunoreactivity. The nestin/GAPDH ratios were 61.7 ± 8.0% (WT, E15.5) and 60.6 ± 6.9% (KO, E15.5) (n = 3, p = 0.34); 62.6 ± 8.0% (WT, P1) and 64.1 ± 7.2% (KO, P1) (n = 3, p = 0.17). Statistical analysis was done by two-sided Student's t-test. n.s.: no significance.
Mentions: Since nestin is a well-known marker of neural stem cells (NSCs), and is expressed in undifferentiated CNS cells during development23, we investigated its expression in the cerebellum (E15.5 and P1) using immunocytochemical staining and western blotting. Confocal microscopy showed that nestin immunostaining in the KO cerebellum was almost identical to that in the WT cerebellum (Fig. 2a). When pooled cerebellar extracts were used to measure the total expression of the nestin protein (Fig. 2b), consistently, western blotting showed no difference in the nestin expression between WT and KO mice (Fig. 2c).

Bottom Line: The function of LGI1 in CNS development remains undefined.BrdU staining showed an inhibited proliferation of CGPs in KO embryos, which might be explained by the reduced Sonic hedgehog in embryos.Together, our results demonstrate that LGI1 is an essential player in the cerebellar development.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.

ABSTRACT
Leucine-rich glioma inactivated 1 (LGI1) is a secreted protein that interacts with ADAM transmembrane proteins, and its mutations are linked to human epilepsy. The function of LGI1 in CNS development remains undefined. Here, we report novel functions of LGI1 in the generation of cerebellar granule precursors (CGPs) and differentiation of radial glial cells (RGCs) in the cerebellum. A reduction in external granule layer thickness and defects in foliation were seen in embryonic and new-born LGI1 knockout (KO) mice. BrdU staining showed an inhibited proliferation of CGPs in KO embryos, which might be explained by the reduced Sonic hedgehog in embryos. In addition, the differentiation of RGCs into Bergmann glias was suppressed in KO mice. Enhanced Jagged1-Notch1 signaling in KO mice via reduced β-secretase proteolysis suggests that altered phenotype of RGCs is due to abnormal Notch1 signaling. Together, our results demonstrate that LGI1 is an essential player in the cerebellar development.

No MeSH data available.


Related in: MedlinePlus