scholarly journals CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using In Utero Electroporation-based Gene Transfer

10.3791/57311 ◽  
2018 ◽  
Author(s):  
Weijun Feng ◽  
Lena Herbst ◽  
Peter Lichter ◽  
Stefan M. Pfister ◽  
Hai-Kun Liu ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Granule Cells ◽  
Cerebellar Granule Cells ◽  
Function Analysis ◽  
In Utero ◽  
Loss Of Function ◽  
Cerebellar Granule

scholarly journals Deletion of Mea6 in Cerebellar Granule Cells Impairs Synaptic Development and Motor Performance

2021 ◽  
Vol 8 ◽  
Author(s):  
Xin-Tai Wang ◽  
Lin Zhou ◽  
Xin-Yu Cai ◽  
Fang-Xiao Xu ◽  
Zhi-Heng Xu ◽  
...  
Keyword(s):  
Purkinje Cell ◽  
Motor Performance ◽  
Intracellular Transport ◽  
Granule Cells ◽  
Cerebellar Granule Cells ◽  
Glutamate Transporter ◽  
Loss Of Function ◽  
Cerebellar Granule ◽  
Fahr’S Disease ◽  
Fahr's Disease

The cerebellum is conceptualized as a processor of complex movements. Many diseases with gene-targeted mutations, including Fahr’s disease associated with the loss-of-function mutation of meningioma expressed antigen 6 (Mea6), exhibit cerebellar malformations, and abnormal motor behaviors. We previously reported that the defects in cerebellar development and motor performance of Nestin-Cre;Mea6F/Fmice are severer than those of Purkinje cell-targeted pCP2-Cre;Mea6F/Fmice, suggesting that Mea6 acts on other types of cerebellar cells. Hence, we investigated the function of Mea6 in cerebellar granule cells. We found that mutant mice with the specific deletion ofMea6in granule cells displayed abnormal posture, balance, and motor learning, as indicated in footprint, head inclination, balanced beam, and rotarod tests. We further showed that Math1-Cre;Mea6F/Fmice exhibited disrupted migration of granule cell progenitors and damaged parallel fiber-Purkinje cell synapses, which may be related to impaired intracellular transport of vesicular glutamate transporter 1 and brain-derived neurotrophic factor. The present findings extend our previous work and may help to better understand the pathogenesis of Fahr’s disease.

scholarly journals Single-cell spatial transcriptomic analysis reveals common and divergent features of developing postnatal granule cerebellar cells and medulloblastoma

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Wenqin Luo ◽  
Guan Ning Lin ◽  
Weichen Song ◽  
Yu Zhang ◽  
Huadong Lai ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Granule Cells ◽  
Cerebellar Granule Cells ◽  
Granule Neurons ◽  
Cerebellar Granule ◽  
Granule Neuron ◽  
Proliferation And Differentiation ◽  
Single Cell Rna Sequencing ◽  
Cerebellar Cells

Abstract Background Cerebellar neurogenesis involves the generation of large numbers of cerebellar granule neurons (GNs) throughout development of the cerebellum, a process that involves tight regulation of proliferation and differentiation of granule neuron progenitors (GNPs). A number of transcriptional regulators, including Math1, and the signaling molecules Wnt and Shh have been shown to have important roles in GNP proliferation and differentiation, and deregulation of granule cell development has been reported to be associated with the pathogenesis of medulloblastoma. While the progenitor/differentiation states of cerebellar granule cells have been broadly investigated, a more detailed association between developmental differentiation programs and spatial gene expression patterns, and how these lead to differential generation of distinct types of medulloblastoma remains poorly understood. Here, we provide a comparative single-cell spatial transcriptomics analysis to better understand the similarities and differences between developing granule and medulloblastoma cells. Results To acquire an enhanced understanding of the precise cellular states of developing cerebellar granule cells, we performed single-cell RNA sequencing of 24,919 murine cerebellar cells from granule neuron-specific reporter mice (Math1-GFP; Dcx-DsRed mice). Our single-cell analysis revealed that there are four major states of developing cerebellar granule cells, including two subsets of granule progenitors and two subsets of differentiating/differentiated granule neurons. Further spatial transcriptomics technology enabled visualization of their spatial locations in cerebellum. In addition, we performed single-cell RNA sequencing of 18,372 cells from Patched+/− mutant mice and found that the transformed granule cells in medulloblastoma closely resembled developing granule neurons of varying differentiation states. However, transformed granule neuron progenitors in medulloblastoma exhibit noticeably less tendency to differentiate compared with cells in normal development. Conclusion In sum, our study revealed the cellular and spatial organization of the detailed states of cerebellar granule cells and provided direct evidence for the similarities and discrepancies between normal cerebellar development and tumorigenesis.

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