scholarly journals Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

10.3791/51070 ◽  
2014 ◽  
Author(s):  
Anna Holubowska ◽  
Chaitali Mukherjee ◽  
Mayur Vadhvani ◽  
Judith Stegmüller
Keyword(s):  
Genetic Manipulation ◽  
Neuronal Morphology ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Cerebellar Granule ◽  
And Migration

scholarly journals Neurabin-I Is Phosphorylated by Cdk5: Implications for Neuronal Morphogenesis and Cortical Migration

2007 ◽  
Vol 18 (11) ◽  
pp. 4327-4342 ◽  
Author(s):  
Frédéric Causeret ◽  
Tom Jacobs ◽  
Mami Terao ◽  
Owen Heath ◽  
Mikio Hoshino ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Pyramidal Neurons ◽  
Normal Development ◽  
Phosphorylation Site ◽  
Neuronal Morphology ◽  
Actin Binding ◽  
Neuronal Morphogenesis ◽  
And Migration

The correct morphology and migration of neurons, which is essential for the normal development of the nervous system, is enabled by the regulation of their cytoskeletal elements. We reveal that Neurabin-I, a neuronal-specific F-actin–binding protein, has an essential function in the developing forebrain. We show that gain and loss of Neurabin-I expression affect neuronal morphology, neurite outgrowth, and radial migration of differentiating cortical and hippocampal neurons, suggesting that tight regulation of Neurabin-I function is required for normal forebrain development. Importantly, loss of Neurabin-I prevents pyramidal neurons from migrating into the cerebral cortex, indicating its essential role during early stages of corticogenesis. We demonstrate that in neurons Rac1 activation is affected by the expression levels of Neurabin-I. Furthermore, the Cdk5 kinase, a key regulator of neuronal migration and morphology, directly phosphorylates Neurabin-I and controls its association with F-actin. Mutation of the Cdk5 phosphorylation site reduces the phenotypic consequences of Neurabin-I overexpression both in vitro and in vivo, suggesting that Neurabin-I function depends, at least in part, on its phosphorylation status. Together our findings provide new insight into the signaling pathways responsible for controlled changes of the F-actin cytoskeleton that are required for normal development of the forebrain.

scholarly journals Plexin-B2 controls the timing of differentiation and the motility of cerebellar granule neurons

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Eljo Van Battum ◽  
Celine Heitz-Marchaland ◽  
Yvrick Zagar ◽  
Stéphane Fouquet ◽  
Rohini Kuner ◽  
...  
Keyword(s):  
3D Imaging ◽  
Imaging Techniques ◽  
Molecular Layer ◽  
Conditional Knockout ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Cerebellar Granule ◽  
External Granule Layer ◽  
Purkinje Cell Dendrites

Plexin-B2 deletion leads to aberrant lamination of cerebellar granule neurons (CGNs) and Purkinje cells. Although in the cerebellum Plexin-B2 is only expressed by proliferating CGN precursors in the outer external granule layer (oEGL), its function in CGN development is still elusive. Here, we used 3D imaging, in vivo electroporation and live-imaging techniques to study CGN development in novel cerebellum-specific Plxnb2 conditional knockout mice. We show that proliferating CGNs in Plxnb2 mutants not only escape the oEGL and mix with newborn postmitotic CGNs. Furthermore, motility of mitotic precursors and early postmitotic CGNs is altered. Together, this leads to the formation of ectopic patches of CGNs at the cerebellar surface and an intermingling of normally time-stamped parallel fibers in the molecular layer (ML), and aberrant arborization of Purkinje cell dendrites. There results suggest that Plexin-B2 restricts CGN motility and might have a function in cytokinesis.

Actin disruption alters the localization of tau in the growth cones of cerebellar granule neurons

2000 ◽  
Vol 113 (15) ◽  
pp. 2797-2809
Author(s):  
J.F. Zmuda ◽  
R.J. Rivas
Keyword(s):  
Growth Cone ◽  
Growth Cones ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Filamentous Actin ◽  
Cerebellar Granule ◽  
Single Axon ◽  
Peripheral Domain ◽  
The Way

Cultured cerebellar granule neurons initially extend a single axon, followed by the extension of a second axon to attain a bipolar morphology. Differentiation culminates with the extension of several short dendrites from the cell body. In the present study, we determined the location of the dephosphorylated form of the microtubule-associated protein tau (dtau) within the growth cones of newly forming axons and examined whether this localization was influenced by the actin cytoskeleton. Following elongation of the initial axon at 2–3 days in vitro, dtau immunoreactivity was present along the entire length of the axon, becoming most intense just proximal to the growth cone. Dtau labeling dropped off dramatically along the microtubules of the growth cone and was undetectable along the most distal tips of these microtubules. As the initial axon continued to elongate at 3–4 days in vitro, the actin-rich growth cone peripheral domain characteristically underwent a dramatic reduction in size. Dtau immunoreactivity extended all the way to the most distal tips of the microtubules in the growth cones of these cells. Cytochalasin D and latrunculin A mimicked the effects of this characteristic reduction in growth cone size with regard to dtau localization in the growth cone. Depolymerization of filamentous actin caused the collapse of the peripheral domain and allowed dtau to bind all the way to the most distal tips of microtubules in the axon. Upon removal of the drugs, the peripheral domain of the growth cone rapidly re-formed and dtau was once again excluded from the most distal regions of growth cone microtubules. These findings suggest a novel role for actin in determining the localization of the microtubule-associated protein τ within the growth cones of neurons.

scholarly journals Reciprocal autoregulation by NFI occupancy and ETV1 promotes the developmental expression of dendrite-synapse genes in cerebellar granule neurons

2016 ◽  
Vol 27 (9) ◽  
pp. 1488-1499 ◽  
Author(s):  
Baojin Ding ◽  
John W. Cave ◽  
Paul R. Dobner ◽  
Debra Mullikin-Kilpatrick ◽  
Marina Bartzokis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Target Genes ◽  
Feedback Regulation ◽  
Developmental Expression ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Cerebellar Granule ◽  
Temporal Gene Expression

Nuclear Factor One (NFI) transcription factors regulate temporal gene expression required for dendritogenesis and synaptogenesis via delayed occupancy of target promoters in developing cerebellar granule neurons (CGNs). Mechanisms that promote NFI temporal occupancy have not been previously defined. We show here that the transcription factor ETV1 directly binds to and is required for expression and NFI occupancy of a cohort of NFI-dependent genes in CGNs maturing in vivo. Expression of ETV1 is low in early postnatal cerebellum and increases with maturation, mirroring NFI temporal occupancy of coregulated target genes. Precocious expression of ETV1 in mouse CGNs accelerated onset of expression and NFI temporal occupancy of late target genes and enhanced Map2(+) neurite outgrowth. ETV1 also activated expression and NFI occupancy of the Etv1 gene itself, and this autoregulatory loop preceded ETV1 binding and activation of other coregulated target genes in vivo. These findings suggest a potential model in which ETV1 activates NFI temporal binding to a subset of late-expressed genes in a stepwise manner by initial positive feedback regulation of the Etv1 gene itself followed by activation of downstream coregulated targets as ETV1 expression increases. Sequential transcription factor autoregulation and subsequent binding to downstream promoters may provide an intrinsic developmental timer for dendrite/synapse gene expression.

Chemotherapy-induced cell death in primary cerebellar granule neurons but not in astrocytes: in vitro paradigm of differential neurotoxicity

2004 ◽  
Vol 91 (5) ◽  
pp. 1067-1074 ◽  
Author(s):  
Antje Wick ◽  
Wolfgang Wick ◽  
Johannes Hirrlinger ◽  
Ellen Gerhardt ◽  
Ralf Dringen ◽  
...  
Keyword(s):  
Cell Death ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Cerebellar Granule
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