Primary Culture of Neurons Isolated from Embryonic Mouse Cerebellum

Shabanipour, Shahin; Dalvand, Azadeh; Jiao, Xun; Balaei, Maryam Rahimi; Chung, Siu-Leung; Kong, Jiming; Bigio, Marc R. Del; Marzban, Hassan

doi:10.3791/60168

Cited by 15 publications

(14 citation statements)

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“…Five distinct animal harvests were used to isolate primary cortical neurons. Two pregnant outbred C57BL/6 mice (E15‐16) were obtained from the Royan Institute's animal center and sacrificed via cervical dislocation for each harvest (Shabanipour et al, 2019; Xu et al, 2012). The uterus was immediately dissected, and six to eight embryos were blindly obtained for further isolation.…”

Section: Methodsmentioning

confidence: 99%

Tau nuclear translocation is a leading step in tau pathology process through P53 stabilization and nucleolar dispersion

Roqanian

Ahmadian

Nabavi

et al. 2022

J of Neuroscience Research

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Tau protein abnormalities are associated with various neurodegenerative disorders, including Alzheimer's disease (AD) and traumatic brain injury (TBI). In tau‐overexpressing SHSY5Y cells and iPSC‐derived neuron models of frontotemporal dementia (FTD), axonal tau translocates into the nuclear compartment, resulting in neuronal dysfunction. Despite extensive research, the mechanisms by which tau translocation results in neurodegeneration remain elusive thus far. We studied the nuclear displacement of different P‐tau species [Cis phosphorylated Thr231‐tau (cis P‐tau), phosphorylated Ser202/Thr205‐tau (AT8 P‐tau), and phosphorylated Thr212/Ser214‐tau (AT100 P‐tau)] at various time points using starvation in primary cortical neurons and single severe TBI (ssTBI) in male mouse cerebral cortices as tauopathy models. While all P‐tau species translocated into the somatodendritic compartment in response to stress, cis P‐tau did so more rapidly than the other species. Notably, nuclear localization of P‐tau was associated with p53 apoptotic stabilization and nucleolar stress, both of which resulted in neurodegeneration. In summary, our findings indicate that P‐tau nuclear translocation results in p53‐dependent apoptosis and nucleolar dispersion, which is consistent with neurodegeneration.

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Section: Methodsmentioning

confidence: 99%

Tau nuclear translocation is a leading step in tau pathology process through P53 stabilization and nucleolar dispersion

Roqanian

Ahmadian

Nabavi

et al. 2022

J of Neuroscience Research

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“…The widely used volatile anesthetic agent isoflurane (2%) was used for 5 min to sedate pregnant rats in a desiccant chamber. Once anesthetic depth was controlled by evaluating the foot reflex, rats were sacrificed by cervical dislocation, and the brains from embryos were obtained immediately after removal from their embryonic sacs (Shabanipour et al., 2019; Tomassoni‐Ardori et al, 2020). Experimental procedures were approved by the Bioethical and Biosafety Committee of the Faculty of Biological Sciences of the Pontificia Universidad Católica de Chile with the ethical approval 160720014.…”

Section: Methodsmentioning

confidence: 99%

Wnt5a modulates dendritic spine dynamics through the regulation of Cofilin via small Rho GTPase activity in hippocampal neurons

Vallejo

Lindsay

González-Billault

et al. 2021

Journal of Neurochemistry

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Dendritic spines are small, actin‐rich protrusions that act as the receiving sites of most excitatory inputs in the central nervous system. The remodeling of the synapse architecture is mediated by actin cytoskeleton dynamics, a process precisely regulated by the small Rho GTPase family. Wnt ligands exert their presynaptic and postsynaptic effects during formation and consolidation of the synaptic structure. Specifically, Wnt5a has been identified as an indispensable synaptogenic factor for the regulation and organization of the postsynaptic side; however, the molecular mechanisms through which Wnt5a induces morphological changes resulting from actin cytoskeleton dynamics within dendritic spines remain unclear. In this work, we employ primary rat hippocampal cultures and HT22 murine hippocampal neuronal cell models, molecular and pharmacological tools, and fluorescence microscopy (laser confocal and epifluorescence) to define the Wnt5a‐induced molecular signaling involved in postsynaptic remodeling mediated via the regulation of the small Rho GTPase family. We report that Wnt5a differentially regulates the phosphorylation of Cofilin in neurons through both Ras‐related C3 botulinum toxin substrate 1 and cell division cycle 42 depending on the subcellular compartment and the extracellular calcium levels. Additionally, we demonstrate that Wnt5a increases the density of dendritic spines and promotes their maturation via Ras‐related C3 botulinum toxin substrate 1. Accordingly, we find that Wnt5a requires the combined activation of small Rho GTPases to increase the levels of filamentous actin, thus promoting the stability of actin filaments. Altogether, these results provide evidence for a new mechanism by which Wnt5a may target actin dynamics, thereby regulating the subsequent morphological changes in dendritic spine architecture.

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“…To visualize the arrangement of PCs and BGCs, the cerebellar cortex was immunostained by anti-Calb1 and S100B, respectively, at P2, corresponding to the PC cluster stage, and P7, when PCs are dispersed and organized as a monolayer. In the mouse cerebellum, Calb1 is a specific marker of PCs (e.g., Baimbridge and Miller, 1982;Shabanipour et al, 2019), while S100B was used as a marker of both mature BGCs and their precursors (Raponi et al, 2007;Koirala and Corfas, 2010). At P2, a double immunofluorescence staining with anti-Calb1 and anti-S100B in both the wt (Figure 2A) and nax (Figure 2B) cerebellum shows that PCs form a multicellular cluster.…”

Section: Excessive Purkinje Cell Migration and Bergmann Glial Cell Positioning In The Nax Cerebellar Cortexmentioning

confidence: 99%

Upregulation of Neural Cell Adhesion Molecule 1 and Excessive Migration of Purkinje Cells in Cerebellar Cortex

Shabanipour

Jiao²,

Rahimi-Balaei³

et al. 2022

Front. Neurosci.

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Purkinje cells (PCs) are large GABAergic projection neurons of the cerebellar cortex, endowed with elaborate dendrites that receive a multitude of excitatory inputs. Being the only efferent neuron of the cerebellar cortex, PCs project to cerebellar nuclei and control behaviors ranging from movement to cognition and social interaction. Neural cell adhesion molecule 1 (NCAM1) is widely expressed in the embryonic and postnatal development of the brain and plays essential roles in neuronal migration, axon pathfinding and synapse assembly. However, despite its high expression levels in cerebellum, little is known to date regarding the role(s) of NCAM1 in PCs development. Among other aspects, elucidating how the expression of NCAM1 in PCs could impact their postnatal migration would be a significant achievement. We analyzed the Acp2 mutant mouse (nax: naked and ataxia), which displays excessive PC migration into the molecular layer, and investigated how the excessive migration of PCs along Bergmann glia could correlate to NCAM1 expression pattern in early postnatal days. Our Western blot and RT-qPCR analysis of the whole cerebellum show that the protein and mRNA of NCAM1 in wild type are not different during PC dispersal from the cluster stage to monolayer formation. However, RT-qPCR analysis from FACS-based isolated PCs shows that Ncam1 is significantly upregulated when PCs fail to align and instead overmigrate into the molecular layer. Our results suggest two alternative interpretations: (1) NCAM1 promotes excessive PC migration along Bergmann glia, or (2) NCAM1 upregulation is an attempt to prevent PCs from invading the molecular layer. If the latter scenario proves true, NCAM1 may play a key role in PC monolayer formation.

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Primary Culture of Neurons Isolated from Embryonic Mouse Cerebellum

Cited by 15 publications

References 0 publications

Tau nuclear translocation is a leading step in tau pathology process through P53 stabilization and nucleolar dispersion

Tau nuclear translocation is a leading step in tau pathology process through P53 stabilization and nucleolar dispersion

Wnt5a modulates dendritic spine dynamics through the regulation of Cofilin via small Rho GTPase activity in hippocampal neurons

Upregulation of Neural Cell Adhesion Molecule 1 and Excessive Migration of Purkinje Cells in Cerebellar Cortex

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