Time lapse recording of cortical interneuron migration in mouse organotypic brain slices and explants

Lepiemme, Fanny; Silva, Carla G.; Nguyen, Laurent

doi:10.1016/j.xpro.2021.100467

Cited by 6 publications

(4 citation statements)

References 7 publications

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“…The cell movement was tracked using ImageJ software and the Manual Tracking Plugin, and the movement parameters were calculated and analyzed in RStudio. The selected parameters are standard dynamics measurements previously shown in vivo ( 83 , 84 ) and in vitro ( 10 ).…”

Section: Methodsmentioning

confidence: 99%

Non–cell-autonomous regulation of interneuron specification mediated by extracellular vesicles

Pipicelli,

Baumann,

Di Giaimo

et al. 2023

Sci. Adv.

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Disruption in neurogenesis and neuronal migration can influence the assembly of cortical circuits, affecting the excitatory-inhibitory balance and resulting in neurodevelopmental and neuropsychiatric disorders. Using ventral cerebral organoids and dorsoventral cerebral assembloids with mutations in the extracellular matrix gene LGALS3BP , we show that extracellular vesicles released into the extracellular environment regulate the molecular differentiation of neurons, resulting in alterations in migratory dynamics. To investigate how extracellular vesicles affect neuronal specification and migration dynamics, we collected extracellular vesicles from ventral cerebral organoids carrying a mutation in LGALS3BP , previously identified in individuals with cortical malformations and neuropsychiatric disorders. These results revealed differences in protein composition and changes in dorsoventral patterning. Proteins associated with cell fate decision, neuronal migration, and extracellular matrix composition were altered in mutant extracellular vesicles. Moreover, we show that treatment with extracellular vesicles changes the transcriptomic profile in neural progenitor cells. Our results indicate that neuronal molecular differentiation can be influenced by extracellular vesicles.

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Section: Methodsmentioning

confidence: 99%

Non–cell-autonomous regulation of interneuron specification mediated by extracellular vesicles

Pipicelli,

Baumann,

Di Giaimo

et al. 2023

Sci. Adv.

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“…Ganglionic eminence explants were cultured on top of wild type cortical feeders as detailed in 83 . For this, a layer of wild type cortical neurons from E13.5 embryos were seeded on a glass bottom dish (MatTek or CELLView) previously coated sequentially with poly-L-Lysin (0.1mg/ml, Sigma-Aldrich) and laminin (10µg/ml, Sigma-Aldrich).…”

Section: Ganglionic Eminences Explant Culturementioning

confidence: 99%

“…For actin dynamic visualization, we used the pCAGGGS-lifeAct-Ruby plasmid (1µg/µl). Plasmids were prepared using a NucleoBond Xtra Maxi kit (Macherey-Nagel) and eletroporated ex-vivo as described by 83 . E13.5 embryos were decapitated and ganglionic eminences exposed.…”

Section: Ex Vivo Electroporationmentioning

confidence: 99%

Interneuron migration defects during corticogenesis contribute toDyrk1ahaploinsufficiency syndrome pathogenesis via actomyosin dynamics deregulations

Hinckelmann,

Dubos,

Artot

et al. 2023

Preprint

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Interneuron development is a crucial step of brain corticogenesis. When affected it often leads to brain dysfunctions, such as epilepsy, intellectual disabilities and autism spectrum disorder. Such defects are observed in theDYRK1A-haploinsufficiency syndrome, caused by mutations ofDYRK1A, and commonly associated to cortical excitatory/inhibitory imbalance. However, how this imbalance is established in this syndrome remains elusive. Here, using mouse models and live imaging, we show thatDyrk1aspecifically regulates the development of the cortical GABAergic system. Unlike projection excitatory neurons, we demonstrate that interneuron tangential migration relies on Dyrk1a dosage and kinase activity through a mechanism involving actomyosin cytoskeleton remodeling. Interestingly, we further demonstrate that mice with heterozygous inactivation ofDyrk1ain interneurons show behavioral defects and epileptic activity, recapitulating phenotypes observed in human patients. Altogether, these data highlight the critical role ofDyrk1ain the development of the GABAergic system and the pathophysiology ofDYRK1A-haploinsufficiency syndrome.

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“…We have employed this sparse labeling technique with brain slice methodologies [ 97 , 98 , 99 ] to conduct live imaging analyses of neural stem cell behavior [ 19 , 100 , 101 ]. Live imaging of the neural cells in brain slices after IUE has also been utilized in various other contexts, such as the analysis of neuronal migration [ 9 , 102 ], and extended to the subcellular level, such as the study of microtubule dynamics using EB3 [ 103 ], the Golgi apparatus [ 104 ], and visualization of the apical junctional components and centrosomes [ 59 ]. These advanced imaging techniques provide invaluable insights into human genetic disorders like lissencephaly, furthering our understanding of their underlying mechanisms.…”

Section: Spatiotemporal Expression Control and Lineage Tracing By Iuementioning

confidence: 99%

Advanced Techniques Using In Vivo Electroporation to Study the Molecular Mechanisms of Cerebral Development Disorders

Yang,

Shitamukai,

Yang

et al. 2023

IJMS

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The mammalian cerebral cortex undergoes a strictly regulated developmental process. Detailed in situ visualizations, imaging of these dynamic processes, and in vivo functional gene studies significantly enhance our understanding of brain development and related disorders. This review introduces basic techniques and recent advancements in in vivo electroporation for investigating the molecular mechanisms underlying cerebral diseases. In utero electroporation (IUE) is extensively used to visualize and modify these processes, including the forced expression of pathological mutants in human diseases; thus, this method can be used to establish animal disease models. The advent of advanced techniques, such as genome editing, including de novo knockout, knock-in, epigenetic editing, and spatiotemporal gene regulation, has further expanded our list of investigative tools. These tools include the iON expression switch for the precise control of timing and copy numbers of exogenous genes and TEMPO for investigating the temporal effects of genes. We also introduce the iGONAD method, an improved genome editing via oviductal nucleic acid delivery approach, as a novel genome-editing technique that has accelerated brain development exploration. These advanced in vivo electroporation methods are expected to provide valuable insights into pathological conditions associated with human brain disorders.

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Time lapse recording of cortical interneuron migration in mouse organotypic brain slices and explants

Cited by 6 publications

References 7 publications

Non–cell-autonomous regulation of interneuron specification mediated by extracellular vesicles

Non–cell-autonomous regulation of interneuron specification mediated by extracellular vesicles

Interneuron migration defects during corticogenesis contribute toDyrk1ahaploinsufficiency syndrome pathogenesis via actomyosin dynamics deregulations

Advanced Techniques Using In Vivo Electroporation to Study the Molecular Mechanisms of Cerebral Development Disorders

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