NeuroMag Transfection Reagent is the first dedicated Magnetofection™ transfection reagent for neurons from 1 DIV to 21 DIV. It has proven to be extremely efficient in transfecting a large variety of primary neurons such as cortical, hippocampal, dorsal root ganglion and motor neurons with all types of nucleic acids. Moreover, high transfection efficiency was also achieved in primary astrocytes, oligodendrocyte precursors or neural stem cells as well as other cell lines (C6, B65, PC12, N2A...). Due to its unique properties, NeuroMag allows following transfected neurons during several days.
Great efficiency, ideal for primary neurons
Efficient from 0 DIV to 21 DIV
Non toxic and completely biodegradable: high transfected neurons viability
Ready-to-use, straightforward and rapid
For all types of nucleic acids
"High transfection efficiency on primary dopaminergic neurons at 21 DIV." Underhill SM et al, Neuron. 2014
“Achieve up to 30% of transfection efficiency on primary cortical neurons." Wang R et al, Neurobiol Dis. 2014
200 µL: up to 65 transfections with 1µg of DNA
500 µL: up to 165 transfections with 1µg of DNA
1 mL: up to 330 transfections with 1µg of DNA
#KC30800: Starting Kit with Super Magnetic plate + 200µL of NeuroMag
#KC30896 : Starting Kit with plate 96 magnets + 200µl of NeuroMag
Shipping Conditions: Room temperature
This reagent needs to be used with a magnetic plate
Starting Kit: Magnetic Plate + NeuroMag 200 µL
Starting Kit: Plate 96 magnets + NeuroMag 200 µL
Transfection of all types of nucleic acids: DNA, shRNA, siRNA, mRNA, oligonucleotides
Figure 1: Rat primary hippocampal neurons efficiently transfected with NeuroMag. Representative image of rat primary hippocampal neurons transfected with 1 μg pVectOZ-GFP and 3.5 μL NeuroMag in 24-well plate. Photos were taken under fluorescence 48H after transfection
Figure 2: Mouse cortical neuron expressing GFP (3 weeks in culture, 2-3 days after after magnetofection). Image aquisition: Nikon Ti-E microscope equipped with the A1 laser scanning confocal system and a 60x Apo TIRF objectif lens. Results were kindly provided by Dr. C. Charrier (Charrier C. et al., 2012, Cell, Vol 149, Issue 4, 923-935).
Completion of neuronal migration is critical for brain development. Kif21b is a plus-end-directed kinesin motor protein that promotes intracellular transport and controls microtubule dynamics in neurons. Here we report a physiological function of Kif21b during radial migration of projection neurons in the mouse developing cortex. In vivo analysis in mouse and live imaging on cultured slices demonstrate that Kif21b regulates the radial glia-guided locomotion of newborn neurons independently of its motility on microtubules. We show that Kif21b directly binds and regulates the actin cytoskeleton both in vitro and in vivo in migratory neurons. We establish that Kif21b-mediated regulation of actin cytoskeleton dynamics influences branching and nucleokinesis during neuronal locomotion. Altogether, our results reveal atypical roles of Kif21b on the actin cytoskeleton during migration of cortical projection neurons.
Petrova V., Nat Commun . 2020 Nov 5;11(1):5614. doi: 10.1038/s41467-020-19436-y.
Adult mammalian central nervous system axons have intrinsically poor regenerative capacity, so axonal injury has permanent consequences. One approach to enhancing regeneration is to increase the axonal supply of growth molecules and organelles. We achieved this by expressing the adaptor molecule Protrudin which is normally found at low levels in non-regenerative neurons. Elevated Protrudin expression enabled robust central nervous system regeneration both in vitro in primary cortical neurons and in vivo in the injured adult optic nerve. Protrudin overexpression facilitated the accumulation of endoplasmic reticulum, integrins and Rab11 endosomes in the distal axon, whilst removing Protrudin's endoplasmic reticulum localization, kinesin-binding or phosphoinositide-binding properties abrogated the regenerative effects. These results demonstrate that Protrudin promotes regeneration by functioning as a scaffold to link axonal organelles, motors and membranes, establishing important roles for these cellular components in mediating regeneration in the adult central nervous system.
Mingardi J., Int J Mol Sci . 2023 Jan 13;24(2):1552.
Stress is a key risk factor in the onset of neuropsychiatric disorders. The study of the mechanisms underlying stress response is important to understand the etiopathogenetic mechanisms and identify new putative therapeutic targets. In this context, microRNAs (miRNAs) have emerged as key regulators of the complex patterns of gene/protein expression changes in the brain, where they have a crucial role in the regulation of neuroplasticity, neurogenesis, and neuronal differentiation. Among them, miR-135a-5p has been associated with stress response, synaptic plasticity, and the antidepressant effect in different brain areas. Here, we used acute unavoidable foot-shock stress (FS) and chronic mild stress (CMS) on male rats to study whether miR-135a-5p was involved in stress-induced changes in the prefrontal cortex (PFC). Both acute and chronic stress decreased miR-135a-5p levels in the PFC, although after CMS the reduction was induced only in animals vulnerable to CMS, according to a sucrose preference test. MiR-135a-5p downregulation in the primary neurons reduced dendritic spine density, while its overexpression exerted the opposite effect. Two bioinformatically predicted target genes, Kif5c and Cplx1/2, were increased in FS rats 24 h after stress. Altogether, we found that miR-135a-5p might play a role in stress response in PFC involving synaptic mechanisms.
KIF21B is a kinesin protein that promotes intracellular transport and controls microtubule dynamics. We report three missense variants and one duplication in KIF21B in individuals with neurodevelopmental disorders associated with brain malformations, including corpus callosum agenesis (ACC) and microcephaly. We demonstrate, in vivo, that the expression of KIF21B missense variants specifically recapitulates patients' neurodevelopmental abnormalities, including microcephaly and reduced intra- and inter-hemispheric connectivity. We establish that missense KIF21B variants impede neuronal migration through attenuation of kinesin autoinhibition leading to aberrant KIF21B motility activity. We also show that the ACC-related KIF21B variant independently perturbs axonal growth and ipsilateral axon branching through two distinct mechanisms, both leading to deregulation of canonical kinesin motor activity. The duplication introduces a premature termination codon leading to nonsense-mediated mRNA decay. Although we demonstrate that Kif21b haploinsufficiency leads to an impaired neuronal positioning, the duplication variant might not be pathogenic. Altogether, our data indicate that impaired KIF21B autoregulation and function play a critical role in the pathogenicity of human neurodevelopmental disorder.
Protein quality control pathways have evolved to ensure the fidelity of protein synthesis and efficiently clear potentially toxic protein species. Defects in ribosome-associated quality control and its associated factors have been implicated in the accumulation of aberrant proteins and neurodegeneration. C9orf72 repeat-associated non-AUG translation has been suggested to involve inefficient translation elongation, lead to ribosomal pausing and activation of ribosome-associated quality control pathways. However, the role of the ribosome-associated quality control complex in the processing of proteins generated through this non-canonical translation is not well understood. Here we use reporter constructs containing the C9orf72-associated hexanucleotide repeat, ribosome-associated quality control complex deficient cell models and stain for ribosome-associated quality control markers in C9orf72-expansion carrier human tissue to understand its role in dipeptide-repeat protein pathology. Our studies show that canonical ribosome-associated quality control substrates products are efficiently cleared by the ribosome-associated quality control complex in mammalian cells. Furthermore, using stalling reporter constructs, we show that repeats associated with the C9orf72-expansion induce ribosomal stalling when arginine (R)-rich dipeptide-repeat proteins are synthesized in a length-dependent manner. However, despite triggering this pathway, these arginine-rich dipeptide-repeat proteins are not efficiently processed by the core components of the ribosome-associated quality control complex (listerin, nuclear-export mediator factor and valosin containing protein) partly due to lack of lysine residues, which precludes ubiquitination. Deficient processing by this complex may be implicated in C9orf72-expansion associated disease as dipeptide-repeat protein inclusions were observed to be predominantly devoid of ubiquitin and co-localize with nuclear-export mediator factor in mutation carriers' frontal cortex and cerebellum tissue. These findings suggest that impaired processing of these arginine-rich dipeptide-repeat proteins derived from repeat-associated non-AUG translation by the ribosome-associated quality control complex may contribute to protein homeostasis dysregulation observed in C9orf72-expansion amyotrophic lateral sclerosis and frontotemporal degeneration neuropathogenesis.