Myelination of rodent hippocampal neurons in culture

Gardner, Asa; Jukkola, Peter; Gu, Chen

doi:10.1038/nprot.2012.100

Cited by 34 publications

(34 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, dispersed mixed glial cultures established with neurons and oligodendrocytes from cortical tissues 4 or spinal cords 5,6 also contain similar contaminating cell types that are seen with slice cultures and that yield inconsistent amounts of myelin from culture to culture. In contrast, purified neuron-oligodendroglial coculture systems can be established using dorsal root ganglion neurons 7 , retinal ganglion neurons 8 or hippocampal neurons 9,10 . However, preparing these coculture systems involves tedious purification procedures and the maintenance of primary neurons.…”

Section: Introductionmentioning

confidence: 99%

A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers

et al. 2013

View full text Add to dashboard Cite

Current methods for studying oligodendrocyte myelination using primary neurons are limited by the time, cost and reproducibility of myelination in vitro. Nanofibers with diameters of >0.4 μm fabricated from electrospinning of liquid polystyrene are suitable scaffolds for concentric membrane wrapping by oligodendrocytes. With the advent of aligned electrospinning technology, nanofibers can be rapidly fabricated, standardized, and configured into various densities and patterns as desired. Notably, the minimally permissive culture environment of fibers provides investigators with an opportunity to explore the autonomous oligodendrocyte cellular processes underlying differentiation and myelination. The simplicity of the system is conducive to monitoring oligodendrocyte proliferation, migration, differentiation and membrane wrapping in the absence of neuronal signals. Here we describe protocols for the fabrication and preparation of nanofibers aligned on glass coverslips for the study of membrane wrapping by rodent oligodendrocytes. The entire protocol can be completed within 2 weeks.

show abstract

Section: Introductionmentioning

confidence: 99%

A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers

et al. 2013

View full text Add to dashboard Cite

show abstract

“…As many neurons are myelinated in the brain, we next examined whether myelination had any effect on the formation of MPS using a previously developed neuron-glia coculture system (31). Myelin sheaths were identified using an antibody against myelin basic protein (MBP).…”

Section: The Mps Structure Is Present In Multiple Specific Subtypes Omentioning

confidence: 99%

Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

Zhou

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

146

110

View full text Add to dashboard Cite

Actin, spectrin, and associated molecules form a periodic, submembrane cytoskeleton in the axons of neurons. For a better understanding of this membrane-associated periodic skeleton (MPS), it is important to address how prevalent this structure is in different neuronal types, different subcellular compartments, and across different animal species. Here, we investigated the organization of spectrin in a variety of neuronal-and glial-cell types. We observed the presence of MPS in all of the tested neuronal types cultured from mouse central and peripheral nervous systems, including excitatory and inhibitory neurons from several brain regions, as well as sensory and motor neurons. Quantitative analyses show that MPS is preferentially formed in axons in all neuronal types tested here: Spectrin shows a long-range, periodic distribution throughout all axons but appears periodic only in a small fraction of dendrites, typically in the form of isolated patches in subregions of these dendrites. As in dendrites, we also observed patches of periodic spectrin structures in a small fraction of glial-cell processes in four types of glial cells cultured from rodent tissues. Interestingly, despite its strong presence in the axonal shaft, MPS is disrupted in most presynaptic boutons but is present in an appreciable fraction of dendritic spine necks, including some projecting from dendrites where such a periodic structure is not observed in the shaft. Finally, we found that spectrin is capable of adopting a similar periodic organization in neurons of a variety of animal species, including Caenorhabditis elegans, Drosophila, Gallus gallus, Mus musculus, and Homo sapiens.ctin is critically involved in the regulation of neuronal polarization, differentiation, and growth of neuronal processes, cargo trafficking, and plasticity of synapses (1-3). Spectrin is an actin-binding protein that is important for the development and stabilization of axons and maintenance of neuronal polarization (4-6). In Caenorhabditis elegans, spectrin is important for the stability and integrity of axons under mechanical stress (4, 6) and for mechanosensation (6), and spectrin depletion results in axon breakage during animal locomotion (4). In Drosophila, spectrin has been shown to be involved in axonal path finding (7) and stabilization of presynaptic terminals (8). In mice, spectrin null mutations are embryonically lethal, and neurons with spectrin knockdown display defects in axonal initial segment assembly (5, 9, 10).Actin and spectrin form a 2D polygonal lattice structure underneath the membrane of erythrocytes (11). Recently, a novel form of actin-spectrin-based submembrane skeleton structure was discovered in neuronal axons (12) using superresolution STORM imaging (13,14). This membrane-associated periodic skeleton (MPS) has been observed in both fixed and live cultured neurons (12, 15, 16) and in brain tissue sections (12). In this structure, short actin filaments are organized into repetitive, ring-like structures that wrap around the circumference o...

show abstract

“…There is variety evidence showing that cortical and hippocampal neurons reach neurochemical maturation during 14-21 DIV [29][30][31]. On the othr hand, Price et al [26] demonstrated the total number of dendrites and total dendritic length as indices of neuronal maturation in cultured neurons.…”

Section: Effect Of Nlgn1 Knockdown On Maturation Of Primary Cortical mentioning

confidence: 98%

Prenatal Exposure to Histone Deacetylase Inhibitors Affects Gene Expression of Autism-Related Molecules and Delays Neuronal Maturation

et al. 2016

View full text Add to dashboard Cite

Valproic acid (VPA) is a multi-target drug and an inhibitor of histone deacetylase (HDAC). We have previously demonstrated that prenatal exposure to VPA at embryonic day 12.5 (E12.5), but not at E14.5, causes autism-like behavioral abnormalities in male mouse offspring. We have also found that prenatal VPA exposure causes transient histone hyperacetylation in the embryonic brain, followed by decreased neuronal cell numbers in the prefrontal and somatosensory cortices after birth. In the present study, we examined whether prenatal HDAC inhibition affects neuronal maturation in primary mouse cortical neurons. Pregnant mice were injected intraperitoneally with VPA (500 mg/kg) and the more selective HDAC inhibitor trichostatin A (TSA; 500 µg/kg) at E12.5 or E14.5, and primary neuronal cultures were prepared from the cerebral cortices of their embryos. Prenatal exposure to VPA at E12.5, but not at E14.5, decreased total number, total length, and complexity of neuronal dendrites at 14 days in vitro (DIV). The effects of VPA weakened at 21 DIV. Exposure to TSA at E12.5, but not at E14.5, also delayed maturation of cortical neurons. In addition, real-time quantitative PCR revealed that the prenatal exposure to TSA decreased neuroligin-1 (Nlgn1), Shank2, and Shank3 mRNA levels and increased contactin-associated protein-like 2 mRNA level. The delay in neuronal maturation was also observed in Nlgn1-knockdown cells, which were transfected with Nlgn1 siRNA. These findings suggest that prenatal HDAC inhibition causes changes in gene expression of autism-related molecules linked to a delay of neuronal maturation.

show abstract

Myelination of rodent hippocampal neurons in culture

Cited by 34 publications

References 33 publications

A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers

A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers

Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

Prenatal Exposure to Histone Deacetylase Inhibitors Affects Gene Expression of Autism-Related Molecules and Delays Neuronal Maturation

Contact Info

Product

Resources

About