Neural Tube Transfection
Neural Tube Transfection - Related Videos
11:24
Transfecting Human Neural Stem Cells with the Amaxa Nucleofector
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Cited by 10 •
2007
Introducing a gene of interest into a cell is a powerful method for elucidating its function in vivo. This protocol describes an efficient method of transfecting a culture of human neural stem/precursor cells (hNSPCs) using the Nucleofector electroporation apparatus made by Amaxa.
12:48
Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube
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Cited by 26 •
2012
Isolation of embryonic neural crest from the neural tube facilitates the use of in vitro methods for studying migration, self-renewal, and multipotency of neural crest.
07:37
Neural Tube Closure in Mouse Whole Embryo Culture
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Cited by 22 •
2011
A method allowing for direct pharmacological manipulation of mouse embryos during neurulation that bypasses maternal metabolism is described. The technique can be adapted to study different aspects of neurulation by varying the time point and pharmacological agent.
09:57
Dual Labeling of Neural Crest Cells and Blood Vessels Within Chicken Embryos Using ChickGFP Neural Tube Grafting and Carbocyanine Dye DiI Injection
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Cited by 11 •
2015
Here we report dual labeling of neural crest cells and blood vessels using chickGFP neural tube intraspecies grafting combined with intra-vascular DiI injection. This experimental technique allows us to simultaneously visualize and study development of the NCC-derived (enteric) nervous system and the vascular system, during organogenesis.
07:28
An Introduction to Transfection
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2023
Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the...
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Brief electrical pulses transiently permeabilize the membranes of cells near the targeted side of the neural tube. This temporary change allows plasmid DNA placed in the lumen to enter those cells without permanently disrupting membrane integrity. Because uptake depends on the relationship between DNA placement and pulse application, the approach provides spatial control over which developing neural tissue receives the introduced genetic material.
Side-specific targeting creates a spatial comparison within the developing tissue. Cells exposed to the plasmid can be evaluated alongside neighboring or corresponding cells that were not targeted, helping researchers associate altered protein expression or developmental behavior with the introduced gene. This arrangement is particularly useful for examining localized effects on cell fate, migration, differentiation, axon growth, or circuit assembly.
The plasmid can be selected to alter protein expression, label particular cell populations, or regulate developmental pathways. These different uses allow the same transfection strategy to support either visualization or functional investigation. By changing the introduced genetic material, researchers can connect gene activity with the behavior and development of neural cells in their native, organized tissue context.
The method introduces genetic material at a chosen stage of neural development, so its effects can be examined in relation to ongoing formation of the nervous system. This timing helps researchers study processes such as differentiation, migration, axon growth, and circuit assembly as they occur. Temporal control therefore complements spatial targeting, linking when and where gene activity influences developing neural tissue.
A common workflow places plasmid DNA into the neural tube lumen and then applies brief electrical pulses to the selected side. The pulses transiently permeabilize nearby cell membranes, enabling DNA uptake. Researchers subsequently examine the targeted tissue for changes in protein expression, labeled cell populations, or developmental outcomes, depending on the purpose of the introduced gene.
Researchers would choose neural tube transfection when they need access to developing neural tissue with both spatial and temporal control. It can support studies of neural cell fate, migration, differentiation, axon growth, and circuit assembly. The technique is especially informative when a gene must be introduced into a defined region so its contribution to nervous system formation or function can be evaluated.