Name: Gene-gun Transfection of Hippocampal Neurons (Video) | JoVE Journal
Uploaded: 2006-11-30T00:00:00.000+00:00
Duration: 5 min 44 s
Description: Learn gene-gun transfection of hippocampal neurons for neuroscience research using precise brain dissection techniques. This method enables efficient genetic manipulation of isolated hippocampal tissue.

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5 minutes

The authors have nothing to disclose.

gene-gun transfection of hippocampal neurons

Gene-gun Transfection of Hippocampal Neurons

Hi, I'm Kma. I work in Case Lab at Brown University Clean Slicer stage with 70%ethanol. Place plate insert into six.Well plate with medium and incubate at 37 degree whole brain with tweezers between midbrain and four brainin. Isolate hippocampus on one side, isolate hippocampus from the second side. Place hippocampi onto A PBS drop.Aspirate out the PBS make sagittal sections using the tissue slicer. Repeat transfer slicers onto a perish by squatting with PBS wash sections by adding and aspirating PBS and medium gently. Separate sections with tweezers.Wash sections with PBS and media. Again, transfer sections onto the plate and insert. Aspirate out the media in sections using tweezers.Attach gene gun hose to the helium cylinder. Attach barrel to gene gun. Fill the magazine with bullets.Insert magazine into the Jing gun. Adjust the pressure on the helium tank shoot. EGFP coated beads onto the sections.And now.

gene-gun-transfection-of-hippocampal-neurons

Research

JoVE Journal

Biology

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Preparation of Gene Gun Bullets and Biolistic Transfection of Neurons in Slice Culture

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Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons

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Nucleofection and Primary Culture of Embryonic Mouse Hippocampal and Cortical Neurons

Controlling the expansion of somatic stem cells is a major factor hampering their study and use in therapy. Here we describe a system to temporally control neural stem cells expansion during development and adulthood, which can be used to increase the number of neurons generated in the mouse brain.

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

Source: Keener, D. G., et al. Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons. J. Vis. Exp. (2020).This video demonstrates the process of single-cell electroporation to introduce plasmid DNA into neurons within mouse hippocampal slice cultures. The procedure involves precise pipette positioning, pressure adjustments, and electroporation pulses to transiently permeabilize the neuronal membrane,...

Gene Expression via Single-Cell Electroporation in Mouse Hippocampal Slice Cultures

Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this method on hard-to-transfect cells like primary neurons. Here we describe our detailed protocol for calcium phosphate transfection of hippocampal neurons cocultured with astroglial cells.

Calcium Phosphate Transfection of Primary Hippocampal Neurons

Recent improvements in organotypic brain slice preparations have permitted their exploitation for biotechnological applications. Organotypic slices maintain local structural characteristics of in vivo biology, including functional synaptic connections. Here we present a regioselective biolistic delivery method to label and genetically manipulate these slices.

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun

Here we present a protocol, designed to use chemogenetic tools to manipulate the activity of cortical interneuron progenitors transplanted into the cortex of early postnatal mice.

Transplantation of Chemogenetically Engineered Cortical Interneuron Progenitors into Early Postnatal Mouse Brains

Presented here is a protocol for single-cell electroporation that can deliver genes in both excitatory and inhibitory neurons across a range of in vitro hippocampal slice culture ages. Our approach provides precise and efficient expression of genes in individual cells, which can be used to examine cell-autonomous and intercellular functions.

Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons

This technique demonstrates an efficient way to prepare replication-defective retroviral stocks encoding a human oncogene, and subsequently used for induction of myeloproliferative disease in the mouse model.

Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells

Learn gene-gun transfection of hippocampal neurons for neuroscience research using precise brain dissection techniques. This method enables efficient genetic manipulation of isolated hippocampal tissue.

Gene-gun Transfection of Hippocampal Neurons

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