Name: cortex-, hippocampus-, thalamus-, hypothalamus-, lateral septal nucleus- and striatum-specific in utero electroporation in the c57bl/6 mouse
Uploaded: 2016-01-18T14:00:00
Description: Watch this Scientific Journal Video about Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse at JoVE.com

Technical supported by Melanie Pfeifer and Nikolai Schmarowski (Institute for Microscopic Anatomy and Neurobiology, University Medical Center Mainz).

Ethics Statement: The handling of the mice and the experimental procedures were conducted in accordance with European, national and institutional guidelines for animal care.
1. In Utero Electroporation
Note: In utero electroporation was performed as previously published12,14. Therefore, the method is only described briefly in the following (Figure 1).
<ol>
	<li>Preparations
	<ol>
		<li>Prepare Fast Green colored endoto...

<ol>
	<li>Fukuchi-Shimogori, T., Grove, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neocortex+patterning+by+the+secreted+signaling+molecule+FGF8.">Neocortex patterning by the secreted signaling molecule FGF8.</a> Science. 294 (5544), 1071-1074 (2001).</li><li>Saito, T., Nakatsuji, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+transfer+into+the+embryonic+mouse+brain+using+in+vivo+electroporation.">Efficient gene transfer into the embryonic mouse brain using in vivo electroporation.</a> Dev Biol. 240 (1), 237-246 (2001).</li><li>Tabata, H., Nakajima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+in+utero+gene+transfer+system+to+the+developing+mouse+brain+using+electroporation:+Visualization+of+neuronal+migration+in+the+developing+cortex.">Efficient in utero gene transfer system to the developing mouse brain using electroporation: Visualization of neuronal migration in the developing cortex.</a> Neuroscience. 103 (4), 865-872 (2001).</li><li>LoTurco, J., Manent, J. B., Sidiqi, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+and+improved+tools+for+in+utero+electroporation+studies+of+developing+cerebral+cortex.">New and improved tools for in utero electroporation studies of developing cerebral cortex.</a> Cereb Cortex. 19 (July), i120-i125 (1991).</li><li>Nakahira, E., Yuasa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+generation,+migration,+and+differentiation+in+the+mouse+hippocampal+primoridium+as+revealed+by+enhanced+green+fluorescent+protein+gene+transfer+by+means+of+in+utero+electroporation.">Neuronal generation, migration, and differentiation in the mouse hippocampal primoridium as revealed by enhanced green fluorescent protein gene transfer by means of in utero electroporation.</a> J Comp Neurol. 483 (3), 329-340 (2005).</li><li>Navarro-Quiroga, I., Chittajallu, R., Gallo, V., Haydar, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term,+selective+gene+expression+in+developing+and+adult+hippocampal+pyramidal+neurons+using+focal+in+utero+electroporation.">Long-term, selective gene expression in developing and adult hippocampal pyramidal neurons using focal in utero electroporation.</a> J Neurosci. 27 (19), 5007-5011 (2007).</li><li>Borrell, V., Yoshimura, Y., Callaway, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+gene+delivery+to+telencephalic+inhibitory+neurons+by+directional+in+utero+electroporation.">Targeted gene delivery to telencephalic inhibitory neurons by directional in utero electroporation.</a> J Neurosci Methods. 143 (2), 151-158 (2005).</li><li>Kolk, S. M., de Mooij-Malsen, A. J., Martens, G. J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+Molecular+Approach+of+in+utero+Electroporation+to+Functionally+Decipher+Endophenotypes+in+Neurodevelopmental+Disorders.">Spatiotemporal Molecular Approach of in utero Electroporation to Functionally Decipher Endophenotypes in Neurodevelopmental Disorders.</a> FNMOL. 4 (November), 37 (2011).</li><li>Tomita, K., Kubo, K. I., Ishii, K., Nakajima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disrupted-in-schizophrenia-1+(Disc1)+is+necessary+for+migration+of+the+pyramidal+neurons+during+mouse+hippocampal+development.">Disrupted-in-schizophrenia-1 (Disc1) is necessary for migration of the pyramidal neurons during mouse hippocampal development.</a> Hu Mol Gen. 20 (14), 2834-2845 (2011).</li><li>Niwa, M., Kamiya, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Knockdown+of+DISC1+by+In+Utero+Gene+Transfer+Disturbs+Postnatal+Dopaminergic+Maturation+in+the+Frontal+Cortex+and+Leads+to+Adult+Behavioral+Deficits.">Knockdown of DISC1 by In Utero Gene Transfer Disturbs Postnatal Dopaminergic Maturation in the Frontal Cortex and Leads to Adult Behavioral Deficits.</a> Neuron. 65 (4), 480-489 (2010).</li><li>Nakahira, E., Kagawa, T., Shimizu, T., Goulding, M. D., Ikenaka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+evidence+that+ventral+forebrain+cells+migrate+to+the+cortex+and+contribute+to+the+generation+of+cortical+myelinating+oligodendrocytes.">Direct evidence that ventral forebrain cells migrate to the cortex and contribute to the generation of cortical myelinating oligodendrocytes.</a> Dev Biol. 291 (1), 123-131 (2006).</li><li>Baumgart, J., Grebe, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=C57BL/6-specific+conditions+for+efficient+in+utero+electroporation+of+the+central+nervous+system.">C57BL/6-specific conditions for efficient in utero electroporation of the central nervous system.</a> J Neurosci Methods. 240, 116-124 (2015).</li><li>Saito, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+electroporation+in+the+embryonic+mouse+central+nervous+system.">In vivo electroporation in the embryonic mouse central nervous system.</a> Nat Protc. 1 (3), 1552-1558 (2006).</li><li>Matsui, A., Yoshida, A. C., Kubota, M., Ogawa, M., Shimogori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+in+utero+electroporation:+controlled+spatiotemporal+gene+transfection.">Mouse in utero electroporation: controlled spatiotemporal gene transfection.</a> JoVE. (54), (2011).</li><li>Guedel, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Inhalation anesthesia: a fundamental guide. , (1937).</li><li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+Animal+Perfusion+Fixation+for+Rodents.">Whole Animal Perfusion Fixation for Rodents.</a> JoVE. (65), e3564 (2012).</li><li>Mizutani, K., Saito, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progenitors+resume+generating+neurons+after+temporary+inhibition+of+neurogenesis+by+Notch+activation+in+the+mammalian+cerebral+cortex.">Progenitors resume generating neurons after temporary inhibition of neurogenesis by Notch activation in the mammalian cerebral cortex.</a> Development. 132 (6), 1295-1304 (2005).</li><li>Tabata, H., Nakajima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Labeling+embryonic+mouse+central+nervous+system+cells+by+in+utero+electroporation.">Labeling embryonic mouse central nervous system cells by in utero electroporation.</a> Dev, Growth &amp; Differ. 50 (6), 507-511 (2008).</li></ol>

This protocol describes in detail how to transfect the retrosplenial cortex, the motor cortex, the somatosensory cortex, the piriform cortex, the cornu ammonis 1-3, the dentate gyrus, the striatum, the lateral septal nucleus, the thalamus and the hypothalamus of C75BL/6 mice. With all the provided information this is the first protocol, which supplies all necessary information to easily recreate transfections of these cerebral regions in the C57BL/6 mouse. Previous publications are mostly focused only on a few specific r...

Since the first description in 2001 by three independent groups 1-3 in utero electroporation has become a widely used standard tool for analyzing gene expression in the rodent central nervous system. Compared to the generation of knockout mice, which is, despite continuously improving techniques, still time and money consuming, the in utero electroporation appeals due to its simplicity. So, in utero electroporation enables fast and efficient gain- and loss-of-function studies 4.
To transfect the cerebral regions, the solution containing the negatively charged plasmid is injected into a ventricle. During the electric pulse, the negatively charged DNA migrates towards the positive pole and therefore the transfected region can be selected simply by altering the position of the positive pole. It has frequently been shown that numerous regions of the central nervous system can be targeted 3,5-8. For instance, recent studies show specific transfections of the hippocampus, the piriform cortex or the striatum 9-11. However, the information about the appropriate positions are often only scarcely standardized and are not always easy to transfer to different mouse strains.
Transfection of certain embryonic stages is far from trivial. Many influencing factors must be taken into consideration when choosing the set-up for specific in utero electroporation. First, to optimally transfect the respective embryonic stages, knowledge about the appropriate voltages is needed. High voltages decrease the survival rate, whereas low voltages reduce the transfection efficiency 2,3,12. Also the size of the electrode paddle plays a crucial role, because the use of electrode paddles that are too large results in reduced specificity or can cause death due to affection of the heart rhythm 4,12,13. The applied voltage and the size and the position of the electrode paddle are the most important features to consider, but there are also further factors influencing the outcome of the electroporation, like the applied amount of DNA-solution.
We have developed a detailed protocol which enables fast and efficient transfection of various cerebral regions of the C57BL/6 mouse 12. In this protocol detailed information about the voltages to be used and the size of the electrode paddle for enhanced specificity is provided. Further, information about the ventricle to be filled along with recommendations for the amount of plasmid solution and the position of the electrode is supplied. The indication of the detailed position information in a map and the further visualization of these positions enables straightforward specific and efficient in utero electroporation of the retrosplenial cortex, the motor cortex, the somatosensory cortex, the piriform cortex, the cornu ammonis 1-3, the dentate gyrus, the striatum, the lateral septal nucleus, the thalamus and the hypothalamus.

<table border="0" cellpadding="0" cellspacing="0" width="898">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:328px;">EndoFree Plasmid Maxi Kit</td>
			<td style="width:215px;">QIAGEN</td>
			<td style="width:188px;">12362</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fast Green</td>
			<td>Roth</td>
			<td>0301.1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pCAGGS</td>
			<td>Addgene</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">borosilicate glass capillaries (0.8-0.9 mm diameter)</td>
			<td>Wold Precision Instrument Inc.</td>
			<td>1B100F-4</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane (Forene)</td>
			<td>Abbott</td>
			<td>PZN 4831850</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Carprofen (Rimadyl)</td>
			<td>Pfizer GmbH</td>
			<td>approval number: 400684.00.00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">eye ointment (Bepanthen Augen und Nasensalbe)</td>
			<td>Bayer&#160;</td>
			<td>PZN 01578681</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">0.9% benzyl alcohol 0.9% saline solution</td>
			<td style="width:215px;">Pharmacy 
			of the University Medical Center Mainz</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">gauze (ES-Kompressen)</td>
			<td>Hartmann</td>
			<td>407835</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sterile 5-0 Perma-Hand Silk Suture</td>
			<td>Ethicon Johnson &#38; Johnson</td>
			<td>K890H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ring forceps 1/ 1.5 mm</td>
			<td>Fine Science Tools</td>
			<td>11101-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ring forceps 4.8/ 6 mm</td>
			<td>Fine Science Tools</td>
			<td>11106-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ring forceps 2.2/ 3 mm</td>
			<td>Fine Science Tools</td>
			<td>11103-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adson Forceps-Serrated Straight 12 cm</td>
			<td>Fine Science Tools</td>
			<td>1106-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IrisScissors-Delicate Straight-Sharp/Blunt 10 cm</td>
			<td>Fine Science Tools</td>
			<td>14028-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mayo-Stille Scissors-Straight 15 cm</td>
			<td>Fine Science Tools</td>
			<td>14012-15</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #5 Forceps-Inox</td>
			<td>Fine Science Tools</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo NeedleHolder-with Lock-Tungsten Carbide 14 cm</td>
			<td>Fine Science Tools</td>
			<td>12565-14</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Elektroporator CUY21 SC&#160;</td>
			<td>Nepa Gene Co.</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FST 250 Hot Bead Sterilizer</td>
			<td>Fine Science Tools</td>
			<td>18000-45</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microgrinder EG-44</td>
			<td>Narishige</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">P-97 Micropette Puller</td>
			<td>Sutter Instrument Company</td>
			<td>P-97</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum electrodes 650P 0.5 mm</td>
			<td>Nepagene</td>
			<td>CUY650P0.5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum electrodes 650P 3 mm</td>
			<td>Nepagene</td>
			<td>CUY650P3</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum electrodes 650P 5 mm</td>
			<td>Nepagene</td>
			<td>CUY650P5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum electrodes 650P 10 mm</td>
			<td>Nepagene</td>
			<td>CUY650P10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthesia system</td>
			<td>Rothacher-Medical GmbH</td>
			<td>CV-30511-3 Vapor 19.3</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Heating plate</td>
			<td>Rothacher-Medical GmbH</td>
			<td>HP-1M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temperature Controller 220V AC</td>
			<td>Rothacher-Medical GmbH</td>
			<td>TCAT-2LV</td>
			<td></td>
		</tr>
	</tbody>
</table>

cortex-, hippocampus-, thalamus-, hypothalamus-, lateral septal nucleus- and striatum-specific in utero electroporation in the c57bl/6 mouse

In utero electroporation is a widely used technique for fast and efficient spatiotemporal manipulation of various genes in the rodent central nervous system. Overexpression of desired genes is just as possible as shRNA mediated loss-of-function studies. Therefore it offers a wide range of applications. The feasibility to target particular cells in a distinct area further increases the range of potential applications of this very useful method. For efficiently targeting specific regions knowledge about the subtleties, such as the embryonic stage, the voltage to apply and most importantly the position of the electrodes, is indispensable.
Here, we provide a detailed protocol that allows for specific and efficient in utero electroporation of several regions of the C57BL/6 mouse central nervous system. In particular it is shown how to transfect regions the develop into the retrosplenial cortex, the motor cortex, the somatosensory cortex, the piriform cortex, the cornu ammonis 1-3, the dentate gyrus, the striatum, the lateral septal nucleus, the thalamus and the hypothalamus. For this information about the appropriate embryonic stage, the appropriate voltage for the corresponding embryonic stage is provided. Most importantly an angle-map, which indicates the appropriate position of the positive pole, is depicted. This standardized protocol helps to facilitate efficient in utero electroporation, which might also lead to a reduced number of animals.

Figure 2, shows examples for the specific in utero electroporation of the regions developing into the retrosplenial cortex, the motor cortex, the somatosensory cortex, the piriform cortex, the cornu ammonis 1-3, the dentate gyrus, the striatum, the lateral septal nucleus, the thalamus and the hypothalamus. The results of the transfections are shown next to the recommended angle (Figure 2). For better visualization of the angles in vivo the position of the electrode (0.5...

Watch this Scientific Journal Video about Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse at JoVE.com

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

This protocol describes in detail how to specifically transfect different regions in the C57BL/6 central nervous system via in utero electroporation. Included in this protocol are detailed instructions for transfections of regions that develop into the cortex, hippocampus, thalamus, hypothalamus, lateral septal nucleus and striatum.

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

In utero electroporation is a widely used technique for fast and efficient spatiotemporal manipulation of various genes in the rodent central nervous ...

The overall goal of this in vivo cell manipulation is to specifically modify cells of certain regions in the central nervous system of the mouse-like cortex, hippocampus, thalamus, hypothalamus, lateral septal nucleus, and striatum. This method can help to answer key questions concerning the development and function of the brain, for instance, through the identification of different signaling cascades and the determination of the level of cell differentiation. The main advantage of this technique is that it allows for fast and efficient manipulation of various cell types in the central nervous system without the time-consuming generation of genetically modified mice.Generally, the demonstration of this method is helpful, because individuals new to this method will often struggle with finding the right electrode position, which is critical for specific electroporation. To begin this procedure, cut open the abdominal cavity of an anesthetized pregnant mouse with a scalpel. Prevent the opened abdominal cavity from drying by moistening it with 0.9%warm methyl alcohol saline solution.Then, carefully extract the uterine horns using a pair of ring forceps. Next, position an E14 embryo so that there are no visible vessels above the puncture site. To have a better visualization, use a 5-millimeter electrode to locate the angle and position of the desired area in the brain for transfection.For the in utero electroporation of cortical area, slowly inject 1.5 to two microliters of the DNA solution in the left lateral ventricle. And ensure that the insertion depth is one to two millimeters, measured from the uterine wall. Then, check for blue-green coloring with a sharp demarcation.For better visualization, use a 5-millimeter electrode to locate the angle and position of the desired location in the brain for transfection. After that, place the center of the three-millimeter platinum electrode paddles just anterior of the ear primordia, and make sure that they are not placed above the vessels or the placenta. Subsequently, apply voltage to begin transfection.For the in utero electroporation of hippocampal formation, inject two microliters of DNA solution in the right lateral ventricle of an E15 embryo. Then, place the negative pole of the three to five millimeter platinum electrode paddles just anterior of the ear primordium, and the positive pole at the middle of the ear primordium. Then, apply voltage to begin transfection.For the in utero electroporation of the lateral septal nucleus and striatum, inject one microliter of DNA solution in the right lateral ventricle of an E12 embryo. Then, place the 5-millimeter electrodes at the level of the ear primordia. Subsequently, apply voltage to begin transfection.For the in utero electroporation of the thalamus and hypothalamus, inject one to 1.5 microliters of DNA solution in the lateral ventricle of an E12 to E13 embryo. Then, wait for two to three minutes for the solution to diffuse into the third ventricle. Next, place the 5 or three-millimeter electrodes just posterior to the ear primordia.After that, apply voltage to begin transfection. This schematic shows the appropriate positions for the positive and negative poles for transfecting cortical areas. And this one, shows the positions of the positive and negative pole for transfecting the hippocampal formations.The positions of the positive and negative pole for transfecting the striatum and the lateral septal nucleus are shown here. And the appropriate positions of the electrodes for transfecting the thalamus and the hypothalamus are shown here. The specificity of transfection depends on the electrode size.Increasing the size of electrode paddle reduces the specificity of the transfection. This is especially obvious in younger embryonic stages, but also visible in late embryonic stages. Once mastered, this technique can be done in 15 minutes if it is performed properly.While attempting this procedure, it's important to handle the embryos and the mothers with great care. After watching this video, you should have a good understanding of how to transfect specific regions in the central nervous system of the Black 6 mouse.

Research

JoVE Journal

Neuroscience

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural ...

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance ...

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetic  modification of specific regions of the developing mammalian brain is a very  powerful experimental approach. However, generating novel mouse ...

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by ...

Inducing Cre-lox Recombination in Mouse Cerebral Cortex Through In Utero Electroporation

Inducing Cre-lox Recombination in Mouse Cerebral Cortex Through In Utero Electroporation

Cell-autonomous neuronal functions of genes can be revealed by causing loss or gain of function of a gene in a small and sparse population of neurons. To ...

Cochlear Implant Surgery and Electrically-evoked Auditory Brainstem Response Recordings in C57BL/6 Mice

Cochlear implants (CIs) are neuroprosthetic devices that can provide a sense of hearing to deaf people. However, a CI cannot restore all aspects of ...

In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus

In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus

Microglia, the only immune cells resident in the brain, actively participate in neural circuit maintenance by modifying synapses and neuronal ...

LFP Recording in Mouse Brain for Cognitive Research and Drug Analysis

Dual Extracellular Recordings in the Mouse Hippocampus and Prefrontal Cortex

Author Spotlight: Studying Drug Impacts on Brain Signals Using Dual LFP Recording Protocol in Mice

The technique of recording local field potentials (LFPs) is an electrophysiological method used to measure the electrical activity of localized neuronal ...

Adolescent Social Stress Paradigm in C57BL&#47;6 Mice for Behavioral Research

Social Defeat Stress Model for Adolescent C57BL&#47;6 Male and Female Mice

Author Spotlight: Understanding Adolescent Social Adversity Effects on Neurodevelopment in Mice

Social adversity in adolescence is prevalent and can negatively impact mental health trajectories. Modeling social stress in adolescent male and female ...