The authors would like to thank John Marston for expert animal husbandry and Dr. Marco A. Passini for technical assistance at the early stages of this project. This work was funded by grant from the National Institutes of Health to C.L.L. (R01NS41584; R01HD054413).

1. Intracerebroventricular injection
<ol>
<li>The first step is the preparation of the injection stock solutions; these solutions are viral vector, plasmid DNA, drugs, and should be injected under sterile conditions. </li> 
<li>Mix a desired titer of viral vector (5-7 &mu;L total) with 0.05% w/v trypan blue in PBS for visualization of the injection site.</li>
<li>Plasmid DNA solution (5 l &mu;L total) contains D-(+)-glucose 20% (w/v) (1 &mu;L), trypan blue (0.05%) PBS (1 &mu;L), plasmid (&tilde;5 &tilde;g/&tilde;L) (2 ...

<ol>
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Research using mouse models of disease often requires the administration of drugs or other substances to neonates. In this video, we demonstrate the step-by step procedures involving two types of injection strategies that can be used to target the CNS: 1) direct injection into the CNS utilizing intracerebroventricular (ICV) injection; or 2) IV injection targeting the temporal/facial vein. The timing of these injections is of great importance. Since the ICV injections are freehand, the skull must be relatively malleable. ...

<table border="1">
	<tbody>
		<tr>
			<th>Name of the reagent</th>
			<th>Company</th>
			<th>Catalogue number</th>
			<th>Comments (optional)</th>
		</tr>
		<tr>
			<td>Green Food Dye</td>
			<td>McCormick</td>
			<td>n/a</td>
			<td>Must be filtered</td>
		</tr>
		<tr>
			<td>Hamilton Glass Syringe (100 &mu;L)</td>
			<td>Sigma Aldrick</td>
			<td>20702</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>LuerMxF Thread Style White Nylon</td>
			<td>Small Parts, Inc.</td>
			<td>VPLF-LC78-1-25</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Fine gauge Hypodermic Needles</td>
			<td>Popper</td>
			<td>7111</td>
			<td>Size: 33(SWG) x &frac14;" (6.35 MM)</td>
		</tr>
		<tr>
			<td>Wee Sight Transilluminator</td>
			<td>Respironics</td>
			<td>1017920</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>2.25X Headband Magnifier</td>
			<td>MagEyes</td>
			<td>Model No. 5</td>
			<td>Select magnification to fit individual preferences</td>
		</tr>
	</tbody>
</table>

delivery of therapeutic agents through intracerebroventricular (icv) and intravenous (iv) injection in mice

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most often results in failure of potential therapeutics designed for neurodegenerative disorders 1,2. Neurodegenerative diseases such as Spinal Muscular Atrophy (SMA), in which the lower motor neurons are affected, can benefit greatly from introducing the therapeutic agents into the CNS. The purpose of this video is to demonstrate two different injection paradigms to deliver therapeutic materials into neonatal mice soon after birth. One of these methods is injecting directly into cerebral lateral ventricles (Intracerebroventricular) which results in delivery of materials into the CNS through the cerebrospinal fluid 3,4. The second method is a temporal vein injection (intravenous) that can introduce different therapeutics into the circulatory system, leading to systemic delivery including the CNS 5. Widespread transduction of the CNS is achievable if an appropriate viral vector and viral serotype is utilized. Visualization and utilization of the temporal vein for injection is feasible up to postnatal day 6. However, if the delivered material is intended to reach the CNS, these injections should take place while the blood brain barrier is more permeable due to its immature status, preferably prior to postnatal day 2. The fully developed blood brain barrier greatly limits the effectiveness of intravenous delivery. Both delivery systems are simple and effective once the surgical aptitude is achieved. They do not require any extensive surgical devices and can be performed by a single person. However, these techniques are not without challenges. The small size of postnatal day 2 pups and the subsequent small target areas can make the injections difficult to perform and initially challenging to replicate.

Watch this Scientific Journal Video about Delivery of Therapeutic Agents Through Intracerebroventricular ICV and Intravenous IV Injection in Mice at JoVE.com

Delivery of Therapeutic Agents Through Intracerebroventricular ICV and Intravenous IV Injection in Mice

This article demonstrates two very different methods of injection: 1) into the brain (intracerebroventricular) and 2) systemic (intravenous) to introduce the therapeutic agents into the central nervous system of neonatal mice.

Delivery of Therapeutic Agents Through Intracerebroventricular (ICV) and Intravenous (IV) Injection in Mice

Despite the protective role that blood brain barrier plays in shielding the brain, it limits the access to the central nervous system (CNS) which most ...

delivery-therapeutic-agents-through-intracerebroventricular-icv

Research

JoVE Journal

Medicine

58.4K visualizações.  University of Missouri. Este artigo demonstra dois métodos muito diferentes de injecção: 1) para o cérebro (intracerebroventricular) e 2) sistêmica (intravenosa) para introduzir a agentes terapêuticos para o sistema nervoso central de camundongos recém-nascidos.