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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Murine Ommaya Assembly and Implantation
<ol>
	<li>Preparation of station
	<ol>
		<li>Disinfect the station surface. Place a blue covering over the surface and keep ready all sterilized tools and supplies indicated in&#160;Figure 1.</li>
	</ol>
	</li>
	<li>Presurgical procedure
	<ol>
		<li>Apply analgesia (Bup-SR)...

intracerebroventricular injection in mouse: an ommaya mediated direct drug administration method to deliver drugs to cerebral ventricles of mouse brain

Source: Law, V. et al. <a href="https://www.jove.com/v/62033/a-murine-ommaya-xenograft-model-to-study-direct-targeted-therapy">A Murine Ommaya Xenograft Model to Study Direct-Targeted Therapy of Leptomeningeal Disease.</a> J. Vis. Exp. (2021)
This video demonstrates ommaya mediated intracerebroventricular injection in a mouse model. The method enables localized delivery of potential drug candidates and evades the blood-brain barrier.

<img alt="Figure 1" src="/files/ftp_upload/20612/20612_Figure1.jpg" /> 
Figure 1: An example of a workstation setup for performing the Murine Ommaya implantation in mice.&#160;(1) Gas anesthesia machine/vaporizer. (2) Sterile blue paper drape covering a stereotaxic stand. (3) Stereotaxic device (stand/stage, ear bars, nose cone). (4) Microdrill. (5) Magnifying glass with light. (6) Sterile cotton-tipped applicator sticks with sterile saline rinse container. (7) Hydrogen peroxide. (8) Bead sterilizer.
<img alt="Fi...

Intracerebroventricular Injection in Mouse: An Ommaya Mediated Direct Drug Administration Method to Deliver Drugs to Cerebral Ventricles of Mouse Brain

Source: Law, V. et al. A Murine Ommaya Xenograft Model to Study Direct-Targeted Therapy of Leptomeningeal Disease. J. Vis. Exp. (2021)
This video ...

In leptomeningeal disease, cancer cells from other body organs metastasize to cerebrospinal fluid or CSF contained within leptomeninges - thin tissue layers surrounding the brain.
The intracerebroventricular injection facilitates direct drug administration in CSF, evading the blood-brain barrier.
To begin, prep an anesthetized mouse in the prone position.
Incise the head to reveal the skull. Locate the bregma - the junction point of coronal and sagittal sutures.
Using a microdrill, drill a hole in the skull at an appropriate distance from the bregma to access the outermost meningeal layer of dense connective tissue - the dura mater.
Then, gently insert the ommaya - a small injection port - into the hole.
Once done, suture the surrounding skin and allow the ommaya to affix for an optimum duration.
Next, connect the syringe filled with the desired drug suspension to the ommaya.
After setting a suitable flow rate, inject the drug suspension into the dura mater.
The ommaya provides a sterile pathway for direct drug delivery into the CSF in the cerebral ventricles of the brain.
Now, detach the syringe and leave the ommaya intact in the skull for subsequent dosage.
Allow the mouse to recover.
Over time, the drug molecules migrate towards the cancer site and prevent the growth and proliferation of cancer cells.

intracerebroventricular-injection-mouse-an-ommaya-mediated-direct

Research

JoVE Encyclopedia of Experiments

Cancer Research

Cancers of the Nervous System

In vivo studies

Intracerebroventricular Treatment with Resiniferatoxin and Pain Tests in Mice

The transient receptor potential vanilloid type 1 (TRPV1), a thermosensitive cation channel, is known to trigger pain in the peripheral nerves. In addition to its peripheral function, its involvement in brain functions has also been suggested. Resiniferatoxin (RTX), an ultrapotent TRPV1 agonist, has been known to induce long-term desensitization of TRPV1, and this desensitization has been an alternative approach for investigating the physiological relevance of TRPV1-expressing cells. Here we describe a protocol for intracerebroventricular (i.c.v.) treatment with RTX in mice. Procedures are described for testing nociception to peripheral TRPV1 stimulation (RTX test) and mechanical stimulation (tail pressure test) then follow. Although the nociceptive responses of mice that had been administered RTX i.c.v. were comparable to those of the control groups, RTX-i.c.v.-administered mice were insensitive to the analgesic effect of acetaminophen, suggesting that i.c.v. RTX treatment can induce supraspinal-selective TRPV1 desensitization. This mouse model can be used as a convenient experimental system for studying the role of TRPV1 in brain/supraspinal function. These techniques can also be applied to studies of the central actions of other drugs.

The transient receptor potential vanilloid type 1 (TRPV1) in the supraspinal region has been suggested to play some roles in the brain function. Described here is a protocol for intracerebroventricular injection of resiniferatoxin for supraspinal TRPV1 desensitization in mice. Procedures for some pain tests are also presented.

The transient receptor potential vanilloid type 1 (TRPV1), a thermosensitive cation channel, is known to trigger pain in the peripheral nerves. In ...

JoVE Journal

Neuroscience

Delivery of Antibodies into the Murine Brain via Convection-enhanced Delivery

Convection-enhanced delivery (CED) is a neurosurgical technique enabling effective perfusion of large brain volumes using a catheter system. Such an approach provides a safe delivery method by-passing the blood brain barrier (BBB), thus allowing treatment with therapeutics with poor BBB-permeability or those for which systemic exposure is not desired, e.g., due to toxicity. CED requires optimization of the catheter design, injection protocol, and properties of the infusate. With this protocol we describe how to perform CED of a solution containing up to 20 &#181;g of an antibody into the caudate putamen of mice. It describes preparation of step catheters, testing them in vitro and performing the CED in mice using a ramping injection program. The protocol can be readily adjusted for other infusion volumes and can be used for injecting various tracers or pharmacologically active or inactive substances, including chemotherapeutics, cytokines, viral particles, and liposomes.

Convection-enhanced delivery (CED) is a method enabling effective delivery of therapeutics into the brain by direct perfusion of large tissue volumes. The procedure requires the use of catheters and an optimized injection procedure. This protocol describes a methodology for CED of an antibody into a mouse brain.

Convection-enhanced delivery (CED) is a neurosurgical technique enabling effective perfusion of large brain volumes using a catheter system. Such an ...

Adaptable Angled Stereotactic Approach for Versatile Neuroscience Techniques

Stereotactic surgery is an essential tool in the modern neuroscience lab. However, the ability to precisely and accurately target difficult-to-reach brain regions still presents a challenge, particularly when targeting brain structures along the midline. These challenges include avoiding of the superior sagittal sinus and third ventricle and the ability to consistently target selective and discrete brain nuclei. In addition, more advanced neuroscience techniques (e.g., optogenetics, fiber photometry, and two-photon imaging) rely on targeted implantation of significant hardware to the brain, and spatial limitations are a common hindrance. Presented here is a modifiable protocol for stereotactic targeting of rodent brain structures using an angled coronal approach. It can be adapted to 1) mouse or rat models, 2) various neuroscience techniques, and 3) multiple brain regions. As a representative example, it includes the calculation of stereotactic coordinates for targeting of the mouse hypothalamic ventromedial nucleus (VMN) for an optogenetic inhibition experiment. This procedure begins with the bilateral microinjection of an adeno-associated virus (AAV) encoding a light-sensitive chloride channel (SwiChR++) to a Cre-dependent mouse model, followed by the angled bilateral implantation of fiberoptic cannulae. Using this approach, findings show that activation of a subset of VMN neurons is required for intact glucose counterregulatory responses to insulin-induced hypoglycemia.

Described here is a stereotactic procedure that can target challenging and difficult-to-reach brain regions (due to spatial limitations) using an angled coronal approach. This protocol is adaptable to both mouse and rat models and can be applied to diverse neuroscientific applications, including cannula implantation and microinjections of viral constructs.

Stereotactic surgery is an essential tool in the modern neuroscience lab. However, the ability to precisely and accurately target difficult-to-reach brain ...

Intracerebroventricular Injection in Mouse: An Ommaya Mediated Direct Drug Administration Method to Deliver Drugs to Cerebral Ventricles of Mouse Brain

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